Location: Temperate Tree Fruit and Vegetable Research
Project Number: 2092-22430-003-001-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Mar 31, 2019
End Date: Mar 30, 2021
Objective:
Identify invasive insect pests of tree fruits found in BC solid waste.
Quantify the risk of introduction of potential invasive insect pests of tree fruits from BC to WA through the solid waste transportation pathway.
Assess the establishment and spread risk of potential tree fruit invasive insect pests from BC to WA. Compare and match the climatic conditions of solid waste source sites with that of waste processing and receiving sites to assess the risk of tree fruit pest establishment. Sampling of green yard waste is underway in B.C. Shipment of samples will take place at regular intervals over the season. DNA extractions will be conducted over the winter months after field season. eDNA samples will be pooled and sent off for sequencing around February. Primer sets will be designed to amplify diagnostic regions of the COI for all the target species.
Approach:
We propose a collaborative research approach that involves personnel from the Temperate Tree Fruit and Vegetable Research Unit, USDA-ARS, Wapato, WA, and the Center for Sustainable Food Systems, University of British Columbia (UBC), Canada. Green yard waste sampling will be restricted to BC because the current issue raised by NWHC is green yard waste from BC as opposed to western WA; in addition, there will not be enough resources available through this proposal to conduct a more extensive study that includes surveying western WA, even though this would also be desirable. Surveying and modeling are the two major approaches selected in this study. Surveying will be conducted to sample green yard waste from the Vancouver area. Representative samples will be collected from six municipal yard waste and composting facilities in BC in 2018 and 2019. These sites were previously sampled by one of our cooperators in BC for nutrient and other analyses (Smukler et al. 2015; Zhang et al. 2017), but no invasive pest sampling has been conducted. Our BC cooperators will coordinate with these facilities to sample processed waste throughout the year (minimum of 4 sampling times/year), as the source materials vary seasonally. Samples will be pulverized, frozen, and transported to USDA-ARS laboratory in Wapato for pest identification using current protocols (see below for details). Internal (USDA-ARS, Wapato) and external specialists will use molecular barcoding techniques to identify the insect pests. Moreover, detailed information on the range and distribution of invasive species present in BC will be compiled from secondary sources.
Green yard waste samples (6 per site per time period) of 10 to 20 lbs will be collected and subjected to grinding to produce particles near sawdust size. Ground samples will be shaken to evenly distribute the particles in the sample. Sub-samples (5 per sample) of 100 g will be taken and stored at -20°C until it is needed for DNA extraction. Frozen samples will be shipped from Vancouver BC to Wapato for DNA extraction and amplification. A 6 g sample of each sub sample will be used for DNA extraction. The remaining sample will be stored at -80°C for future analyses or to serve as vouchers. The mannitol-PBS based CTAB DNA extraction method will be used (Fatima et al. 2014).
