2024
Continued identification and culturing of slug parasitic nematodes in Delaware
Contributor/Checkoff:
Category:
Sustainable Production
Keywords:
Cover cropsCrop protectionField management NematodePest
Lead Principal Investigator:
Michael Crossley, University of Delaware
Co-Principal Investigators:
Project Code:
24A00840
Contributing Organization (Checkoff):
Institution Funded:
Brief Project Summary:
In 2022, Delaware farms planted 160,000 acres of soybean, averaging 43 bu/acre with a value > $92 million (USDA NASS 2022). Slugs represent a severe, albeit sporadic pest of soybean, for which the costs of treating or not treating can be equally costly. Often, by the time stand loss is evident, the window of opportunity for rescue treatment is long gone. Knowledge about the natural enemy complex that interacts with slugs and of the factors that promote natural suppression of slugs is needed to support an integrated management approach for slugs in lowtill soybean.
Key Beneficiaries:
#farmers
Unique Keywords:
#parasitic nematode, #slug
Information And Results
Project Summary

Slugs are a persistent threat to Delaware soybean, typically infesting < 20% of soybean acreage but causing significant yield loss when populations reach high densities (Musser et al. 2018, 2019). The sporadic-but-severe nature of slug damage makes management frustrating. Ironically, insecticides make slug problems worse by killing predators but leaving slugs unharmed (Douglas et al. 2015). Molluscicides (e.g., metaldehyde or iron phosphate, applied as a bait) can be effective, but are too costly and prone to washing away with rain to be relied upon as a preventative treatment (Bailey 2002). By the time slug damage is evident, though, it may already be too late to achieve control with a molluscicide. This is a classic “damned if you do, damned if
you don’t” conundrum. Moving forward, we need to understand what factors put a soybean field at greater risk of economic damage by slugs so that we can manage our farms to avoid situations where stand loss becomes unacceptably high.

Natural predators and parasites (enemies) of slugs are a perhaps underappreciated ally in our battle against slugs. A variety of ground beetles, spiders, marsh flies, and nematodes are known to consume slugs at different parts of the slug life cycle (Barker 2004). These natural enemies are themselves influenced by a number of factors, including weather, tillage, pesticide use, and cover crops (Everts et al. 1989; Vernavá et al. 2004; Le Gall & Tooker 2017; Rivers et al. 2018). Sometimes, the link between natural enemy numbers and slug numbers appears clear. For example, a common ground beetle (Pterostichus melinarius) readily consumed a common slug (Deroceras reticulatum) in a small grains farm in the UK, and slug numbers dropped with increasing beetle numbers (Symondson et al. 2002). However, it is unclear how farms can leverage these natural enemies.

The Delaware Soybean Board provided support for Dr. Crossley shortly after his arrival at University of Delaware to begin examining the natural enemies that could make a dent in slug populations and the factors that promote these natural enemies. In 2023, Dr. Crossley, along with help from Dr. David Owens and the effort of a dedicated PhD student (Thabu Mugala), regularly sampled a total of 17 fields, yielding a total of 1,531 slugs (1,043 marsh slugs, 488 gray garden slugs). Only 20 marsh slugs (~2%) melted and produced nematodes, a low but typical proportion (~3% of slugs melted in 2022). These slugs originated in just two sites. No gray garden slugs were infected with nematodes. Identification is ongoing, but so far we have identified three of the nematode species. One of them, Panagrolaimus detritophagus (these critters don’t have common names, sorry), is not considered a strict parasite, but instead uses slug hosts to disperse and feeds on bacteria in the host and environment. The other, Pristionchus pacificus, is an obscure species that has only been recorded parasitizing scarab beetles. Finding it in a slug is exciting, but warrants further investigation to determine pathogenicity. The third species was identified to the genus Oscheius, which includes species that are known slug parasites. We are most excited about finding this nematode. We plan to continue identifying these nematodes, and to eventually conduct pathogenicity tests to verify the potential of these nematodes to serve as effective slug parasites.

Project Objectives

1) Continue to collect and identify slug parasitic nematodes from Delaware soybean fields.
2) Continue to develop a liquid culturing technique to maintain slug parasitic nematode colonies in the lab.

Project Deliverables

Objective 1: We will continue sampling slugs and natural enemies using shingle traps and pitfall traps placed within no-till soybean fields in the Spring of 2024 to collect and identify slug parasitic nematodes. Importantly, we are always looking for collaborating farms that are willing to let us sample slugs and natural enemies from their farms. Please reach out to Mike if interested (crossley@udel.edu). To collect slug parasitic nematodes, all captured slugs will be reared in the lab under ideal conditions for 3 weeks, at which point infected slugs will melt and nematodes will emerge seeking new hosts. Melted slugs will be placed in white traps (basically a petri dish with water to attract dispersing nematodes). A subsample of nematodes will be used for morphological identification and DNA sequencing (using the COI barcode) to confirm species identity.

Objective 2: We ultimately aim to test the pathogenicity of any slug parasitic nematodes that we collect, with the goal of developing a highly effective strain of nematodes that can be used for slug control. Work on this front is ongoing. Nematodes are currently being maintained in colonies that are occasionally replenished by infecting live slugs and collecting emergent
nematodes. However, to be able to determine nematode pathogenicity against slugs, we need to be able to maintain nematodes without slugs and ramp up their numbers in liquid culture. To do so, we have begun isolating bacteria from slugs and culturing them on agar petri dishes. The next step is to identify these bacteria, and then get them into liquid culture to feed nematode colonies. Toward this end, we just acquired an incubated orbital shaker, a necessary piece of equipment to
maintain these bacterial cultures (they need to be kept cool and aerated by continuous shaking). This equipment was purchased using my own lab startup funds.

Progress Of Work

Updated July 22, 2024:
Objective 1:
We collected 363 gray garden slugs and 272 marsh slugs from 13 fields in Delaware and Maryland between April and June. 30 gray garden slugs (8%) and 58 marsh slugs (21%) were infected with slug-parasitic nematodes. Identification of these nematodes to species using morphological and DNA sequence characteristics is underway.

Objective 2:
We have developed a workflow to maintain slug-parasitic nematodes in a lab culture. This entails culturing bacteria that the nematodes feed on in nutrient-enriched agar plates, then adding nematodes and allowing them to grow and reproduce for a period of time before transferring a sample of nematodes to a new bacterial colony to continue feeding, growing, and reproducing. This workflow will be used to generate enough slug-parasitic nematodes from each field collection to conduct pathogenicity experiments to identify slug-parasitic nematode species and strains that are highly pathogenic toward slugs and that could be promoted for commercial production.

Final Project Results

Benefit To Soybean Farmers

The United Soybean Research Retention policy will display final reports with the project once completed but working files will be purged after three years. And financial information after seven years. All pertinent information is in the final report or if you want more information, please contact the project lead at your state soybean organization or principal investigator listed on the project.