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In Figure a, each column represents the mean of triplicate assay with standard deviation. Asterisks denote significant differences (P<0.01) between samples. All shrimp in the 10 dpi and 15 dpi samples of Bac-wt, Positive control group were died and data is Not Detectable. In Figure b, the arrow marks indicates the expression of VP28 in shrimp tissues.
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White Spot Syndrome Virus (WSSV) is an infectious pathogen of shrimp and other crustaceans, and neither effective vaccines nor adequate treatments are currently available. WSSV is an enveloped dsDNA virus, and one of its major envelope proteins, VP28, plays a pivotal role in WSSV infection. In an attempt to develop a vaccine against WSSV, we insert...
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... In the domain of viral vectors, recombinant baculoviruses have emerged as effective tools for developing viral vector aquaculture vaccines, particularly against emerging viruses like the Cyprinid herpesvirus 2 (CyHV-2) [123], VHSV [124], and the infectious spleen and kidney necrosis virus (ISKNV) [125]. As baculoviruses are known to infect invertebrates, this platform can also be utilized to vaccinate economically important species of aquatic arthropods against severe and detrimental diseases, such as the white spot syndrome (WSS) and nodaviral infections [126][127][128][129]. Nevertheless, baculoviruses can integrate their genomes in the host's chromosomes, thus making their commercial application for vaccine development a near impossible task for aquaculture. ...
... 187 Still, other studies in crustaceans regarding the mechanism of immune action of LGBPs against WSSV infection remain inconclusive, warranting further investigations. [188][189][190] Further, the requirement of Ca 2+ for agglutination efficiency of LGBPs with Gram-positive and Gram-negative bacteria has been reported. 190,191 However, as the binding site is not yet been identified, the mechanism is still under scrutiny. ...
Crustacean species are among the most valued seafood with socioeconomic and ecological implications in the global fisheries sector. Microbial infections to the farming stock due to continued expansion and aquaculture puts global crustacean production at risk. Understanding and reviewing the complex innate immune system of crustaceans would be pivotal in formulating informed decisions for the control and management of epidemic diseases, thus making the global crustacean aquaculture industry sustainable. Under the purview of innate immunity in crustaceans, pattern recognition receptors (PRRs) represent the sensors capable of discriminating non‐self pathogenic modules and invoking the activation of immune responses. The immunity to infections can be manifested at cellular (via phagocytosis, apoptosis, nodulation and encapsulation) and humoral (effected by prophenoloxidase cascade, lectins, antimicrobial proteins) levels leading to the elimination of pathogens or subversion of the pathogenic implications. In this review, we discuss the participatory role of 12 types of PRRs identified from crustacean species with an exhaustive focus on structural and functional specificities in binding to pathogens and possible execution of the immune activity. Moreover, we have elaborated on the PRRs‐associated or possible signalling cascades (Toll‐NFκB, IMD‐NFκB and JAK‐STAT) in crustacean species that could bridge our understanding of the resistance or susceptibility of the host to varied pathogenic infections thus providing clues to the regulatory mechanisms involved in pathogenesis of infectious diseases and mechanisms of immunity.
... Several data types or characteristics have been developed for the prediction of host-pathogen interactions (HPIs), for example, protein sequence similarity [7], gene ontology (GO) annotations [10], protein dominance interactions [8], and protein structural information [11,12]. Interolog and domain-based techniques for the prediction of inter-species PPIs are widely used [6,[13][14][15][16][17]. Many studies have been performed in the past three years to understand the mechanisms of the interactions between SARS-CoV-2 and the human proteome. ...
SARS-CoV-2, a novel betacoronavirus strain, has caused a pandemic that has claimed the lives of nearly 6.7M people worldwide. Vaccines and medicines are being developed around the world to reduce the disease spread, fatality rates, and control the new variants. Understanding the protein-protein interaction mechanism of SARS-CoV-2 in humans, and their comparison with the previous SARS-CoV and MERS strains, is crucial for these efforts. These interactions might be used to assess vaccination effectiveness, diagnose exposure, and produce effective biotherapeutics. Here, we present the HuCoPIA database, which contains approximately 100,000 protein-protein interactions between humans and three strains (SARS-CoV-2, SARS-CoV, and MERS) of betacoronavirus. The interactions in the database are divided into common interactions between all three strains and those unique to each strain. It also contains relevant functional annotation information of human proteins. The HuCoPIA database contains SARS-CoV-2 (41,173), SARS-CoV (31,997), and MERS (26,862) interactions, with functional annotation of human proteins like subcellular localization, tissue-expression, KEGG pathways, and Gene ontology information. We believe HuCoPIA will serve as an invaluable resource to diverse experimental biologists, and will help to advance the research in better understanding the mechanism of betacoronaviruses.
... RNA interference (RNAi) is the process of sequence-specific posttranscriptional gene silencing by which short interfering RNA (siRNA) or short hairpin RNA (shRNA) are used to inactivate the expression of target genes [18,19]. Although several attempts are made to vaccinate crustaceans by administering DNA constructs, which express viral capsid genes [20][21][22], as well as silencing compounds used (namely dsRNA, siRNA and shRNA). The major drawback of this method is that unlike temporal silencing compounds (dsRNA/siRNA/shRNA), DNA transfection might produce a genetically modified organism (GMO). ...
Abstract
In order to understand the TLR pathway defence mechanism of Penaeus monodon, the essential molecule involved in the
transduction of Toll-pathway, Interleukin-1 (IL-1) receptor-associated kinase-4 (IRAK-4) was investigated by the mechanism of RNA
interference (RNAi) by silencing of the mRNA expression of IRAK-4 gene. In this study we have developed short hairpin RNA (shRNA)
construct in pSUPER vector targeting IRAK-4 gene silencing of TLR pathway in P. monodon. The silencing efficiency of pSh-IRAK-4
construct was first confirmed in-vitro in primary haemocyte culture by transfection using pSh-IRAK-4 plasmid, followed by induction
with Lipopolysaccharides (LPS). Loss of expression of IRAK-4 was studied by relative gene expression in pSH-IRAK-4 group compared
to LPS induced group, the maximum suppression of IRAK-4 gene in cell culture was recorded as 96% at 12h and 93.5% at 24h post
LPS induction in pSh-IRAK-4 group. After confirmation of silencing efficiency of construct, the expression of TLR genes of IRAK-4
mediated pathway, was studied post LPS induction both in vitro using real-time qRT-PCR with β-actin as the internal reference gene.
For this IRAK-4 downstream genes TRAF6, Dorsal and 4 Antimicrobial peptides (AMPs) (ALF, PEN, AST, Crustin) molecules were
studied. Significant downregulation of mRNA expression level in downstream molecules of TLR pathway below IRAK-4 gene viz.,
TRAF6, Dorsal, and 4 AMPs (ALF, PEN, AST, Crustin) compared to LPS group in response to LPS stimulation was observed in pShIRAK-4 group (P ≤ 0.05). Taking all these results together, it is confirmed that TLR pathway is governed by central mediator kinase
molecule IRAK-4, when induced by LPS ligand, NF-kB activates the downstream cascades of AMPs of the Toll pathway in P. monodon.
Our result confirms the designing of a novel pSh-IRAK-4 construct and its application in efficient silencing of IRAK-4 gene in P.
monodon. Plasmid-based IRAK-4 knockdown approach would provide an insight to the role of IRAK-4 in shrimp immune system.
Keywords: RNA Interference; pSUPER Vector; shRNA; IRAK-4
... It is reported that crabs can stay longer with latent infection as a carrier of the virus for their high disease resistance capacity and transmit the virus to shrimps [5,6]. 100% of shrimps are reported to die within three to ten days of infection [7]. If shrimps and crabs are cultured together in the improved traditional shrimp ghers, WSSV can be transmitted from crabs to shrimps making shrimps more vulnerable to death due to white spot disease [8]. ...
In this study, previously published Rab7 sequences from National Center for Biotechnology Information (NCBI) have been investigated from chordates, mollusks, annelids, cnidarians, amphibians, priapulids, brachiopods, and arthropods including decapods and other groups. Among decapod crustacean isolates, amino acid variations were found in 13 locations. Penaeid shrimps had variations in positions 13 (I ⟶ J), 22 (T ⟶ A), 124 (G ⟶ X), and 149 (V ⟶ X) while interestingly the freshwater prawn and mitten crab both had amino acid substitutions in positions 87 (V ⟶ C) and 95 (T ⟶ S) along with the other disagreements in amino acid positions 178 (S ⟶ N), 201 (D ⟶ E), 181 (E ⟶ D), 182 (L ⟶ I), 183 (Y ⟶ G), 184 (N ⟶ H), and 198 (A ⟶ T). Among 100 isolates of Rab7 from organisms of various phyla, mutations were observed in several positions. These mutations caused variations in hydrophobicity and isoelectric point which impact the ligand-protein binding affinity. Some common mutations were found in the organisms of the same phylum and among different phyla. Homology modeling of Rab7 proteins from different organisms was done using SWISS-MODEL and validated further by developing Ramachandran plots. Protein-protein docking showed that active residues were there in the binding interfaces of Rab7 from organisms of seven different phyla and VP28 of WSSV. Similarities were observed in the Rab7-VP28 complexes in those selected organisms which differed from the Rab7-VP28 complex in the case of Penaeid shrimp. The findings of this study suggest that WSSV may exist in different marine organisms that have Rab7 protein and transmit to crustaceans like shrimps and crabs which are of commercial importance.
... Intensive aquaculture is mainly the characteristic of single species (Tilman et al., 2006), high-density per unit area, and high yield (Crowder et al., 2010). However, intensive aquaculture reduces the biodiversity of an ecosystem (de Mazancourt et al., 2013) , (Loreau et al., 2001) and results in epidemic diseases and economic losses (Altieri, 2004) (Crowder et al., 2010), especially it is more prominent in invertebrates because of a lack of adaptive immunity (SÁCHEZ-PAZ, 2010); thus, vaccination is not useful in disease control (Musthaq and Kwang, 2011), which leads to large-scale use of chemical substances. In 2011, China's chemical drugs used in aquaculture amounted to 1.3 billion Chinese yuan (American dollars$ 210 million). ...
Seafood security is essential in modern society. In 2013, Bush and colleagues stated, ‘Aquaculture, farming aquatic organisms, provides close to 50% of the world’s supply of seafood, with a value of United States $125 billion. It makes up 13% of the world’s animal-source protein (excluding eggs and dairy) and employs an estimated 24 million people’. With the increase in the human population and reducing fishing resources, humans increasingly rely on aquacultural products as the primary protein sources for many countries. Aquacultural productivity has been improving in recent years, and in certain countries, the aquaculture output is more than the fishing output. For example, Chinese aquaculture production is more than fishing output, which provides one-third of animal protein. Thus, intensive aquaculture has become the main supply with global aquatic products (FAO). In recent years, it is estimated that each person consumption of aquaculture products is 130 kg in some countries (Iceland). Here, we illustrate the road blocker in farmed shrimp production and provide our resolution. The global pandemic of white spot syndrome (WSS), caused by the white spot syndrome virus (WSSV), bears a devastating economic loss in farmed shrimp production, thereby jeopardizing seafood security. Currently, there is no effective control for WSS. Conventional single-species intensive farming removes the spatiotemporal interaction between different species. We hypothesize that establishing the spatiotemporal interface of a predator–prey may control WSS outbreak. We search for the pathways for the mechanisms by which predator–prey species interact and compete across spatial scales to characterize WSSV dispersal at regional scales for the local spatiotemporal structure of viral transmission. Thus, we create a generalizable and turnable engineered ecosystem that provides a clear route to prosperity and well-being to harness the world’s aquatic “blue” food systems to help end hunger.
... The present results raise whether A. mellifera can be protected against natural infections by priming them in their hives. Indeed, there is evidence of successful use of "vaccines" in Crustacea (Kwang, 2011;Valdez et al., 2014;Chang et al., 2018;Johnson et al., 2008;Wang et al., 2019) and Mollusca (Lafont et al., 2017;Yang et al., 2021) against its natural enemies, and maybe, a similar rationale could be used in pollinators (Lanz-Mendoza and Contreras-Garduño, 2021). For example, pollinators could be primed against particles of inactive viruses and, consequently, respond better against lethal doses of the same virus? ...
Invertebrates' immune priming or innate immune memory is an analogous response to the vertebrates' adaptive memory. We investigated if honey bees have immune memory. We compared survival and immune response between bees that were: 1) manipulated (Naïve), 2) challenged twice with the same pathogen Escherichia coli (Memory), 3) challenged twice with different pathogens (Staphylococcus aureus versus E. coli, Micrococcus lysodeikticus versus E. coli), or 4) with PBS (the diluent of bacteria) versus E. coli (heterologous challenge; Control). Results indicate better survival in the Memory than the Control group, and the Memory group showed a similar survival than Naïve insects. The Memory group had higher lytic activity but lower prophenoloxidase, phenoloxidase activity, and hemocyte count than the Control and Naïve groups. No differences were found in relative expression of defensin-1. This first demonstration of immune memory opens the questions about its molecular mechanisms and whether, immune memory could be used against natural parasites that affect honey bees, hence, if they could be “vaccinated” against some natural parasites.
... The critical role of LGBP in the immune system by regulating the activation and/or activity of the prophenoloxidase cascade in P. stylirostris against WSSV has been previously reported [104]. Moreover, in a study to determine the efficacy of a vaccine against WSSV in black tiger shrimp, Syed and Kwang [106] compared the expression level of the LGBP gene in WSSV-treated and Bac-VP28-treated shrimp at different time points. The results showed that LGBP is upregulated in the WSSV-treated shrimp, whereas downregulated in the Bac-VP28-treated shrimp at 7-and 10 days post-infection. ...
Innate immunity is the only defense system for resistance against infections in crustaceans. In crustaceans, white spot diseases caused by white spot syndrome virus (WSSV) are a serious viral disease with high accumulative mortality after infection. Attachment and entry into cells have been known to be two initial and important steps in viral infection. However, systematic information about the mechanisms related to WSSV infection in crustaceans is still limited. Previous studies have reported that cellular receptors are important in the innate immune system and are responsible for the recognition of foreign microorganisms and in the stimulation of the immune responses during infections. In this review, we summarize the current understanding of the functions of cellular receptors, including Toll, C-type lectin, scavenger receptor, β-integrin, polymeric immunoglobulin receptor, laminin receptor, globular C1q receptor, lipopolysaccharide-and β-1,3-glucan-binding protein, chitin-binding protein, Ras-associated binding, and Down syndrome cell adhesion molecule in the innate immune defense of crustaceans, especially shrimp and crabs, in response to WSSV infection. The results of this study provide information on the interaction between viruses and hosts during infections, which is important in the development of preventative strategies and antiviral targets in cultured aquatic animals.
... 59 17 Immersion method is also expensive and a commercial scale implementation is difficult. 62 The oral delivery system has been found to be one of the most feasible methods among others. However, a major disadvantage associated with oral delivery is that all specimens may not receive the antiviral or a proper dosage of it. ...
White spot syndrome virus (WSSV) causes severe disease in crustaceans, specifically shrimp, leading to substantial economic losses to the aquaculture industry. Existing treatment methods fail to sufficiently curb the occurrence and spread of the disease, necessitating the development of effective control and treatment strategies. Structural studies have primarily focused on envelope proteins and a few non‐structural proteins involved in host entry, viral replication, viral assembly and host immune evasion, thus presenting themselves as potential drug targets. Here, we provide a comprehensive review of the insights gained from such structural elucidations to contribute to developing novel therapeutic strategies. In addition, we touch upon the current and future perspectives towards structure‐based drug or vaccine development against WSSV.
... This virus can be transmitted both horizontally and vertically [12,18], and once an outbreak of WSSV occurs, it wipes out entire population in many aquatic farms within a few days [14]. Infection of the WSSV on shrimp is characterized by a rapid mortality up to 100% within 7-10 days [19]. ...
... The circular genome of the WSSV is approximately 275 nm in length and 120 nm in width with tail-like appendages at one end, and composed of five known major structural proteins: VP28, VP19, VP26, VP24 and VP15 [19,21]. Studies on WSSV viral proteins have demonstrated that VP28 and VP19 are associated with the virion envelope [21,22] while VP26 acts as a tegument protein linking the two nucleocapsid-associated proteins VP24 and VP15 to the envelope [17,23]. ...
White Spot Syndrome Virus (WSSV) has emerged as one of the most prevalent and lethal viruses globally, and infects both shrimps and crabs in the aquatic environment. This study aimed to investigate the occurrence of WSSV in different ghers of Bangladesh and the virulence of the circulating phylotypes. We collected 360 shrimp (Penaeus monodon) and 120 crab (Scylla sp.) samples from the South-East (Cox’s Bazar) and South-West (Satkhira) coastal regions of Bangladesh. The VP28 gene-specific PCR assays and sequencing revealed statistically significant (p < 0.05, Kruskal Wallis test) differences in the prevalence of WSSV in shrimps and crabs between the study areas (Cox’s Bazar and Satkhira), and over the study periods (2017-2019). The mean Log load of WSSV varied from 8.40 (Cox’s Bazar) to 10.48 (Satkhira) per gram of tissue. The mean values for salinity, dissolved oxygen, temperature and pH were 14.71±0.76 ppt, 3.7±0.1 ppm, 34.11±0.38˚C and 8.23±0.38, respectively in the WSSV-positive ghers. The VP28 gene-based phylogenetic analysis showed an amino-acid substitution (E→G) at 167th position in the isolates from Cox’s Bazar (referred to as phylotype BD2) compared to the globally circulating one (BD1). Shrimp PL artificially challenged with BD1 and BD2 phylotypes with filtrates of tissue containing 0.423 X 109 copies of WSSV per mL resulted a median LT50 value of 73 hrs and 75 hrs, respectively. The in-vivo trial showed higher mean Log WSSV copies (6.47±2.07 per mg tissue) in BD1 challenged shrimp PL compared to BD2 (4.75±0.35 per mg tissue). Crabs infected with BD1 and BD2 showed 100% mortality within 48 hrs and 62 hrs of challenge, respectively with mean Log WSSV copies of 12.06±0.48 and 9.95±0.37 per gram tissue, respectively. Moreover, shrimp antimicrobial peptides (AMPs) penaeidin and lysozyme expression was lower in BD1 challenged group compared to BD2 challenged shrimps. These results collectively demonstrated that relative virulence properties of WSSV based on mortality rate, viral load and expression of host immune genes in artificially infected shrimp PL could be affected by single aa substitution in VP28.
