Progress 07/15/06 to 06/14/08
Outputs
OUTPUTS: We developed a microarray for Litopenaeus vannamei from 156,952 EST sequences from NCBI. This resulted in 12,918 "sense" probes for 5,137 transcripts, and 12,897 "antisense" probes for 5,141 transcripts. The final shrimp array contains 25,815 unique probes for 5,499 transcripts. We evaluated grass shrimp and blue crab. The virus n grass shrimp can replicate for 36d and reached a peak on 14d. The mortality is 8% by injection. Grass shrimp showed light infection after feeding YHV-positive L. vannamei during 21d period. The mortality is 0 % by feeding. L. vannamei can be infected by YHV after feeding YHV-positive grass shrimp. Grass shrimp is a reservoir host for YHV. Grass shrimp is a good animal model in YHV research. For the blue crab, When the blue crab was injected with YHV, the viral load decreased rapidly after 3 dpi. When the blue crab was fed with YHV, the viral load increased and peaked on 3 dpi and then decreased after 7 dpi. The blue crab is a short term carrier (vector) host for YHV. We evaluated the effect of temperature on TSV epidemics and showed experimentally that the basic reproduction number was negatively correlated with temperature. We examined the with-in host genetic variability of TSV infections and compared acutely infected shrimp to chronically infected. The PCR targeted CP2 (VP1) region (1303 bp) of the Tx95 with high fidelity pfu polymerase. We produced 130 Acute clones from hemolymph of 5 shrimp/ Average of 21.6 clones per shrimp/ Sequenced product of each clone. In acute hemolymph, of 130 Acutes: 113 are identical to the one from the 94USHI1 isolate. Sixteen clones showed 1 nucleotide change, 1 clone had 2 nucleotide changes (Nucleotide 516 and 661 were concurrently mutated, same mutation appeared in 3 chronics). Quasispecies complexity (Pn) as calculated by # of polymorphic sites/# nucleotides sequenced in Acutes = 1.18 x 10^-4. The Normalized Shannon diversity = 0.1167. For chronically infected shrimp we produced 94 clones from 5 chronic shrimp hemolymph, 18.8 clones per shrimp. In Chronic Hemolymph: 75 are identical to the one from the 94USHI1. 14 clones showed 1 nucleotide change. Of the 14, 4 from 3 different shrimp had the change from C to T at position 1037. Four clones had 2 nucleotide changes (Position 516 and 661 were again concurrently mutated in a sequences - same changes). One clone had 3 nucleotide changes. Quasispecies complexity (Pn) as calculated by # of polymorphic sites/# nucleotides sequenced in Chronic Hemolymph: Pn = 2.04 x 10^-4. Normalized Shannon Entropy (Sn) = 0.189. We evaluated the effect of floc inoculations on production of shrimp in a large-scale commercial prototype RAS for shrimp. With 10% volume of floc inoculation we produced 4.4 kg of shrimp per cubic meter. Adding no inoculation we achieved only 3.0 kg of shrimp per cubic meter. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The development of the microarray will allow us to elucidate the mechanisms of shrimp TSV resistance. The identification of reservoir hosts of YHV allows farmers to better develop biosecurity protocols to prevent YHV outbreaks. The within host analysis of TSV genetic diversity will better allow us to understand the evolution and pathogenesis of TSV. The floc experiment allows farmers to better manage the water quality of recirculating aquaculture systems for shrimp production.
Publications
- Ma, H., R.M. Overstreet, J. A. Jovonovich. 2008. Stable yellowhead virus (YHV) RNA detection by qRT-PCR during six-day storage. Aquaculture 278: 10 - 13.
- Ogle, J.T. A.V. Flosenzier and J.M. Lotz. 2006. USM-GCRL Large Scale Growout Marine Shrimp Production Facility. In Sea Grant Publication VSG-06-05. Proceedings The Sixth International Conference on Recirculating Aquaculture. July 21-23 2006. p6 - 13.
- Lotz, Jeffrey M., Robin M. Overstreet, and D. Jay Grimes. 2006. Aquaculture and animal pathogens in the marine environment with emphasis on marine shrimp viruses. In Oceans and Health: Pathogens in the Marine Environment, edited by Shimshon Belkin and Rita R. Colwell. Springer (Kluwer Academic/Plenum Publishers), New York, pp. 431 - 451.
- Shields, Jeffrey D. and Robin M. Overstreet. 2007. Parasites, diseases, and symbionts. In V.S. Kennedy and L. E. Cronin (eds). The Blue Crab, Callinectes sapidus. Maryland Sea Grant.
- Vincent, A.G. and J.M. Lotz. 2007. Advances in research of necrotizing hepatopancreatitis bacterium (NHPB) affecting penaeid shrimp aquaculture. Reviews in Fisheries Science 15: 63 - 73.
- Ogle, JT & JM Lotz. 2006 effect of different greenhouse covers on production of white shrimp Litopenaeus vannamei in a pilot scale facility. Aquaculture America 2006. Annual Meeting of the US Chapter of the World Aquaculture Society. Las Vegas, Nevada.
- Vincent, A.G. and J.M. Lotz. 2007. Effect of salinity on transmission of necrotizing hepatopancreatitis bacterium (NHPB) to Kona stock Litopenaeus vannamei. Diseases of Aquatic Organisms 75: 265 - 268.
- Lotz, J.M. 2007. Disease Control Strategies for Shrimp Farming. Aquaculture 2007. Annual international meeting of the World Aquaculture Society. San Antonio, Texas, USA.
- Lotz, J.M. and J.T. Ogle. 2007. Overview of The University of Southern Mississippi - Gulf Coast Research Lab large - scale growout facility. Annual international meeting of the World Aquaculture Society. San Antonio, Texas, USA.
- Lotz, JM & AG Vincent. 2006.advances in research on Necrotizing Hepatopancreatitis (NHP). Aquaculture America 2006. Annual Meeting of the US Chapter of the World Aquaculture Society. Las Vegas, Nevada.
- Zimmerman, NP, V. Breland and J.M. Lotz. 2006. Regulation of Taura syndrome virus by shrimp that survive initial infection. Aquaculture America 2006. Annual Meeting of the US Chapter of the World Aquaculture Society. Las Vegas, Nevada.
- Ma, Hongwei, Ph.D. 2008. Yellowhead virus: transmission and genome analyses. in crustacean hosts. The University of Southern Mississippi. Vincent, Amanda G. 2006. Experimental infection of necrotizing hepatopancreatitis bacterium (NHPB) in Kona stock Litopenaeus vannamei (Pacific white shrimp). Ph.D. Dissertation. The University of Southern Mississippi.
- Ostrowski, A.C., H. Atwood, A. Alcivar-Warren, Q.C. Fontenot, A.L. Lawrence, D.V. Lightner, J.M. Lotz and S.M. Moss. 2007. Milestone achievements and future directions of the U.S. Marine Shrimp Farming Program in response to evolving world markets. . Annual international meeting of the World Aquaculture Society. San Antonio, Texas, USA.
- Ostrowski, AC, HL Atwood, A. Alcivar-Warren, Q Fontenot, AL Lawrence, DV Lightner, JM Lotz & SM Moss. 2006 Recent progress and milestone achievements of the US Marine Shrimp Farming Program in response to evolving world markets. Aquaculture America 2006. Annual Meeting of the US Chapter of the World Aquaculture Society. Las Vegas, Nevada.
- Vincent. A. G. and J. M. Lotz. 2006. A model of Necrotizing Hepatopancreatitis-Bacterium NHP-B) epidemics in its shrimp host Litopenaeus vannamei. Aquaculture America 2006. Annual Meeting of the US Chapter of the World Aquaculture Society. Las Vegas, Nevada.
- Vincent, Amanda G. and J. M. Lotz. 2006. Evolution of virulence: transmission from dead hosts. Southeastern Society of Parasitologists, joint meeting with the Association of Southeastern Biologists. Gatlinburg, TN.
- Tremonte, Jocelyn. 2008. The effects of temperature on Taura syndrome virus (TSV) infections in Litopenaeus vannamei. The University of Southern Mississippi.
- De la Calzada, Rey. 2008. Comparison of White Spot Syndrome Virus LD50s in Litopenaeus vannamei and Farfantepenaeus duorarum. The University of Southern Mississippi.
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