Progress 01/01/24 to 12/31/24
Outputs
Target Audience:White Spot Disease (WSD), caused by the White Spot Syndrome Virus (WSSV) is a threat to crustacean farming (e.g., shrimp & crawfish) in the US and elsewhere in the world causing billions of dollars of losses. Currently, there isno commercially available therapeutics against the disease. The goal of this project was todevelopan oral vaccine against WSD in shrimp and other crustaceans. The research findings of this project will benefit shrimp and other crustacean farming industry andaquafeed industry producing formulated diet for shrimp farmingin the US and elsewhere in the world. Additionally, the research finding will be immensely beneficial toprofessions engaged in infectious disease research and extension activities involving crustaceans, shellfish and fish inacademia, government and non-governmental agencies worldwide. Changes/Problems:Challenges faced with this project: Technical Cloning of some hairpin RNA in the shrimp nodaviral vector were not successful. For example, as described under the Objective #1, cloning of hairpin RNA targeting the WSSV genes, VP19 and a dual hairpin RNA VP28+ VP09 could not be achieved. This is not uncommon since clones of some RNA with extensive secondary structures are not stable in the vector they are cloned. Lack of a WSSV LD50 model posed a technical challenge in determining the efficacy of RNAi-based therapeutics in shrimp. There are published papers describing LD50 model for WSSV via injection route of virus challenge. However, injection does not simulate the natural routes of infection (i.e., oral and immersion roues) andthere is no robust LD50 model for WSSV experimental challenge using an oral route of WSSV infection. Researchers routinely used oral challenges using WSSV-infected tissue at 2.5 to5% of the biomass of the tank to attain WSSV infection in helathy animals. Our initial experimental challenge (i.e., an oral challenge using 2.5% of the biomass)in screening shrimp nodaviral vector containing candidate RNAi constructs resulted in 100% mortality of animals in all treatments (See accomplishment under Objective #3). Subsequently, we used WSSV-infected tissue at 1.5% of the biomass of the tank. Although this resulted a 100% cumulative mortality in most WSSV-challenged tanks, there was a delay in mortality in some RNAi treatment (e.g., VP09 construct), and there was a significant difference in WSSV quantitative load in the animals that scummed to death in some treatments compared to others (i.e., higher WSSV load in shrimp in GFP control treatments andVP28 RNAicompared to VP28 injected animals, VP28-U6 construct and VP09 constructs). This shows an optimized LD50 model for WSSV infection using an oral route of virus challenge is needed for screening RNAi-based as well as otehr therapeutics. Non-technical (Personnel change):Apostdoctoral fellow and the Co-PI of the project, Dr. Rod R. Russel Allenton left and took a job in a biotech company in Tokyo, Japan after about 1.5 years into the project. This resulted in a delay in accomplishing the goals. Subsequently, Dr. Hung N. Mai, a Research Scientist in Dhar's lab, and another postdoctoral fellow, Dr. Dinh-Hung Nguyen joined the project and completed the tasks. What opportunities for training and professional development has the project provided?The project provided opportunities for training oftwo Postdoctoral Fellows, Dr.Rod Russel R. Alenton and Dr. Nguyen Dinh-Hung to workon shrimp virology, molecular biology and theraputic developmes in crusatceans. Additionally, a research scientist, Dr. Hung N. Mai working in the Aquaculture Pathology Laboratory was also involved in the project. The PI of the project, Dr. Arun K. Dhar, the Postdoctoral Fellows and the Research Scientists had the opportunity to attend national/ international conferneces and meetings to share their findings and gain insights of the participants through in-person and virtual interactions. The project also provided unique opportunities to undergraduate students working in Dr. Dhar's lab to learn cell culture and molecular biology skill while working alongside thePostdoctoral Fellows and Scientists. How have the results been disseminated to communities of interest?The research efforts of the project have been communicated through peer reviewed publication, presentations in international scientific conferneces, invited talks in universities worldwide. We have reached students and researchers internationally through invited talks at universities in Asia (i.e., India and Taiwan),the Americas (i.e., USA and Mexico), . Most target audiences of theproject were reached by presenting at strategic scientific conferences, such as Aquaculture America (2022,2023, 2024)and Conference for Research in Animal Diseases (2023), which bring together universities, industry, government,and non-governmental organizations working on shrimp and VMOs, aquatic animal health professionals. In addition,during the the Aquaculture Pathology laboratory- University of Arizona, Annual Shrimp pathology Short Course, we reached out to attendedd from around the world as well as representatives of government agencies and private companies from different countries worldwide. What do you plan to do during the next reporting period to accomplish the goals?The project ended in December 31, 2024, and this is the final report of the project.
Impacts
What was accomplished under these goals?
Objective #1 : Construction of a shrimp viral vector containing hairpin-RNA (hRNA) targeted against WSSV structural and nonstructural genes. We developed aMacrobrachium rosenbergiinodavirus (MrNV)-based viral vector using a baculovirus expression systemand insect cell culture. The viral vector enables us tocloneand express heterologous RNA, and delivery of the payload (i.e. functional mRNA, RNAi sequence)in crustacean cells via injection and oral routes (i.e. mixing the shrimp viral vector cell culture material with commercial diet).We demonstrated that the vector was capable of delivering a marker gene, GFP in shrimp via injection and per os. The viral vectorwas found to be replication incompetent and did not cause any mortality or pathological manifestation in shrimp when delivered viainjection or oral route. These findings were published in the journalProc. Natl. Acad. Sci- Nexus, 2: 1-9,https://doi.org/10.1093/pnasnexus/pgad278. This was the first a major development in the project and this opened up an avenue for us to try to deliver therapeutic RNA using this viral vector in shrimp via an oral route thorugh commercial diet. Then using this shrimp viral vector, we made four different constructs: a construct carrying GFP gene to monitor the delivery of the payload by the vector; a hairpin RNA targeting a white spot syndrome virus (WSSV) structural gene and a major antigen, VP28 along with GFP,a different hairpin loop of VP28, and a hairpin loop targetingVP9 gene, a non-structural gene of WSSV. We chose these target genes of WSSV (i.e., VP28 and VP9) based on published literature where researchers showed that injection of hairpin RNA/ dsRNAtargeting these genes provides protection against WSSV in shrimp. We made the following constructs using MrNV-based shrimp viral vector: MrNV-GFP, MrNV GFP+VP28, MrNV-VP28, MrNV-VP9. However, our effort to clone adualhairpin RNAs in a single construct,VP28 + VP9 DNA was not successful, likely due to the instability of the construct in the viral vector. Also, our effort to clone a hairpin RNA targeting a different non-structural gene of WSSV, VP19 was also not successful suggesting a need for a further optimization of hairpin RNA cloning in the shrimp viral vector. Overall, the data obtained in accomplishing Objective #1 gave us confidence that cloning of hairpin RNA targeting WSSV gene in shrimp viral vector is feasible, although not all hairpin RNA are stable. Nevertheless, with the constructs we made, we went on to further characterize them and evaluating their efficacy in WSSV control via oral delivery (See accomplishments of Objectives #2 and #3). Objective #2 : Confirm expression of hRNA molecules delivered using a shrimp viral vector in vitro (using Sf9 cells and shrimp primary hemocytes) and in vivo (live shrimp). We accomplished this objective using the recombinant shrimp viral vectorcarrying long hairpin (lh) RNA targeting VP28 from Sf9 cells that are expressing the shrimp viral vector. Transmission electron microscopy (TEM) confirmedthe assembly of viralvector and hairpin cargo for lhVP28 construct.We analyzed thefull-length expression of therapeutic RNA from the viral vector construct,VP28 hRNA using conventional PCR. The expression of MrNV capsid protein gene (which is a part of the shrimp MrNV viral vector) and VP28 RNA were detected by RTqPCR in Sf9 cells confirming the production of the therapeutic RNAmolecule. Objective #3 : Determining the efficacy of shrimp viral vector in delivering hRNA via diet and protecting shrimp against WSD. a. Feed preparation and palatability test:We used commercial shrimp feed to mix with to viral vector solution (1:0.5 ratio of feed weight to viral vector solution in mL) to determine palatability of the feed to shrimp. Food pellets were fully consumed by shrimp within 2-5 mins indicating that the shrimp viral vector containing therapeutic made in insect cell culture solution could be added to commercial shrimp diet without any negative effect on palatability. b. Bioassay #1: Determining the efficacy of shrimp viral vector VP28-hRNA in protecting Specific Pathogen Free (SPF) Penaeus vannamei shrimp against WSSV: Delivery of RNAi via injection followed by an WSSV oral challenge: Survival assay was performed by injecting therapeutic RNA followed by an oral WSSV challenge at 2.5% of the tank biomass. Treatments included: (a) the purified viral vector carrying lhVP28, (b) purified RNA from the produced viral vector, (c) 2% saline (positive control), (d) GFP RNA-treated (negative control), and (e) naïve group. While group a & b provided 100% protection for 14 days post-WSSV challenge, the control treatments (c & d) didn't provide any protection against WSSV. WSSV load was quantified by real-time PCR targeting VP644 gene (unrelated to knockdown gene VP28) and the relative quantification of RNA of the target gene. WSSV could not be detected in a &b compared to the 2x10^5 average viral copy number per nanogram DNA in control treatments. The VP28 RNA level was significantly downregulated in treatments a and b compared to GFP RNA treated group. The data showed RNAi delivered using a shrimp viral vector via injection could protect shrimp from WSSV infection. With these findings, we moved on to delivering RNAi through oral route by mixing with commercial diet followed by an oral challenge. c. Bioassay #2: Delivery of RNAi via oral route followed by a WSSV oral challenge: We used four different constructs to conduct this experiment: MrNV-GFP, MrNV-GFP+VP28, MrNV-VP28, MrNV-VP9. We grew four recombinant viruses in Sf9 cell culture, and the cell culture biomass were mixed with commercial shrimp diet. The experimental diets were fed to SPFPenaeus vannameishrimp for 14 days and then the animals were challenged with WSSV at 2.5% of the biomass in the tank. There were 100% mortality in all four treatments following an oral delivery of the therapeutic RNAi molecules followed by an oral WSSV challenge. We concluded the viral challenge was so lethal that it overwhelmed the host and RNAi delivered via an oral route failed to protect the animals. Bioassay #3:We did a bioassay using WSSV infected tissue at1.5% of the biomass of the tank. We kept an injection treatment group for MrNV-GFP+VP28 construct. The cumulative mortality by 8 days post-challenge reached 100% in all four orally delivered therapeutic group. However, there was a delay in mortality in MrNV-VP9 treatment. Additionally, when MrNV-GFP+VP28 construct was injected 24-hr prior to challenge with WSSV, there was a 100% protection. The result was the same as observed in the Experimental bioassay #1. We found WSSV load was lowest in MrNV-GFP+VP28 injection treatment. WSSV load was also significantly lower in MrNV-VP9 treatment compared to MrNV-GFP and MrNV-VP28 treatments. But there was no significant difference in WSSV load between MrNV-VP9 and MrNV-GFP+VP28. In the MrNV-GFP+VP28 construct, RNAi sequence is under the control of an U6 promoter, compared to polyhedrin (pol) promoter in MrNV-VP28. The U6 promoter is a eukaryotic promoter and likely to be more functional in shrimp compared to pol.We concluded that when the RNAi level in shrimp is high, it will protect the animal from WSSV infection. Future work should focus on optimizing the level of RNAi needed to protect shrimp from WSSV and also optimize promoter in the shrimp viral vector to attain a higher level of RNAi expression. To summarize, we developed a shrimp viral vector that can deliver heterologous RNA (i.e., functional RNA, RNAi sequence) in shrimp. RNAi delivered via injection and using the shrimp viral vector can provide complete protection against WSSV. However, an oral delivery of the RNAi molecules using the shrimp viral vector need further optimization to attain protection against WSSV. The shrimp viral vector is generic in nature and could potentially be used to deliver RNAi molecules to protect shrimp against other viruses.
Publications
- Type:
Other
Status:
Published
Year Published:
2025
Citation:
Dhar, Arun K. 2025. Disease diagnosis and engineering a viral vector for an oral delivery of therapeutic RNA in shrimp , ICAR-Central Institute of Fisheries Education, Kolkata, West Bengal, India, February 25, 2025.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2025
Citation:
Dhar, Arun K. 2025. Leveraging Technology for Genetic Improvement in Aquaculture Production in the Technical Session of the theme Blue Revolution: Harnessing Science and Technology . XVII Agricultural Science Congress, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, India, February 20- 22, 2025.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2025
Citation:
Dhar, Arun K. 2025. Comprehensive monitoring of animal health and developing oral therapeutics are needed for sustainability of shrimp farming worldwide , 14th Asian Fisheries and Aquaculture Forum (14AFAF), Greening the Blue Growth in Asia Pacific February 12-15, 2025, New Delhi, India.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2025
Citation:
Dhar, Arun K. 2025. Engineering a viral vector for the delivery of therapeutic RNA in invertebrates . ICAR-National Institute for Plant Biotechnology, February 09, 2025, New Delhi, India.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Dhar, Arun K. and Thales P. De Andrade. 2024. Current and Emerging Diseases in shrimp: Their Diagnosis and Prevention. XX International Shrimp Farming Symposium, FENECAM 2024, November 19-22, Natal, Brazil.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Alenton, R. R.R., Mai, H. N. & Dhar, A. K. 2024. A replication-deficient shrimp viral vector engineered for the delivery of therapeutic RNA. The Conference of Research Workers in Animal Diseases, January 20-23, 2024, Chicago, Illinois.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Alenton, R. R.R., Mai, H. N. & Dhar, A. K. 2023. An oral RNA delivery platform using reverse-engineered Nodavirus for marine shrimp Aquaculture America 2023 Conference, World Aquaculture Society, February 23-26, 2023, New Orleans, Louisiana.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Alenton, R. R.R., Mai, H. N. & Dhar, A. K. 2023. 2023. Overcoming roadblocks for RNA-targeted antiviral therapy in shrimp through a vector-based oral delivery platform. The Conference of Research Workers in Animal Diseases, January 22-24, 2023, Chicago, Illinois.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Rod Russel R. Alenton., Hung N. Mai, and Arun K. Dhar. 2022. Reverse-genetics approach in developing a
Macrobrachium rosenbergii Nodavirus (MrNV)-based viral vector for an oral delivery of therapeutics in shrimp. Aquaculture
America 2022 Triennial Conference. February 28 - March 4, 2022, Town and Country Conference Center San Diego, CA.
- Type:
Other
Status:
Published
Year Published:
2022
Citation:
Rod Russel R. Alenton. Antiviral therapies for shrimp aquaculture . Ensenada Center for Scientific Research and Higher Education (CISESE), Ensenada, Mexico. August 08, 2022
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Arun K. Dhar. 2022. Emergent Diseases and efforts toward developing antiviral therapy in shrimp . Indian Council of
Agricultural Research Lecture Series on the occasion of 75th Years of Independence of India, organized by Central
Institute of Brackishwater Aquaculture, Chennai, India, July 29, 2022.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2022
Citation:
Rod Russel R. Alenton, Mary Beth B. Maningas. Diagnostics and RNAi based therapeutics for the shrimp industry . National Cheng Kung University, Taipei, Taiwan, July 14, 2022.
- Type:
Other
Status:
Other
Year Published:
2022
Citation:
Arun K. Dhar. 2022. Discovering Pathogens and Developing Antiviral Therapies in Marine Shrimp Using a Reverse Genetic Approach , West Bengal University of Fishery & Animal Sciences, Kolkata, West Bengal, India, July 14, 2022.
- Type:
Peer Reviewed Journal Articles
Status:
Published
Year Published:
2023
Citation:
Alenton, Rod Russel R., Mai, Hung N. and Dhar, Arun K. 2023. Engineering a replication incompetent viral vector for delivery of therapeutic RNA in crustacean. Proc. Natl. Acad. Sci- Nexus, 2: 1-9, https://doi.org/10.1093/pnasnexus/pgad278
|
Progress 01/01/23 to 12/31/23
Outputs
Target Audience: Shrimp and crayfish industry in the US and elsewhere in the world. Aquafeed industry working to develop functional feed to control white spot disease in crustaceans. Animal health professionals working in government, academia, and industry working to develop therapeutics against viral diseases in crustaceans. Changes/Problems:We encountered a technical challenge in cloning a hairpin RNA in the shrimp viral vector. Initially, to expedite the project, the cloning ofhairpin RNA in a shrimp viral vectorwas outsourced to a vendor who routinely provide services of viral vector cloning. Unfortunately, the vendor failed to make stable clone containing therapeutic RNA molecule targeting white spot syndrome virus. Then apostdoctoral fellow working on the project had to clone the gene in hpuse through iterations and improvizing cloning technique. This delayed the project outcomes. So far, have made one out of three viral clones originally proposed in the project proposal. This clone was tested in an injection bioassay to determine the efficacy in providing protection against WSD. We got succesful result in the injection bioassay and we are now cloning the remaining two therapeutic RNA in the shrimp viral vector. What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?The research findings of this project was communicated through a peer-reviewed publication in the journal Proceedings of National Academy of Science-Nexus and through oral presentations in several national and international conferences/ meetings worldwide. The audience of these conferences and meetings included professionals from academic institutions, industry, government, and non-governmental organizations working on shrimp and VMOs, aquatic animal health professionals. A complete list of publications is provided under "Publication" Section. In addition, the findings of this project was also disseminated during a "Shrimp Pathology Short Course", a summer course held in the Aquaculture Pathology Laboratory, School of Animal & Comparative Biomedical Sciences, University of Arizona in June 12-17, 2023. The training was attended by 12 participants from five countries including Brazil, Colombia, Mexico, Venezuela, and USA . What do you plan to do during the next reporting period to accomplish the goals?1. So far we have made recombinant shrimpnodaviral vector carrying hairpin RNA for one of the three targets of WSSV, VP28 gene. We plan to complete the cloning and production ofrecombinant shrimpnodaviral vectorcarrying hairpin RNA for the remaining two target genes of WSSV. 2. An injection bioassay using purified recombinant shrimpnodavirus carrying hairpin RNA targeting WSSV VP28 gene provided control against white spot disease. We will now conduct a bioassay by delivering the recombinant shrimpnodaviruscarrying hairpin RNA targeting WSSV VP28 gene via commercial diet and determine the efficacy in protecting shrimp against WSD. 3. Based on the finding of this bioasay, as decribed in #2, that involves delivering WSSV VP28 RNAi therapeutic via commercial diet, we will design experimental bioasay to deliver the remaining two RNAi constructs via oral route and determine which of three RNAi construct provides highest level of protection against WSD.
Impacts
What was accomplished under these goals?
Objective #1 : Construction of a shrimp viral vector containing hairpin-RNA (hRNA) targeted against WSSV structural and nonstructural genes. Major activities completed / experiments conducted Bivalent hairpin RNA was constructed to investigate the combined strength of dual-targeted hairpin RNAs; VP9 (non-structural) and VP28 + VP9 DNA templates. These need to be clones in BV-MrNV backbone. Also ordered custom synthesis of VP9 (non-structural) and VP28 + VP9 RNA that could be used as positive control for injection purpose. RNA detection, Viral particle isolation and transmission electron microscopy (TEM) confirming the assembly of viral vector and hairpin cargo for lhVP28 construct. Data collected: TEM micrograph showed the successful assembly of the viral vector particles. RNA isolation and visualization displayed a full-length hairpin RNA. Summary statistics and discussion of results Detection of the intact hairpin RNA and TEM micrograph showed the successful production of the viral vector and hairpin cargo complex. Cloning of hairpin RNA targeting VP 19 and Vp9 are on-going. Objective #2 : Confirm expression of hRNA molecules delivered using a shrimp viral vector in vitro (using Sf9 cells and shrimp primary hemocytes) and in vivo (live shrimp). Major activities completed / experiments conducted Analysis of full-length expression of therapeutic RNA from the viral vector construct. (Completed for VP28 hRNA). Data collected: MrNV and VP28 RNA were also detected by RTqPCR in Sf9 cells confirming the production of the therapeutic RNA molecule. RNA extracted from the viral vector and SF9 cells showed the intact hairpin RNA for VP28-hRNA but not for VP28-dsRNA. Objective #3 : Determining the efficacy of shrimp viral vector in delivering hRNA via diet and protecting shrimp against WSD. Major activities completed / experiments conducted Feed preparation and palatability test Trial bioassay run with diet treatments containing viral vector VP28-hRNA and VP28-dsRNA challenged with white spot syndrome virus. Survival assay was performed by direct administration by injecting the purified viral vector carrying lhVP28 and purified RNA from the produced viral vector; followed by a viral challenge test with white spot syndrome virus. Control groups were saline-treated (positive control), GFP RNA-treated (negative control) and naïve groups. Confirmation of the effectiveness of treatment through molecular analysis. Quantification of viral load and knockdown of target viral gene (VP28) after treatment and viral challenge. Data collected: Optimum feed preparation with 1:0.5 ratio (grams to mL) of food pellet to viral vector solution. Food pellets are fully taken up by animals within 2-5 mins. A slight delay (1-2 days) in cumulative mortality was seen in the treatment groups as compared to the control groups. All groups reached 100% mortality. 100% survival for all treatment groups observed for 14 days post-infection. Negative control groups reached 100% mortality at day 6 or 7 post-infection. The viral load was reduced in both treatment groups displaying zero viral DNA copy number. Relative quantification of RNA of the targeted WSSV VP28 gene was confirmed to be downregulated. Summary statistics and discussion of results This mode of preparation retains the integrity and palatability of the food diet ensuring successful oral take-up. A slight delay in mortality may signify that the dosage of the orally delivered therapeutic agent was insufficient, resulting in little to no protection against viral infection. The survival curve shows a significant increase in survival among treatment groups as compared to the negative control groups. Zero copy numbers in treatment groups showed a significant reduction and viral suppression compared to the 2x105 average viral copy number per nanogram DNA in infected shrimp in the negative control. Reduction of viral copies quantifying VP644 gene (unrelated to knockdown gene VP28) displays complete protection from WSSV and not only the knockdown or suppression of one viral gene. Significant downregulation of VP28 RNA in treatment groups as compared to GFP RNA treated groups demonstrates the effectivity of the active component (lhVP28 RNA) of viral vector treatment. Key outcomes or other accomplishments realized. Maintaining the integrity of the food pellet ensures food uptake by the animals. There is a need to concentrate the therapeutic material before incorporating it into the diet and ensure proper dosage rendering protection. The stability of RNA should also be investigated upon incorporation into the feed. Treatment material is capable of suppressing viral infection in shrimp when administered directly by injection. Protection against WSSV resulted from gene-targeted therapy.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Alenton, Rod Russel R., Mai, Hung N. and Dhar, Arun K. 2023. Engineering a replication incompetent viral vector for delivery of therapeutic RNA in crustacean. Proc. Natl. Acad. Sci- Nexus, 2: 1-9, https://doi.org/10.1093/pnasnexus/pgad278
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Aquaculture Pathology Laboratory, University of Arizona - The World Organization for Animal Health Reference Laboratory of Crustacean Diseases Tools, The Launch of a Regional Aquatic Animal Health Laboratory Network (RAAHLN) for Africa, December 05-07, 2023, Pretoria, South Africa.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Dhar, Arun K. 2023. ⿿Keynote address: Expediting pathogen discovery and developing antiviral therapy in shrimp, 4th International mini-Symposium on the Control of Aquatic Animal Disease 2023 (CAAD 2023) will be held at National Cheng Kung University, Taiwan (NCKU) on November 26-29, 2023, Taiwan.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Dhar, Arun K. 2023. ⿿Engineering a Viral Vector Platform for the Delivery of therapeutic RNA in Crustacean & Developing Oral Vaccine in Fish, Global Fisheries Conference, November 21-22, 2023.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Dhar, Arun K. 2023. ⿿Major Diseases & Disease Management in Shrimp Aquaculture, College of Veterinary Medicine, Mississippi State University, Starkville, MS., November 10, 2023.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
8. Dhar, Arun K. 2023. ⿿Diseases & Disease Management in Shrimp Aquaculture, Bandung Institute of Technology (Intitut Teknologi Badung), Jawa Barat, Indonesia, November 01, 2023.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Alenton, R. R.R., Mai, H. N. & Dhar, A. K. 2023. An oral RNA delivery platform using reverse-engineered Nodavirus for marine shrimp Aquaculture America 2023 Conference, World Aquaculture Society, February 23-26, 2023, New Orleans, Louisiana.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Alenton, R. R.R., Mai, H. N. & Dhar, A. K. 2023. 2023. Overcoming roadblocks for RNA-targeted antiviral therapy in shrimp through a vector-based oral delivery platform. The Conference of Research Workers in Animal Diseases, January 22-24, 2023, Chicago, Illinois.
- Type:
Other
Status:
Published
Year Published:
2023
Citation:
1. Dhar, Arun K. 2023. Reverse genetics approaches to study viral pathogenesis and developing viral vector for an oral delivery of therapeutic molecules in shrimp, Central Fisheries Education Institute, as a part of Fulbright Specialist Visit to CIFE, September 09, 2023,
|
Progress 01/01/22 to 12/31/22
Outputs
Target Audience: Shrimp industry in the US and elsewhere in the world. Researchers and professionals from academia, industry, government and non-governmental organizations working on shrimp and fish diseases, and aquatic animal health. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest? The research efforts of the project have been communicated through invited talks in universities worldwide and scientific conferences. We have reached students and researchers internationally through invited talks at universities in Asia (i.e., India and Taiwan) and the Americas (i.e., USA and Mexico). Most target audiences of the project were reached by presenting at strategic scientific conferences, such as Aquaculture America (2022 & 2023) and Conference for Research in Animal Diseases (2023), which bring together universities, industry, government, and non-governmental organizations working on shrimp and VMOs, aquatic animal health professionals. In addition, by holding workshops and training, such as the Aquaculture Pathology Annual Short Course, we reached representatives of government agencies and private companies from different countries in the Americas. What do you plan to do during the next reporting period to accomplish the goals? The next reporting period will cover how our viral vector will be used as a therapeutic oral treatment, which we already started. MrNV viral vector carrying hRNA and dsRNA targeting VP28, VP19, or VP9 shall be incorporated in the diet of shrimp and shall be fed in several dosages; and their successful delivery in the shrimp tissues will be determined and measured. Finally, the efficacy of the treatment materials shall be determined through viral challenge using live animals. In addition to the initial plan, we constructed double-stranded RNA (dsRNA) in addition to hairpin RNA (hRNA). Both RNA forms can trigger gene silencing against viral genes, however, testing both shall give us more options for the stability of the therapeutic materials, which is important for their incorporation into the shrimp diet. During the current reporting period, we experience delays in the synthesizing of the hRNA-carrying viral vector from our designated vendor. For this, we came up with our in-house production which will be easily adapted for additional therapeutic materials that we would need to test.
Impacts
(N/A)
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Rod Russel R. Alenton., Hung N. Mai, and Arun K. Dhar. 2022. Reverse-genetics approach in developing a Macrobrachium rosenbergii Nodavirus (MrNV)-based viral vector for an oral delivery of therapeutics in shrimp. Aquaculture America 2022 Triennial Conference. February 28 - March 4, 2022, Town and Country Conference Center San Diego, CA.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2022
Citation:
Rod Russel R. Alenton Antiviral therapies for shrimp aquaculture . Ensenada Center for Scientific Research and Higher Education (CISESE), Ensenada, Mexico. August 08, 2022
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2022
Citation:
Arun K. Dhar. 2022. Emergent Diseases and efforts toward developing antiviral therapy in shrimp . Indian Council of Agricultural Research Lecture Series on the occasion of 75th Years of Independence of India, organized by Central Institute of Brackishwater Aquaculture, Chennai, India, July 29, 2022.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2022
Citation:
Rod Russel R. Alenton, Mary Beth B. Maningas. Diagnostics and RNAi based therapeutics for the shrimp industry . National Cheng Kung University, Taipei, Taiwan, July 14, 2022.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2022
Citation:
Arun K. Dhar. 2022. Discovering Pathogens and Developing Antiviral Therapies in Marine Shrimp Using a Reverse Genetic Approach , West Bengal University of Fishery & Animal Sciences, Kolkata, West Bengal, India, July 14, 2022.
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