Progress 07/01/11 to 09/30/15
Outputs
Target Audience:The global (Latin American, SE Asia, South Asian, East African, and Middle Eastern) shrimp farming industries. Changes/Problems:Most of our work was completed in 2013, therefore, 2014 and 2015 work were focused on an emerging disease, AHPND. What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?Yes and the PCR methods detecting AHPND-V. parahaemolyticus are published and have been distributed to interested countries including China, Vietnam, Thailand, Malaysia, Indonesia and Mexico where EMS/AHPND could become a major disease. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Since last period, our research has been shifted to a bacterial disease: AHPND (also referred as early mortality syndrome). In 2014, we have determined that V. parahaemolyticus is the cause of this disease. In this period, we identified a virulent plasmid (69-kb, named as pVPA3-1) that encodes proteins similar to Photorhabdus insect-related (Pir) toxins. Based on a proteomic analysis, the pirA-like (336 bp) and pirB-like (1317 bp) genes encode for 13 and 50 kDa proteins, respectively; and these two proteins were determined to the toxins responsible for AHPND. Subsequently, we developed a duplex PCR diagnostic method, which can reliably detect AHPND-causing strains of V. parahaemolyticus and does not cross react with non-pathogenic strains or with other species of Vibrio bacteria. Furthermore, we developed a qPCR assay targeting the pirA-like gene to detect and quantify the virulence plasmid harbored by the AHPND-V. parahaemolyticus. This assay is specific with high sensitivity (10 copies of virulence plasmid) and can be used to detect AHPND-V. parahaemolyticus in shrimp and water samples. We also analyzed the virulent plasmid sequence and identified 2 regions that exhibit a clear geographical variation: a 4243-bp Tn3-like transposon and a 9-bp small sequence repeat (SSR). The Tn3-like transposon was only found in the isolates from Latin Americas (Mexico and 2 Central American countries), but not in SE Asian isolates, we then developed PCR methods to characterize AHPND-V. parahaemolyticus isolates as either Mexican-type or SE Asian-type based on the presence of the Tn3-like transposon. The SSR is found within the coding region of a hypothetical protein and has either 4, 5, or 6 repeat units. SSRs with 4 repeat units were found in isolates from Vietnam, China, and Thailand. SSRs with 5 repeat units were found in some Vietnamese isolates, and SSRs with 6 repeat units were only found in the Mexican isolates.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Han JE, Tang KFJ, Tran LH, Lightner DV (2015) Photorhabdus insect related (Pir) toxins-like genes in a plasmid of Vibrio parahaemolyticus, the causative agent of acute hepatopancreatic necrosis disease (AHPND) of shrimp. Dis Aquat Org. 113: 33-40.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Han JE. Tang KFJ, Pantoja CR, White BL, Lightner DV (2015) qPCR assay for detecting and quantifying a virulence plasmid in acute hepatopancreatic necrosis disease (AHPND) due to pathogenic Vibrio parahaemolyticus. Aquaculture 442: 12-15.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Naim S, Tang KFJ, Yang M, Lightner DV, Nibert ML (2015) Extended genome sequences of penaeid shrimp infectious myonecrosis virus strains from Brazil and Indonesia. Archive of Virology 160: 1579-1583.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Han JE, Tang KFJ, Pantoja CR, Mohney L, Lightner DV (2015) Plasmid mediated tetracycline resistance of Vibrio parahaemolyticus associated with acute hepatopancreatic necrosis disease (AHPND). Aquaculture Reports 2: 17-21.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Tang KFJ, Pantoja CR, Redman RM, Han JE, Tran LH, Lightner DV (2015) Development of in situ hybridization and PCR assays for the detection of Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp. J Invertebrate Pathology 130: 37-41.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Lee C-T, Chen I-T, Yang Y-T, et al. (2015) The opportunistic marine pathogen Vibrio parahaemolyticus becomes virulent by acquiring a plasmid that expresses a deadly toxin. PNAS 112: 10798-10803
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Han JE, Tang KFJ, Lightner DV (2015) Genotying of the virulence plasmid harbored in the Vibrio parahaemolyticus isolates causing acute hepatopancreastic necrosis disease (AHPND) in shrimp. Dis Aquat Org. 115: 245-251.
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Progress 10/01/13 to 09/30/14
Outputs
Target Audience: The global (Latin American, SE Asia, South Asian, East African, and Middle Eastern) shrimp farming industries. Changes/Problems: Most of our work was completed in 2013. Hence, in 2014 work had to progress to include topics like EMS/AHPND. What opportunities for training and professional development has the project provided? None. How have the results been disseminated to communities of interest? Yes and the publications listed have been distributed to interested countries including China, Vietnam, Thailand, Malaysia, Mexico and the Kingdom of Saudi Arabia where EMS/AHPND could become a major disease. What do you plan to do during the next reporting period to accomplish the goals? We will continue to do research and development work on EMS/AHPNS/D in order to: 1. Identify the toxin gene (is it a bacteriophage or a plasmid?) present in the strains of AHPND that cause the severe sloughing of HP tubule epithelial cells in the acute phase of the disease. 2. Better define laboratory challenge methods with Vibrio parahaemolyticus (VP) so that more uniform results can be obtained from studies in which juvenile shrimp (mostly Penaeus vannamei) are given a challenge dose of VP. 3. Perform a bacterial growth study for VP in which optical densities (taken early, mid and late in the exponential growth phase, during the stationary phase, and during the death phase of a culture of the agent) are compared graphically to total aerobic plate counts. 4. Refine the PCR method for detection of the disease to include a qPCR method.
Impacts
What was accomplished under these goals?
Early Mortality Syndrome or Acute Hepatopancreatic Necrosis Disease was first recognized in Vietnam, but it subsequently spread to Mexico, Guatemala, and Nicaragua where it typically causes nearly 100% losses in affected shrimp farms. EMS/AHPND will become an OIE listed disease in May of 2015. It is because of EMS/AHPND for which my lab's research focus has shifted to the disease caused by certain specific strains of Vibrio parahaemolyticus.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Nunan, L., Lightner, D., Pantoja, C., and Gomez-Jimenez, S. 2014. Detection of acute hepatopancreatic necrosis disease (AHPND) in Mexico. Dis Aquat Org 111:81-86.
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Tang, FJ. Tang and D.V. Lightner. 2014. Homologues of insecticidal toxin complex genes within a genomic island in the marine bacterium Vibrio parahaemolyticus. FEMS Microbiol Lett (2014) pp 1-9.
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Tsai, Y.-L., Wang, H.-C., Lo, C-F., Tang-Nelson, K., Lightner, D., Ou, B.-R., Tsai, C.-F., Grace Cheng, H.-F., Teng, P.-H., Lee, P.-Y. 2014. Validation of a commercial insulated isothermal PCR-based POCKIT test for rapid and easy detection of White Spot Syndrome Virus infection in Litopenaeus vannamei. Plos One, vol. 9, pp.1-8.
- Type:
Other
Status:
Published
Year Published:
2014
Citation:
Tran, L.H., Fitzsimmons, K.M. and Lightner, D.V. 2014. Tilapia could enhance water conditions, help control EMS in shrimp ponds. January/February 2014; Global Aquaculture Advocate.
- Type:
Other
Status:
Published
Year Published:
2014
Citation:
Tran, L.H., Fitzsimmons, K. and Lightner, D.V. 2014. March/April 2014, AQUA Culture Asia Pacific Magazine.
|
Progress 01/01/13 to 09/30/13
Outputs
Target Audience: The global (Latin American, SE Asia, South Asian, East African, and Middle Eastern) shrimp farming industries. Changes/Problems: Most of the objectives (e.g. objectives 1-4) have been completed, and we have published papers on each of the principal topics. Hence, when EMS/AHPND emerged in SE Asia, my lab was very well positioned to devote much of our effort in this newly emerging disease. What opportunities for training and professional development has the project provided? Last year 1 PhD student from Ho Chi Minh City in Vietnam did his doctoral dissertation on EMS/AHPNS/D. In 2014, I have hired a postdoctoral research associate to continue work on EMS/AHPHS/D. How have the results been disseminated to communities of interest? Yes. Four popular press articles and one journal paper have been published. These are available to the international shrimp farming community. What do you plan to do during the next reporting period to accomplish the goals? We will continue to do research and development work on EMS/AHPNS/D in order to: 1. Identify the toxin gene (is it a bacteriophage or a plasmid?) present in the strains of AHPND that cause the severe sloughing of HP tubule epithelial cells in the acute phase of the disease. 2. Better define laboratory challenge methods with Vibrio parahaemolyticus (VP) so that more uniform results can be obtained from studies in which juvenile shrimp (mostly Penaeus vannamei) are given a challenge dose of VP. 3. Perform a bacterial growth study for VP in which optical densities (taken early, mid and late in the expotential growth phase, during the stationary phase, and during the death phase of a culture of the agent) are compared graphically to total aerobic plate counts. 4. Refine the PCR method for detection of the disease to include a qPCR method.
Impacts
What was accomplished under these goals?
Most of my lab's work in 2013 was on a newly emerged disease in SE Asian shrimp farms (China, Malaysia, Vietnam and Thailand) and its recent occurrence in western Mexico (Mexican States of Nayarit, Sinaloa, and Sonora). The disease is called Early Mortality Syndrome, but it is more correctly called Acute Hepatopancreatic Necrosis Syndrome/Disease (AHPND) because of the severe sloughing of hepatopancreatic (HP) tubule epithelial cells from the HP tubules in the acute phase. In the terminal phase of the disease, the HP is destroyed by invasive Vibrio spp. which utilize the sloughed and necrotic HP tubule cells as a nutrient source. Since its appearance in 2009, the disease has cost the SE Asian (and Mexico in 2013) in excess of USD$1 billion each year. Because of shortages of farmed shrimp, prices are expected to nearly double in 2014. This lab developed a case definition for the disease which was presented to the Network of Aquaculture Centers in Asia (NACA) at a conference in Bangkok, Thailand in 2012. That case definition led to the discovery of the causative agent by this laboratory in March of 2013. An abstract from a recent key publication on EMS/AHPND is as follows: A new emerging disease in shrimp, first reported in 2009, was initially named Early Mortality Syndrome (EMS). In 2011, a more descriptive name for the acute phase of the disease was proposed as Acute Hepatopancreatic Necrosis Syndrome (AHPNS). Affecting both Pacific white shrimp (Penaeus vannamei) and black tiger shrimp (Penaeus monodon), the disease has caused significant losses in Southeast Asian shrimp farms. AHPNS was first classified as idiopathic because no specific causative agent had been identified. However, since early in 2013, the Aquaculture Pathology Laboratory at the University of Arizona (UAZ-APL) was able to isolate the causative agent of AHPNS in pure culture. Immersion challenge tests were employed for infectivity studies, which induced 100% mortality with typical AHPNS pathology to experimental shrimp exposed to the pathogenic agent. Subsequent histological analyses showed that AHPNS lesions were experimentally induced in the laboratory and were identical to those found in AHPNS infected shrimp samples collected from the endemic areas. Bacterial isolation from the experimentally infected shrimp enabled recovery of the same bacteria colony type found in field samples. In three separate immersion tests, using the recovered isolate from the AHPNS positive shrimp, the same AHPNS pathology was reproduced in experimental shrimp with consistent results. Hence, AHPNS has a bacterial etiology and Koch’s Postulates have been satisfied in laboratory challenge studies with the isolate of the agent. The agent is a unique strain of Vibrio parahaemolyticus.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Aranguren L.F., M. Salazar, K. Tang, X. Caraballo and D. Lightner. 2013. Characterization of a new strain of Taura syndrome virus (TSV) from Colombian shrimp farms and the implication in the selection of TSV resistant lines. J. of Invertebrate Pathology 112: 68-73.
Nunan, L.M., C.R. Pantoja, S.Gomez-Jimenez and D.V. Lightner. 2013. Candidatus Hepatobacter penaei, an intracellular pathogenic bacterium enteric in the hepatopancreas of the marine shrimp Penaeus vannamei (Crustacea:Decopoda). Applied and Environmental Microbiology 79(4): 1407-1409.
Tang, K.F.J., C.R. Pantoja, R.M. Redman and D.V. Lightner. 2013. A histological variant of white spot syndrome virus (WSSV) from the Kingdom of Saudi Arabia. J. of Invertebrate Pathology 113: 82-85.
Tran, Loc, L. Nunan, R.M. Redman, L.L. Mohney, C.R. Pantoja, K. Fitzsimmons and D.V. Lightner. 2013. Determination of the infectious nature of the agent of acute hepatopancreatic necrosis syndrome affecting penaeid shrimp. Diseases of Aquatic Organisms 105: 45-55.
Tang, K.F.J., M. Le Groumellec and D.V. Lightner. 2013. Novel, closely related, white spot syndrome virus (WSSV) genotypes from Madagascar, Mozambique and the Kingdom of Saudi Arabia. Diseases of Aquatic Organisms 106: 1-6.
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: Objective 1. A new strain of Taura syndrome virus (TSV) was characterized during the reporting period. This new strain is from the Kingdom of Saudi Arabia (KSA) and the strain is called SA/10 or SA/11, depending on the year the isolates were obtained. This new isolate came from significant disease outbreaks in the Indian white shrimp, Penaeus (Fenneropenaeus) indicus, being grown in intensive culture systems in KSA. Sequencing the capsid protein was completed, and the SA/10 and SA/11 isolates constitute a new genotype of TSV. A paper describing this new TSV genotype was published (Tang et al. 2012). When considered with the other known genotypes of TSV, the KSA strains brings the TSV genotype total to six. Objective 2. Work on this objective was completed prior to the 2011 reporting period and we published a paper describing a PCR method to distinguish infectious IHHNV from the integrated form of the virus, which apparently has been integrated into the genome of Penaeus monodon from the western Indian Ocean for possibly millions of years. Objective 3. UAZ has developed an IMN disease challenge model in which a known amount of IMNV is injected into the abdomen of the test shrimp. While this methods seems to give different survival values when numerous families of P. vannamei are challenged, it does bypass potential innate immunity that may be a factor in shrimp aquaculture where per os infection via cannibalism is the most likely route of transmission of IMNV. UAZ completed its work to determine if a per os IMN challenge method can be developed, and we have concluded that the injection challenge method for determining family line resistance to IMNV is superior to the per os challenge methods so far tested. Objective 4. Work on Penaeus vannamei nodavirus (PvNV) was completed during the reporting period and our findings were reported by Tang et al. (2011). Objective 5. A new disease emerged in SE Asia in both P. monodon and P. vannamei farms during the reporting period. The disease is called "Early Mortality Syndrome." The disease has two distinct phases. The first is called acute hepatopancreatic necrosis syndrome and it is characterized by significant sloughing of hepatopancreatic epithelial cells. The second and typically terminal phase of the disease is a massive bacterial (most isolates obtained are Vibrio harveyi or V. parahaemolyticus). While UAZ has carried out numerous studies on this emerging disease, its etiology remains unknown. The cost to the shrimp farming industries of China, Malaysia, Vietnam and Thailand now exceeds $1 billion and the losses to the industry continue to mount. PARTICIPANTS: Training or professional development opportunities: The UAZ Aquaculture Pathology Laboratory (UAZ-APL) has offered for the past 24 years an annual Summer Session Course called the Shrimp Pathology Short Course. We have had over 700 participants from 59 countries attend our Short Course. The 2013 Shrimp Pathology Short Course is scheduled for June 3-14, 2013. In addition to our on-campus 2-week Shrimp Pathology Short Course, the UAZ-APL also offers special shrimp pathology short courses and workshops at international sites. These special courses and workshops run from 1 day to 10 days in duration, and we have offered these courses in 17 countries and we have had well over 1500 participants. TARGET AUDIENCES: Our target audience is the national and the global shrimp shrimp farming industries. PROJECT MODIFICATIONS: In 2013 the UAZ Aquaculture Pathology Lab (UAZ-APL) will continue to investigate new strains of TSV as they become apparent globally. This will be accomplished by sequencing capsid protein 2 (CP2) and comparing the resultant sequence to the other six known genotypes of TSV. We will also change much of our research focus to the emerging shrimp disease in SE Asia which is adversely affecting exports of farmed shrimp from the affected countries (China, Malaysia, Vietnam and Thailand) to markets in the USA. The disease is called EMS, for early mortality syndrome. Shrimp can suffer very high mortalities due to EMS within the first 30 days after stocking of a typical shrimp pond. We anticipate that other diseases of farmed shrimp are likely to emerge in 2013, which UAZ-APL will investigate.
Impacts
Objective 1. Investigate the genetic diversity of different isolates of TSV as a function of origin and epidemiology of these isolates. The emergence of a new strain of TSV brings to six the number of known strains of this highly pathogenic virus disease. The success of selective breeding for TSV resistance by the now defunct US Marine Shrimp Farming Consortium (in which UAZ was a member and responsible for much of the selection of TSV resistant stocks) has resulted in selected family lines of Penaeus vannamei that exhibit very good resistance to all of the known strains of TSV. Objective 2. To determine flanking sequence the IHHNV-related sequence, and to develop a specific PCR assay to distinguish this apparently non-infectious variety from infectious strains of the virus. UAZ developed a method to distinguish the integrated form of IHHNV in the genome of Penaeus monodon from the infectious strains of the virus. This has been a welcome development in the domestic and international industries that are developing SPF domesticated lines of P. monodon because lines with the integrated non-infectious form of IHHNV can be distinguished and are not killed because they give a positive PCR test for IHHNV. Objective 3. To develop a laboratory challenge model for IMNV in which selected shrimp stocks are tested for resistance to known amounts of the virus. The development of a per os IMNV challenge model, to supplement our current injection model, is in progress. Injection assays in which shrimp from various families are challenged with an infected inoculum with a high count of IMNV were found to be superior to per os assays because with infection of the virus we can obtain a prevalence of infection approaching 100% in a 20 day assay, while per os infection success is often less than 50% in a 50 day assay. Objective 4. To determine the complete genomic sequence of PvNV (Penaeus vannamei nodavirus), analyze the organization of the viral genome, characterize the phylogenetic relationship of this virus with other members of the family Nodaviridae, and determine if the PvNV can infect Macrobrachium rosenbergii. This objective has been completed. While PvNV is of some interest to the shrimp farming industry in the Americas, the disease caused by PvNV is no where near as serious or significant as the OIE listed disease (white tail disease) caused by MrNV. Objective 5. To determine the nature of new or emerging bacterial or fungal diseases in domestically farmed shrimp and to develop case definitions for these diseases so that methods for their diagnosis and detection of their etiological agent(s) can be developed. EMS is an emerging disease that is causing severe losses to the shrimp farming industries of China, Vietnam, Malaysia and in Thailand beginning in early 2012. The disease has the potential to negatively impact shrimp breeding companies in Hawaii and Florida that export broodstock to these affected countries,and possibly more significantly to affect the prices paid by importers of SE Asian farmed shrimp. With external funding from FAO and companies in SE Asia, UAZ is working to determine the etiology of the disease.
Publications
- Nunan, L.M., C.R. Pantoja, S.Gomez-Jimenez and D.V. Lightner. 2013. Candidatus Hepatobacter penaei, an intracellular pathogenic bacterium enteric in the hepatopancreas of the marine shrimp Penaeus vannamei (Crustacea:Decopoda). Applied and Environmental Microbiology 79(4): 1407-1409.
- Stentiford, G.D., D.M. Neil, E.J. Peeler, J.D. Shields, H.J. Small, T.W. Flegel, J.M. Vlak, B. Jones, F. Morado, S. Moss, J. Lotz, L. Bartholomay, D.C. Behringer, C. Hauton, and D.V. Lightner. 2012. Disease will limit future food supply from the global crustacean fishery and aquaculture sectors. Journal of Invertebrate Pathology 110: 141-157.
- Tang, K.F.J., S.A. Navarro, C.R. Pantoja, F.L. Aranguren and D.V. Lightner. 2012. New genotypes of white spot syndrome virus (WSSV) and Taura syndrome virus (TSV) from the Kingdom of Saudi Arabia. Diseases of Aquatic Organisms 99: 179-185.
- Tang, K.F.J., C. R. Pantoja and D.V. Lightner. 2012. New WSSV, TSV Genotypes Identified In Saudi Arabia. Global Aquaculture Advocate May/June, pp. 20-21.
- Wyban, J., B. Noble and D.V. Lightner. 2012. Performance testing on SPF shrimp lines. Aquaculture Asia Pacific, July/August, 8(4): 18-20.
- Aranguren L.F., M. Salazar, K. Tang, X. Caraballo and D. Lightner. 2013. Characterization of a highly pathogenic strain of Taura syndrome virus (TSV) from Colombian shrimp farms. J. of Invertebrate Pathology 112:68-73.
- Cuellar-Anjel, J., B.White-Noble, P.Schofield, R. Chamorro and D.V. Lightner. 2012. Report of significant WSSV resistance in the Pacific White shrimp, Litopenaeus vannamei, from a Panamanian breeding program. Aquaculture 368-369: 36-39.
- Lightner, D.V., R.M. Redman, C.R. Pantoja, K.F.J. Tang, B.L. Noble, P. Schofield, L.L. Mohney, L.M. Nunan and S.A. Navarro. 2012. Historic emergence, impact and current status of shrimp pathogens in the Americas. Journal of Invertebrate Pathology 110: 174-183.
- Lightner, D.V. 2012. Global transboundry disease politics: the OIE perspective. Journal of Invertebrate Pathology 110: 184-187.
- Lightner, D.V., R.M. Redman, C.R. Pantoja, B.L. Noble and L. Tran. 2012. Early mortality syndrome affects shrimp in Asia. Global Aquaculture Advocate January/February, pp. 40.
- Moss, A.M., D.R. Moss, S.M. Arce, D.V. Lightner, and J.M. Lotz. 2012. The role of selective breeding and Biosecurity in prevention of disease in penaeid shrimp aquaculture. Journal of Invertebrate Pathology 110: 247-250.
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: Objective 1. A new strain of TSV was characterized during the reporting period. This new strain is called "TSV CO10" because it was derived from Colombian stocks of farmed Pacific white shrimp, Penaeus vannamei. The sequencing of the entire ssRNA genome was completed, but the main difference was found in capsid protein 2 (CP2). The sequence of CP2 from TSV CO10 was found to differ from other TSV isolates from the Americas by 4%. In experimental infection studies, the pathogenicity of TSV CO10 was also found to differ from other strains of TSV from the Americas and to cause mortalities in challenged unselected P. vannamei at a rate comparable to the Belize strain of TSV (TSV BZ 02). A paper reporting this new strain of TSV is in preparation. Objective 2. Work on this objective was completed prior to the reporting period. A PCR method was developed and internally validated which distinguishes infectious IHHNV from the IHHNV-related sequence that is integrated into the genome of Penaeus monodon from the western Indian Ocean. In addition to the method being published after discovery of the integrated sequence in P. monodon (Tang et al. 2007), the method has been shown to be valid by labs in Taiwan and Thailand and it will be published in the 2012, 7th Edition of OIE Manual of Diagnostic Tests for Aquatic Animals (OIE 2012) as the recommended method for distinguishing infectious IHHNV from this non-infectious integrated form. Objective 3. UAZ has developed an IMN disease challenge model in which a known amount of IMNV is injected into the abdomen of the test shrimp. While this methods seems to give different survival values when numerous families of P. vannamei are challenged, it does bypass potential innate immunity that may be a factor in shrimp aquaculture where per os infection via cannibalism is the most likely route of transmission of IMNV. UAZ is working with a domestic breeding company in Hawaii to determine if a per os IMN challenge method can be developed. Objective 4. Additional sequence information on PvNV was developed and reported (Tang et al. 2011). The PvNV genome consists of two segments: RNA1 (3111 bp) and RNA2 (1183bp). RNA1 contains 2 overlapped open reading frames (ORF A and B) which may encode for an RNA-dependent polymerase (RdRp) and a B2 protein. RNA2 contains a single ORF that may encode the viral capsid protein. Objective 5. A new disease emerged in SE Asia in both P. monodon and P. vannamei farms during the reporting period. The disease is called "Early Mortality Syndrome," but its etiology remains unknown. In some preliminary work funded by the OIE (World Animal Health Organization) and Uni-President Feeds of Vietnam, we determined that EMS is not infectious and that its agent appears to be a toxicant. With additional funding anticipated from CP Foods of Thailand, China and Vietnam, we plan to work with a Dr. Shane Snyder's laboratory on campus do isolate the putative toxin(s) and confirm that it (they) are the cause of EMS using bioassays with live shrimp. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Objective 1. Investigate the genetic diversity of different isolates of TSV as a function of origin and epidemiology of these isolates. The emergence of a new strain of TSV brings to six the number of known strains of this highly pathogenic virus disease. The success of selective breeding for TSV resistance by the now defunct US Marine Shrimp Farming Consortium (in which UAZ was a member and responsible for much of the selection of TSV resistant stocks) has resulted in selected family lines of Penaeus vannamei that exhibit very good resistance to all of the known strains of TSV. Objective 2. To determine the 3' end flanking sequence the IHHNV-related sequence (type B, which is integrated into the genome of P. monodon), and to develop a specific PCR assay to distinguish this apparently non-infectious variety from infectious strains of the virus. UAZ developed a method to distinguish the integrated form of IHHNV in the genome of Penaeus monodon from the infectious strains of the virus. This has been a welcome development in the domestic and international industries that are developing SPF domesticated lines of P. monodon because lines with the integrated non-infectious form of IHHNV can be distinguished and are not killed because they give a positive PCR test for IHHNV. Objective 3. To develop a laboratory challenge model for IMNV in which selected shrimp stocks are tested for resistance to known amounts of the virus. The development of a per os IMNV challenge model, to supplement our current injection model, is in progress. Objective 4. To determine the complete genomic sequence of PvNV (Penaeus vannamei nodavirus), analyze the organization of the viral genome, characterize the phylogenetic relationship of this virus with other members of the family Nodaviridae, and determine if the PvNV can infect Macrobrachium rosenbergii. This objective has been completed. While PvNV is of some interest to the shrimp farming industry in the Americas, the disease caused by PvNV is no where near as serious or significant as the OIE listed disease (white tail disease) caused by MrNV. Objective 5. To determine the nature of new or emerging bacterial or fungal diseases in domestically farmed shrimp and to develop case definitions for these diseases so that methods for their diagnosis and detection of their etiological agent(s) can be developed. EMS is an emerging disease that is causing severe losses to the shrimp farming industries of China, Vietnam and Malaysia. The disease has the potential to negatively impact shrimp breeding companies in Hawaii and Florida that export broodstock to these affected countries. With external funding, UAZ is working to determine the etiology of the disease.
Publications
- Lightner D.V. 2011. Virus diseases of farmed shrimp in the Western Hemisphere (the Americas): a review. Journal of Invertebrate Pathology 106: 110-130.
- Tang, K.F.J., C.R. Pantoja, R.M. Redman, S.A. Navarro and D.V. Lightner. 2011. Ultrastructural and sequence characterization of Penaeus vannamei nodavirus (PvNV) from Belize. Diseases of Aquatic Organisms 94: 179-187.
- Stentiford, G.D. and D.V. Lightner. 2011. Cases of White Spot Disease (WSD) in European shrimp farms. Aquaculture 319: 302-306.
- Lightner D.V. and R.M. Redman. 2010. The global status of significant infectious diseases of farmed shrimp. Asian Fisheries Society Journal 23: 383-426. Aranguren L.F., K.F.J. Tang and D.V. Lightner. 2010. Quantification of the bacterial agent of necrotizing hepatopancreatitis (NHP-B) by real-time PCR and comparison of survival and NHP load of two shrimp populations. Aquaculture 307: 187-192.
- Tang, K.F.J., G.A. Messick, C.R. Pantoja, R.M. Redman, and D.V. Lightner. 2011. Histopathological characterization and in situ detection of Callinectes sapidus reovirus. Journal of Invertebrate Pathology 108: 226-228.
- Lightner, D.V. 2011. Status of shrimp diseases and advances in shrimp health management. pp. 121-134, in M.G. Bondad-Reantaso, J.B. Jones, F. Corsin and T. Aoki (editors), Diseases in Asian Aquaculture VII. Fish Health Section, Asian Fisheries Society, Selangor, Malaysia. 385 p.
- Nunan, L.M. and D.V. Lightner. 2011. Optimized PCR assay for detection of white spot syndrome virus (WSSV). Journal of Virological Methods 171: 318-321.
- Heres, A., R. Redman and D.V. Lightner. 2011. Histopathology of Spiroplasma penaei systemic infection in experimentally infected Pacific white shrimp, Penaeus vannamei. The Israeli Journal of Aquaculture Bamidgeh IIC:63: 589-596.
- Bakonyi, T., B.C. Bonning, D.V. Lightner, Y.J. Chen, N. Nakashima, S.M. Valles and P. Christian. 2012. Dicistroviridae. In A.M.Q. King, M.J. Adams, E.B. Carstens and E.J. Lefkowitz, editors: Virus Taxonomy. 9th Report of the International Committee on Taxonomy of Viruses, Oxford, Elsevier, pp. 840 - 845.
- Aoki, T., J.R. Bonami, D.V. Lightner, C.F. Lo, G.D. Stentiford, J.H. Leu, J. M. Vlak, K. Soderholl, P.J. Walker, T.W. Flegel, H.C. Wang, X. Xu, and H. Yang. 2012. Nimaviridae. In A.M.Q. King, M.J. Adams, E.B. Carstens and E J. Lefkowitz, editors: Virus Taxonomy. 9th Report of the International Committee on Taxonomy of Viruses, Oxford, Elsevier, pp. 229 - 234.
- Schneemann, A., J.R. Bonami, D.V. Lightner, K.L. Johnson, T. Nakai and R. Thiery. 2012. Nodaviridae. In A. M.Q. King, M.J. Adams, E.B. Carstens and E.J. Lefkowitz, editors: Virus Taxonomy. 9th Report of the International Committee on Taxonomy of Viruses, Oxford, Elsevier, pp. 1061 - 1068.
- Bakonyi, T., B.C. Bonning, D.V. Lightner, Y.J. Chen, N. Nakashima, S.M. Valles and P. Christian. 2012. Iflaviridae. In Andrew M.Q. King, Michael J. Adams, Eric B. Carstens, and Elliot J. Lefkowitz, editors: Virus Taxonomy. 9th Report of the International Committee on Taxonomy of Viruses. Oxford. Elsevier , pp. 846 - 849.
- Bonami, J.R., D.V. Lightner, E.J. Snijder, J.A. Cowley, P.J. Walker, R. J. de Groot and T.W. Flegel. 2012. Roniviridae. In A.M.Q. King, M.J. Adams, E.B. Carstens and E.J. Lefkowitz, editors: Virus Taxonomy. 9th Report of the International Committee on Taxonomy of Viruses. Oxford, Elsevier, pp. 829 - 834.
- Lightner D.V. 2012. Diagnostic Manual for Aquatic Animal Diseases. 7th Edition. World Organization for Animal Health ( Office International des Epizooties = OIE), Paris: 1)Disinfection of Crustacean Farms, Section C of Methods for Disinfection of Aquaculture Establishments; 2)Diseases of Crustaceans, General Information; 3) Infectious Hypodermal and Hematopoietic Necrosis; 4)Infectious Myonecrosis; 5)Taura Syndrome; 6)Tetrahedral Baculoviruosis (Baculovirus penaei); and 7)Spherical Baculovirosis (Penaeus monodon-type baculovirus).
- Lightner D.V. 2011. Aquatic Animal Health Code. 14th Edition. Office International des Epizooties (OIE), Paris:1)Infectious hypodermal and haematopoietic necrosis, Chapter 9.2.; 2)Infectious myonecrosis, Chapter 9.3.; 3)Necrotizing Hepatopancreatitis, Chapter 9.4.; 4)Taura syndrome, Chapter 9.5.; 5)White spot disease, Chapter 9.6.; 5)White tail disease, Chapter 9.7.; 6)Yellow head disease, Chapter 9.8. (Available on-line at http://www.oie.int/international-standard-setting/aquatic-code/access -online/.
- Aranguren L.F., M. Salazar, K. Tang and D. Lightner. 2012. Characterization of a highly pathogenic strain of Taura syndrome virus (TSV)from Colombian shrimp farms. J. Virus Research.
- Lightner, D.V. and R.M. Redman. 2012. Development of specific pathogen-free (SPF) shrimp stocks and their application to sustainable shrimp farming. Woodhead Publishing, LTD., Cambridge. UK.
- Aranguren L.F., K. Tang and D. Lightner. 2012. Protection from yellow head virus (YHV) infection in Penaeus vannamei pre-infected with Taura syndrome virus (TSV). Diseases of Aquatic Organisms.
- Lightner, D.V. 2010. Crustacean (penaeid shrimp) diseases. Annual Reports of OIE Reference Laboratories and Collaborating Centres for 2010. Office International des Epizooties, Paris.
- Tang K.F.J., and D.V. Lightner. 2012. Duplex real-time PCR for detection and quantification of monodon baculovirus (MBV) and hepatopancreatic parvovirus (HPV) in Penaeus monodon. Diseases of Aquatic Organisms.
- Lightner, D.V. 2011. Enfermedades virales en camarones cultivados en el hemisferio oeste: una resena. Panorama Acuicola 16(4): 8-20.
- Nunan, L.M. and D.V. Lightner. 2011. One-step PCR for white spot syndrome detection. Global Aquaculture Advocate. January/February 2011, pp. 74.
- Arce, S.M., S.M. Moss and D.V. Lightner. 2011. Biosecurity principles for sustainable production using SPF shrimp. Global Aquaculture Advocate. May/June 2011, pp. 14-16.
- Pantoja, C.R., S.A. Navarro and D.V. Lightner. 2011. Ring test compare PCR results from diagnostic laboratories. Global Aquaculture Advocate. May/June 2011, pp. 36-37.
- Cuellar-Anjel, J., R. Chamorro, B.White-Noble, P.Schofield and D.V. Lightner. 2011. Testing finds resistance to WSSV in shrimp from Panamanian breeding program. Global Aquaculture Advocate July/August 2011, pp. 65-66.
- Cuellar-Anjel, J., R. Chamorro, B.White-Noble, P.Schofield and D.V. Lightner. 2011. White shrimp Litopenaeus vannamei significantly resistant against WSSV. Panorama Acuicola 16(5): 46-48.
- Cuellar-Anjel, J., B.White-Noble, P.Schofield R. Chamorro and D.V. Lightner. 2011. Selected families of white shrimp Litopenaeus vannamei from Panama breeding program show important resistance to WSSV. November/December AQUA Culture Asia Pacific Magazine, pp. 36-37.
- Lightner, D.V. and R.M. Redman. In press. Global health issues - crustaceans. Proceedings of the Global Conference on Aquatic Animal Health Programmes, Their Benefits for Global Food Security. Panama City, Panama on 28-30 June 2011. OIE, Paris.
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