Performance Report for Cooperative Agreement No: na06oar4810163 for the Period from September 1, 2006 to August 31, 2012 University of Maryland Eastern Shore

Rash, J. M., D. McIntosh, D. Kapareiko, G. H. Wikfors, E. Schott, and H. J. Schreier. Biochemical and molecular analysis of bacteria used in protecting oyster larvae from pathogenic bacteria in hatchery culture. Fifth NOAA Center for Atmospheric Sciences (NCAS) Education and Science Forum, submitted.

Tagged unparasitized grass shrimp growth (in length) was higher than tagged parasitized grass shrimp for all 3 mesocosms. Tagged unparasitized grass shrimp grew an average (±SD) of 19.02±14.17% in length and tagged parasitized grass shrimp lost an average of -0.56±12.79% in length. There was significant difference in growth in length (mm) between tagged unparasitized and tagged parasitized grass shrimp (p<0.0001). Tagged unparasitized grass shrimp gained an average (±SD) of 82.57±78.20% in weight. Tagged parasitized grass shrimp gained an average (±SD) of 2.71±43.45% in weight. There was a significant difference in weight (g) change between tagged unparasitized and tagged parasitized grass shrimp (p<0.0001). There were no significant differences in initial lengths (p=0.1746) or initial weights (p=0.5354) of untagged unparasitized grass shrimp and tagged unparasitized grass shrimp. There were also no significant differences in final lengths (p=0.1006) or final weights (p=0.05) of unparasitized grass shrimp and tagged unparasitized grass shrimp. Thus, the tag does not affect growth of grass shrimp.

For the predation study, trial 1 (of 3) was excluded due to the escape of some of the predators (mummichog Fundulus heteroclitus). The mummichog ate more tagged parasitized shrimp than any other treatment. We found that unparasitized grass shrimp average (±SD) percent survival was the highest (41.00±14.38%) while tagged parasitized grass shrimp average (±SD) percent survival was the lowest (17.00±5.03). There was a significant difference (p=0.0199) in survival of grass shrimp type when the control mesocosm of each trial was included. Further analysis revealed that unparasitized grass shrimp survival was significantly different than tagged parasitized grass shrimp survival (p=0.0388). There was no significant difference in survival between the other types of grass shrimp.

Undergraduate student Joe LaBarre conducted a small study to determine whether parasitized grass shrimp are more susceptible to predation by the mummichog Fundulus heteroclitis. He has found that shrimp behavior is more important than whether the shrimp was parasitized by Propopyrus pandalicola, but has found that mummichog preferred parasitize shrimp 59% of the time. Undergraduate student Sheena Corning conducted a small project on the effect of the parasite on starvation in grass shrimp and found that parasitized shrimp survived 2-4 days less than unparasitized shrimp. Undergraduate student Michele Sherman is conducting a study to determine the effect of removing this parasite from the shrimp and has seen an intermediate survivorship rate.

Monitoring study for shrimp densities in Georgia is continuing. To date, the highest egg loss due to sterilization was 335 eggs/m³. In a separate study, mortality was higher among non-gravid female P. pugio (32%) than gravid P. pugio (24%) when exposed to fipronil for 96 h. Parasitic sterilization, causing a loss of 21% of mean egg production, and exposure to fipronil may reduce the number of eggs available by as much as 45%, which may lead to a decline of this important prey item. The results of this project include: a M.S. thesis in May 2009 entitled “Effect of two potential stressors on the grass shrimp Palaemonetes pugio in southeastern Georgia;” another M.S. thesis anticipated in December 2009 entitled: “The effects of coded wire tags and the isopod parasite Probopyrus pandalicola on the growth and predation of grass shrimp Palaemonetes pugio”; and a third thesis is anticipated in May 2010 entitled “Behavioral effects of the parasite Probopyrus pandalicola on the swimming endurance and toxicity of fipronil to the grass shrimp Palaemonetes pugio. A publication describing our previous year’s research is in press and due out this fall entitled “Toxicity of synthetic pyrethroid insecticides to the grass shrimp, Palaemonetes pugio, parasitized with the bopyrid isopod, Probopyrus pandalicola.” This is the first publication in Dr. Curran’s career that was coauthored with an undergraduate student. A second grass shrimp K-12 activity is also in press this year entitled “Bringing scientific inquiry alive using real grass shrimp research. This was coauthored with a previous teacher-intern funded by LMRCSC and she is now one of the teachers who invites SSU faculty and students to come to her class and speak about research. Three other K-12 activities are in review and are coauthored by the teacher-intern from 2008-2009. All the above participants have been funded by LMRCSC or have received field assistance and use of supplies through LMRCSC funding.

The success of the year I efforts has encouraged Dr. Curran to pursue grass shrimp research as the major focus of her research efforts. Therefore, she has used the LMRCSC preliminary data as the background for her PI/coPI roles in the following 3 funded proposals: Collaborative: NSF New GK12: Building Ocean Literacy in a Coastal Community through Science Education and Estuarine Monitoring; NSF OEDG Collaborative Research: Enhancing Diversity in Geoscience Education through Coastal Research in a Port City (EDGE); and Department of Education Title VII Coastal Ocean and Underwater Research to Advance Graduate Education (COURAGE).

Locus		Primer sequences
MF12		F: 5'-GACTGCTTCACCCACAGGAG-3'
		R: 5'-TTCAGATTTTGTTTTCCTCAAGG-3'
MF13		F: 5'-TAGGGCAGGGTGTGAACTTTAC-3'
		R: 5'-CTCTGCTGAGACCGTGACATAG-3'
MF14		F: 5'-GAAAAATTGGACAGATTGAGAGAG-3'
		R: 5'-CTTTAAGTTGATGCAGAAAATTCC-3'
MF16		F: 5'-GCTGGTGAGGTTTTCAAAGTG-3'
		R: 5'-AACACCATCCTCGCATATTCAC-3'
MF18		F: 5'-AGGGCAAGAGGTTAATAAGAGG-3'
		R: 5'-TTAGTGTTTCAGGGTCTTGCTC-3'
MF21		F: 5'-TCTTTCAACGACAGGACAGGA-3'
		R: 5'-AGAGGGGATGCTAATCTGCA-3'
MF27		F: 5'-CAAATGCCATTGGTTGAATTG-3'
		R: 5'-TTTCAAGTCATTGCATGTAGCAG-3'
MF28		F: 5'-TGCTGAATTGCTGCTGTTATT-3'
		R: 5'-GTGAACGTCTTTCTCCTCCAAC-3'

Monkfish samples will be collected from four nearshore (<50m depth) and four offshore (>100m depth) habitats in each of the management areas (Fig. 5). Because of the likelihood of long-distance movements of monkfish (Richards et al., 2008), all samples in the eight locations were collected within a two-week window. The samples were collected in 2009 by the Northeast Fisheries Science Center spring survey and the Cooperative Monkfish Survey (A. Richards, Chief Scientist). The Monkfish Survey cruises involved two charter vessels, one sampling in the NMA and another in the SMA. Additional samples were collected inshore from monkfish gillnetters participating in the Monkfish Set-Aside Program (2008/2009). Muscle and liver tissues from at least 50 fish per site were collected and frozen at -80C prior to processing. Whole genomic DNA was isolated using kits following manufacturer’s instructions (Zymo Research Corporation, Orange CA).

Three genetic groups were observed which consisted of L.a. I, L.a. II and L.a III. The non-spatial correlation of the groups suggests that the examined monkfish population may not be a unit stock. Genetic assortment does not correspond to either management area, implying that the monkfish in US waters are not geographically isolated. Two outliers were observed in the phylogenetic tree which may be an indication of the presence of another species of monkfish (Lophius gastrophysus) that is primarily distributed south of Cape Hatteras, NC. This will be verified in subsequent studies. PCR reliability with universal eukaryotic primers will also be checked to determine if the eukaryotic primer results would be consistent with the COX-I tested samples in this study.