Megan Herlihy 1 , Elijah Talamas 2 and Donald Weber 1 1 USDA ARS , Beltsville, MD;

1. Parasitoids of Brown Marmorated Stink Bug (Halyomorphahalys) eggs: Comparison of three egg mass types in three Maryland habitats. Megan Herlihy1, Elijah Talamas2 and Donald Weber1 1USDA ARS, Beltsville, MD; 2USDA ARS, Washington, DC,

2. Introduction • BMSB invasive pest from Asia. • Many sentinel egg mass studies done, generally reporting low rates of parasitism by native parasitoids. • Potential biocontrol agent parasitoids (family Scelionidae) imported from Asia now in quarantine for non-target testing.

3. Methods: 3 Habitat types • Vegetable crop (soybean) • Orchard (apple) • Woods near stream (various native and invasive arboreal and herbaceous vegetation)

4. Methods:3 Egg mass Treatments • ≤24-hour old eggs laid by colony insects • ≤24-hour old eggs laid by colony insects then frozen at -80°C for 2 min. • Screen cages (containing 20 BMSB and 1 bean plant for oviposition) • Provides the host finding cues needed by parasitoids • Cages are less labor intensive/ time consuming than searching for field laid eggs as in Jones et al. (2014)

5. MethodsExperimental design • Fresh and frozen sentinel eggs laid on paper towels by colony insects were pinned to various vegetation at each site and were exposed for 72 hrs. • Cage laid eggs were collected twice weekly. (~ every 72hrs.) • All egg masses returned to lab and reared out in a growth chamber (16L:8D, 25°C)until either a BMSB nymph or a parasitoid emerged • If nothing emerged, eggs were dissected. • Emerged and dissected parasitoids were sent to Elijah Talamas (USDA ARS SEL) for identification

6. Methods:BMSB eggs exposed by habitat and egg mass type Total number of BMSB egg masses (# of individual BMSB eggs)

7. Hypothesis 1 • Frozen egg masses will result in higher rates of successful parasitism than fresh or cage egg masses at all sites, likely due to the lack of immune defenses of the egg. (we got this idea from Tim Haye of CABI in Switzerland)

8. Hypothesis 2 • Cage laid egg masses should result in higher parasitism than 24-hour colony laid sentinel egg masses, because it provides the parasitoids with host finding cues.

9. Hypothesis 3 • Inclusion of wooded habitat allowed us to find the exotic Trissolcus japonicus, previously thought to only be found in quarantine in the United States. • Trissolcus japonicus is the most successful parasitoid to complete development and emerge from live egg masses.

10. Hypothesis 4 • Parasitoid species are habitat specific.

11. Hypothesis 5 • Native parasitoids are only successfully developing and emerging from dead eggs.

12. Hypothesis 6 • There is a seasonal pattern to the occurence of parasitoids as a group or as individual species.

13. Results:H°1 Frozen egg masses will result in higher parasitism rates Yes! • Overall, frozen egg masses (25%)more likely to yield adult parasitoids versus fresh (7%). (Fisher P1-tail p=0.000126) • Parasitoids more likely to develop to adulthood from frozen (16%), versus fresh (4%) host eggs ( χ²=253Pearson p<0.0001)   • Parasitoids more likely to successfully emerge from frozen (10%), versus fresh host eggs (3%) ( χ²=101.2Pearson p<0.0001)

14. Recovered parasitoid key

15. Results:H2: Cage laid egg masses should result in higher parasitism Because we used bean plants in all cages, we only saw Telenomus podisi in the cage treatments

16. Results:H3: Trissolcusjaponicusis the most successful parasitoid • Yes! • T. japoncisis much more successful at emerging from fresh egg masses than native parasitoids. (Fishers exact p<0.001) • Native parasitoids are more successful at emerging from frozen egg masses than T. japonicus.(Fishers exact p<0.001)

17. Results:H4: Parasitoids are habitat specific • Yes! • T. japonicusand T. brochymenaefound only in the woods. • T. podisifound only in the soybean field and in the cages with bush beans. • Anastatussp. found only in the orchard. • T. euschistifound in both the woods and the orchard.

18. Results:H5: Native parasitoids are only successfully developing and emerging from dead eggs. • Can’t prove this yet, but… 19% of control eggs never emerged. • This easily accounts for the 7% of egg masses successfully parasitized by natives.

19. Results:H6: There is a seasonal pattern to the occurrence of parasitoids. • Not enough data to tell.

20. Future research • Expand the geographical range where we place sentinels and cages to define the distribution of T. japonicus. • Improve the cage treatment: Use sleeve cages on existing vegetation in place of bean plants.  • Include native beneficial hemipteran egg masses in future sentinel studies (e.g. Podisus maculiventris)

21. 2015 plan for sentinel egg mass detection of • Trissolcusjaponicus and other parasitiods • Nearby sites BARC West: every week • North Farm original site (woods) • + 4 additional woody sites • 4 sites of orchard/savannah habitat • 4 sites of field crop (soybean) habitat • Satellite wooded sites: every four weeks, total 4 times during season • BARC South Farm (North area) • Linkage Farm (between RI Avenue and Metro yard) • Little Paint Branch North (north of Sellman Avenue) • Beaver Dam Road (BARC East, west of BW Parkway) • Collaborator sites: • College Park, BARC East of BW Parkway, Port of Baltimore (UMd.) • Newark Delaware vicinity and other sites (ARS BIRL) • Area of Harpers Ferry & Northern Shenandoah (ARS AFRL & Va. Tech) BMSB reared, frozen, and Podisus egg masses (3 plant hosts X 2 e.m.) frozen BMSB egg masses only (3 plant hosts)

22. Acknowledgements • Abby Rosenberg, Nate Erwin, Gabriel York and Emma Thrift for their field work and other help!