Development of paratransgenic Artemia as a platform for control of infectious diseases in shrimp mariculture.
OBJECTIVE
To study the accumulation and retention of recombinant proteins in Artemia gut for optimizing paratransgenic disease control in shrimp aquaculture.


RESULTS
Transgenic Escherichia coli expressing fluorescent marker proteins and the transgenic cyanobacterium Synechococcus bacillarus expressing a functional murine single chain antibody, DB3, were fed to Artemia franciscana. Stable expression and retention of several marker molecules (e.g. GFP, DS Red and DB3) up to 10 h after of feeding with E. coli were evident within the gut of Artemia. Engineered strains of S. bacillarus expressing DB3 accumulated within the gut of Artemia with detectable antibody activity for 8-10 h of feeding via ELISA, coincident with the time period of the highest density of transgenic S. bacillarus in the Artemia gut.


CONCLUSIONS
Artemia fed transgenic bacteria or algae accumulated recombinant proteins for up to 10 h that retained biological activity. Co-delivery of multiple recombinant proteins simultaneously in the gut of Artemia was also demonstrated.


CONCLUSIONS
Expression of molecules that target infectious agents of mariculture in shrimp via commonly deployed feed organisms such as Artemia could potentially offer powerful new tools in the ongoing global effort to increase food supply.
Subhadra
B
B
 Department of Internal Medicine, School of Medicine, University of New Mexico, Albuquerque, NM, USA.
Hurwitz
I
I
 Department of Internal Medicine, School of Medicine, University of New Mexico, Albuquerque, NM, USA.
Fieck
A
A
 Department of Internal Medicine, School of Medicine, University of New Mexico, Albuquerque, NM, USA.
Rao
D V S
DVS
 Emeritus Scientist, Bedford Institute of Oceanography, Dartmouth, NS, Canada.
Subba Rao
G
G
 Shree Vasudha Laboratories, Visakhapatnam, India.
Durvasula
R
R
 Department of Internal Medicine, School of Medicine, University of New Mexico, Albuquerque, NM, USA.
D000818:Animals; D017756:Aquaculture / Q000379:methods*; D001156:Artemia / Q000382:microbiology*; D004926:Escherichia coli / Q000235:genetics*; D041981:Gastrointestinal Tract / Q000378:metabolism / Q000382:microbiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D030781:Organisms, Genetically Modified; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012758:Shellfish / Q000382:microbiology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000378:metabolism*; D046940:Synechococcus / Q000235:genetics*
D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D049452:Green Fluorescent Proteins
Blum-Oehler, G., Oswald, S., Eiteljorge, K., Sonnenborn, U., Schulze, J., Kruis, W. and Hacker, J. (2003) Development of strain-specific PCR reactions for the detection of the probiotic Escherichia coli strain Nissle 1917 in fecal samples. Res Microbiol 154, 59-66.
Cabello, F.C. (2006) Heavy use of prophylactic antibiotics in aquaculture: a growing problem for human and animal health and for the environment. Environ Microbiol 8, 1137-1144.
Chair, M., Nelis, H.J., Leger, P., Sorgeloos, P. and De Leenheer, A.P. (1996) Accumulation of trimethoprim, sulfamethoxazole, and N-acetylsulfamethoxazole in fish and shrimp fed medicated Artemia franciscana. Antimicrob Agents Chemother 40, 1649-1652.
Dante, K., Natividad, T., Hagio, M., Tanaka, M., Nomura, N. and Matsumura, M. (2007) White spot syndrome virus (WSSV) inactivation in Penaeus japonicus using purified monoclonal antibody targeting viral envelope protein. Aquaculture 269, 54-62.
Destoumieux, D., Munoz, M., Bulet, P. and Bachère, E. (2000) Penaeidins, a family of antimicrobial peptides from penaeid shrimp (Crustacea, Decapoda). Cell Mol Life Sci 57, 1260-1271.
Dhont, J. and Sorgeloos, P. (2002) Applications of Artemia. In Artemia: Basic and Applied Biology ed. Abatzopoulos, T.J., Beardmore, J.A., Clegg, J.S. and Sorgeloos, P. pp. 251-272. the Netherlands: Kluwer Academic Publications.
Durvasula, R.V., Gumbs, A., Panackal, A., Kruglov, O., Aksoy, S., Merrifield, R.B., Richards, F. and Beard, C.B. (1997) Prevention of insect-borne disease: an approach using transgenic symbiotic bacteria. Proc Natl Acad Sci USA 94, 3274-3278.
Durvasula, R.V., Gumbs, A., Panackal, A., Whitham, R., Taneja, J., Kang, A., Richards, F. and Beard, C.B. (1999) Expression of a functional antibody fragment in the gut of Rhodnius prolixus via the transgenic Bacterial Symbiont, Rhodococcus rhodnii. Med Vet Entomol 13, 1-5.
Durvasula, R.V., Sundaram, R., Matthews, S., Sundaram, P. and Subba Rao, D.V. (2006) Prospects of paratransgenic applications to commercial mariculture using genetically engineered algae. In Algal Cultures, Analogues of Blooms and Applications ed. Subba Rao, D.V. pp. 865-889. Enfield, NH, USA: Science Publishers.
Durvasula, R., Hurwitz, I., Subhadra, B. and Matthews, S. (2007) Regulatory issues with biotechnology; International Congress of Insect Biotechnology & Industry, 19-24 August, Daegu, Republic of Korea. Entomol Res 37(Suppl 1), A69-A73.
Durvasula, R.V., Subhadra, B., Hurwitz, I. and Subba Rao, D.V. (2009) Paratransgenic approaches to the control of infections of shrimp: prospects for use of Dunaliella. In The Alga Dunaliella: Biodiversity, Physiology, Genomics and Biotechnology ed. Ben-Amotz, A., Polle, J.E.W. and Subba Rao, D.V. pp. 385-402. NH, USA: Science Publishers.
FAO (2007) The State of World Fisheries and Aquaculture 2007. Rome, Italy: Food and Agriculture Organization Publications.
Grozdanov, L., Raasch, C., Schulze, J., Sonnenborn, U., Gottschalk, G., Hacker, J. and Dobrindt, U. (2004) Analysis of the genome structure of the nonpathogenic probiotic Escherichia coli strain Nissle 1917. J Bacteriol 186, 5432-5441.
Hill, B.J. (2005) The need for effective disease control in international aquaculture. In Progress in Fish Vaccinology, Developments in Biologicals 121 ed. Midtlyng, P.J. pp. 3-12. Basel, Switzerland: Karger Publications.
Hillesland, H., Read, A., Subhadra, B., Ghosh, K., Das, P., Hurwitz, I. and Durvasula, R. (2008) Identification of aerobic gut bacteria from the Kala Azar Vector, Phlebotomus argentipes: a platform for potential paratransgenic manipulation of sandflies. Am J Trop Med Hyg 79, 881-886.
Johnson, K.N., Van Hulten, M.C.W. and Barnes, A.C. (2008) Vaccination of shrimp against viral pathogens: phenomenology and underlying mechanisms. Vaccine 26, 4885-4892.
Luo, T., Li, F., Lei, K. and Xun, X. (2007) Genomic organization, promoter characterization and expression profiles of an antiviral gene PmAV from the shrimp Penaeus monodon. Mol Immunol 44, 1516-1523.
Marques, A., Dhont, J., Sorgeloos, P. and Bossier, P. (2004) Evaluation of different yeast cell wall mutants and microalgae strains as feed for gnotobiotically-grown brine shrimp Artemia franciscana. J Exp Mar Biol Ecol 312, 115-136.
Miller, T.A., Lauzon, C.R., Lampe, D., Durvasula, R. and Matthews, S. (2006) Paratransgenesis applied to control insect-transmitter plant pathogens: the Pierce’s disease case. In Insect Symbiosis, 2nd edn ed. Bourtzis, K. and Miller, T.A. pp. 243-267. Boca Raton, FL, USA: CRC Press, Taylor and Frances.
Robalino, J., Bartlett, T., Shepard, E., Prior, S., Jaramillo, G., Scura, E., Chapman, R.W., Gross, P.S. et al. (2005) Double-stranded RNA induces sequence-specific antiviral silencing in addition to nonspecific immunity in a marine shrimp: convergence of RNA interference and innate immunity in the invertebrate antiviral response? J Virol 79, 13561-13571.
Robalino, J., Bartlett, T., Shepard, E., Prior, S., Jaramillo, G., Scura, E., Chapman, R.W., Gross, P.S. et al. (2007) Double-stranded RNA and antiviral immunity in marine shrimp: inducible host mechanisms and evidence for the evolution of viral counter-responses. Dev Comp Immunol 31, 539-547.
Sundaram, R.K., Hurwitz, I., Matthews, S., Hoy, E., Kurapati, S., Crawford, C., Sundaram, P. and Durvasula, R.V. (2008) Expression of a functional single-chain antibody via Corynebacterium pseudodiphtheriticum. Eur J Clin Microbiol Infect Dis 27, 617-622.
Witteveldt, J., Cifuentes, C.C., Vlak, J.M. and Van Hulten, M.C. (2004) Protection of Penaeus monodon against white spot syndrome virus by oral vaccination. J Virol 78, 2057-2066.
Yi, G., Wang, Z., Yao, L., Qian, J. and Hu, L. (2004) Vp28 of shrimp white spot syndrome virus is involved in the attachment and penetration into shrimp cell. J Biochem Mol Biol 30, 726-734.
Zasloff, M. (2002) Antimicrobial peptides of multicellular organisms. Nature 415, 389-395.