scholarly journals In-vitro studies on antiviral effects of Galaxaura elongata marine algae on white spot syndrome virus

2018 ◽  
Vol 34 (1) ◽  
pp. 162-171 ◽  
Author(s):  
Ahmed Abdel-Wahab
Keyword(s):  
Marine Algae ◽  
White Spot Syndrome Virus ◽  
In Vitro Studies ◽  
White Spot ◽  
Antiviral Effects ◽  
White Spot Syndrome

scholarly journals Identification of a Shrimp E3 Ubiquitin Ligase TRIM50-Like Involved in Restricting White Spot Syndrome Virus Proliferation by Its Mediated Autophagy and Ubiquitination

2021 ◽  
Vol 12 ◽  
Author(s):  
Chao Zhao ◽  
Chao Peng ◽  
Pengfei Wang ◽  
Lulu Yan ◽  
Sigang Fan ◽  
...  
Keyword(s):  
White Spot Syndrome Virus ◽  
Penaeus Monodon ◽  
White Spot ◽  
Positive Role ◽  
Protein Levels ◽  
Mrna And Protein Expression ◽  
White Spot Syndrome ◽  
Critical Components ◽  
Trim Proteins

Most tripartite motif (TRIM) family proteins are critical components of the autophagy machinery and play important roles in host defense against viral pathogens in mammals. However, the roles of TRIM proteins in autophagy and viral infection have not been studied in lower invertebrates, especially crustaceans. In this study, we first identified a TRIM50-like gene from Penaeus monodon (designated PmTRIM50-like), which, after a white spot syndrome virus (WSSV) challenge, was significantly upregulated at the mRNA and protein levels in the intestine and hemocytes. Knockdown of PmTRIM50-like led to an increase in the WSSV quantity in shrimp, while its overexpression led to a decrease compared with the controls. Autophagy can be induced by WSSV or rapamycin challenge and has been shown to play a positive role in restricting WSSV replication in P. monodon. The mRNA and protein expression levels of PmTRIM50-like significantly increased with the enhancement of rapamycin-induced autophagy. The autophagy activity induced by WSSV or rapamycin challenge could be inhibited by silencing PmTRIM50-like in shrimp. Further studies showed that rapamycin failed to induce autophagy or inhibit WSSV replication after knockdown of PmTRIM50-like. Moreover, pull-down and in vitro ubiquitination assays demonstrated that PmTRIM50-like could interact with WSSV envelope proteins and target them for ubiquitination in vitro. Collectively, this study demonstrated that PmTRIM50-like is required for autophagy and is involved in restricting the proliferation of WSSV through its ubiquitination. This is the first study to report the role of a TRIM family protein in virus infection and host autophagy in crustaceans.

scholarly journals White Spot Syndrome Virus Open Reading Frame 222 Encodes a Viral E3 Ligase and Mediates Degradation of a Host Tumor Suppressor via Ubiquitination

2006 ◽  
Vol 80 (8) ◽  
pp. 3884-3892 ◽  
Author(s):  
Fang He ◽  
Beau J. Fenner ◽  
Andrew K. Godwin ◽  
Jimmy Kwang
Keyword(s):  
Tumor Suppressor ◽  
Cell Line ◽  
White Spot Syndrome Virus ◽  
Ovarian Cancer Cell Line ◽  
E3 Ligase ◽  
Cancer Cell Line ◽  
White Spot ◽  
White Spot Syndrome

ABSTRACT We have characterized a white spot syndrome virus (WSSV) RING-H2-type protein, WSSV222, which is involved in ubiquitination. WSSV222 exhibits RING-H2-dependent E3 ligase activity in vitro in the presence of the specific conjugating enzyme UbcH6. Mutations in the RING-H2 domain abolished WSSV222-dependent ubiquitination, revealing the importance of this domain in WSSV222 function. Yeast two-hybrid and pull-down analyses revealed that WSSV222 interacts with a shrimp tumor suppressor-like protein (TSL) sharing 60% identity with human OVCA1. To better characterize the interaction of WSSV222 and TSL in vivo, we established a stable TSL-expressing cell line derived from the human ovarian cancer cell line A2780, where we observed a TSL-dependent prolonged G1 phase. Furthermore, we detected WSSV222-mediated ubiquitination and MG132-sensitive degradation of TSL both in shrimp primary cell culture and in the TSL-expressing cell line. Transient expression of TSL in BHK cells leads to apoptosis, which was rescued by WSSV222. Taken together, our data suggest that WSSV222 acts as an antiapoptosis protein by ubiquitin-mediated proteolysis of TSL to ensure successful WSSV replication in shrimp.

scholarly journals A gC1qR Prevents White Spot Syndrome Virus Replication in the Freshwater Crayfish Pacifastacus leniusculus

2010 ◽  
Vol 84 (20) ◽  
pp. 10844-10851 ◽  
Author(s):  
Apiruck Watthanasurorot ◽  
Pikul Jiravanichpaisal ◽  
Irene Söderhäll ◽  
Kenneth Söderhäll
Keyword(s):  
Cell Culture ◽  
Viral Replication ◽  
White Spot Syndrome Virus ◽  
Pacifastacus Leniusculus ◽  
White Spot ◽  
Tissue Cell ◽  
Freshwater Crayfish ◽  
White Spot Syndrome

ABSTRACT The gC1qR/p32 protein is a multiple receptor for several proteins and pathogens. We cloned a gC1qR homologue in a crustacean, Pacifastacus leniusculus, and analyzed the expression of P. leniusculus C1qR (PlgC1qR) in various tissues. The gC1qR/p32 transcript was significantly enhanced by white spot syndrome virus (WSSV) infection 6 h after viral infection both in vitro in a hematopoietic tissue cell culture (Hpt) and in vivo compared to appropriate controls. Moreover, PlgC1qR silencing in both the Hpt cell culture and live crayfish enhanced the WSSV replication. In addition, by making a recombinant PlgC1qR protein we could show that if this recombinant protein was injected in a crayfish, Pacifastacus leniusculus, followed by injection of WSSV, this significantly reduced viral replication in vivo. Furthermore, if the recombinant PlgC1qR was incubated with Hpt cells and then WSSV was added, this also reduced viral replication. These experiments clearly demonstrate that recombinant PlgC1qR reduce WSSV replication both in vivo and in vitro. The results from a far-Western overlay and glutathione S-transferase pull-down assays showed that PlgC1qR could bind to VP15, VP26, and VP28. Altogether, these results demonstrate a role for PlgC1qR in antiviral activity against WSSV.

scholarly journals PENGKLONAN GEN PENYANDI VIRAL PROTEIN 15 (VP-15) WSSV DAN APLIKASINYA SEBAGAI VAKSIN REKOMBINAN PADA UDANG WINDU

2018 ◽  
Vol 13 (1) ◽  
pp. 57
Author(s):  
Andi Parenrengi ◽  
Sri Redjeki Hesti Mulyaningrum ◽  
Andi Tenriulo ◽  
Agus Nawang
Keyword(s):  
Nucleotide Sequence ◽  
Viral Protein ◽  
White Spot Syndrome Virus ◽  
Penaeus Monodon ◽  
Recombinant Vaccine ◽  
White Spot ◽  
Tiger Shrimp ◽  
White Spot Syndrome ◽  
Insert Region

Infeksi white spot syndrome virus (WSSV) dapat menyebabkan kematian massal pada budidaya udang windu Penaeus monodon di Indonesia. Infeksi yang terjadi secara sistematis tersebut disebabkan oleh peran gen nucleocapsid viral protein (VP-15). Upaya pengembangan gen VP-15 WSSV untuk menginduksi respons imun dan menetralisasi terhadap infeksi WSSV pada udang windu perlu dilakukan. Penelitian ini bertujuan untuk mengisolasi dan merekombinasikan gen penyandi VP-15 WSSV sebagai vaksin dsRNA, serta menganalisis aplikasinya pada udang windu. Gen VP-15 diisolasi dari udang windu yang terinfeksi WSSV, dikloning ke dalam suatu vektor dan ditransformasikan ke sel kompeten (bakteri Escheria coli DH5a). Plasmid diisolasi untuk mengonfirmasi insert region gen VP-15 melalui sekuensing nukleotida. Pembuatan vaksin rekombinan dilakukan secara in-vitro menggunakan kit MEGAscript RNAi dan diaplikasikan ke udang windu melalui metode injeksi dengan dosis tunggal 0,2 µg dan kontrol (tanpa injeksi vaksin). Hewan uji yang digunakan berukuran panjang 14,75±3,17 g dan bobot 11,64±0,76 cm; serta dipelihara pada wadah bak fiber volume 250 L dengan kepadatan 10 ekor/bak. Hasil penelitian menunjukkan bahwa gen penyandi VP-15 telah diisolasi dari udang windu dan vaksin rekombinan telah dihasilkan secara in-vitro. Analisis sekuens nukleotida memperlihatkan bahwa sisipan gen DNA VP-15 sebesar 253 bp dan menunjukkan kemiripan yang tinggi (99%) pada GenBank. Penggunaan vaksin rekombinan dsRNA dengan dosis 0,2 µg memperlihatkan sintasan udang windu yang dapat mencapai 40,0% dibandingkan dengan kontrol hanya 3,3% (peningkatan 36,7%). Gambaran histopatologi pada jaringan hepatopankreas udang windu pada perlakuan kontrol menunjukkan adanya kerusakan inti sel, akibat infeksi WSSV. Gene VP-15 berpotensi sebagai bahan vaksin rekombinan dsRNA dalam mencegah infeksi WSSV.Infection of white spot syndrome virus (WSSV) causes bulk mortalities of tiger shrimp Penaeus monodon cultured in Indonesia. The nucleocapsid viral protein-15 (VP-15) is strongly suspected to be responsible for the systemic infection of WSSV. The development of VP-15 WSSV gene for inducing the immune response to and neutralize WSSV infection of tiger shrimp is vitally needed. The aim of this study was to isolate and clone the gene encoding VP-15 WSSV as dsRNA vaccine and assess the vaccine application to tiger shrimp. VP-15 gene was isolated from the genomic DNA of infected tiger shrimps, cloned into the vector, and transformed into competent cells (Escheria coli DH5a). The plasmid was isolated to confirm the insert region gene of VP-15 by the nucleotide sequence. Production of dsRNA vaccine was performed by in-vitro using MEGAscript RNAi kit and applied to tiger shrimp through muscular injection at a single dosage of 0.2 µg and without dsRNA as a control treatment. The average size of tiger shrimps used was 14.75±3.17 g in weight and 11.64±0.76 cm in length and stocked in 250 L fiber tank at 10 ind./tank. The results of the study showed the VP-15 gene was successfully isolated from the tiger shrimps and the recombinant vaccine was produced by in-vitro. The analysis of nucleotide sequence showed that the inserted DNA was 253 bp and showed a high similarity (99%) with VP-15 gene deposited in the GenBank. The application of dsRNA vaccine showed that the dosage of 0.2 ¼g resulted in the survival rate of 40.0% compared with without dsRNA (control) of 3.3% (36.7% increment). Hepatopancreas histology indicated obvious damages to cell nucleus in the un-vaccinated tiger shrimp caused by the virus infection. We suggest that the VP-15 gene is a very promising dsRNA recombinant vaccine against WSSV infection.

scholarly journals Viral ubiquitin ligase WSSV222 is required for efficient white spot syndrome virus replication in shrimp

2009 ◽  
Vol 90 (6) ◽  
pp. 1483-1490 ◽  
Author(s):  
Fang He ◽  
Syed Musthaq Syed ◽  
A. S. Sahul Hameed ◽  
Jimmy Kwang
Keyword(s):  
Virus Replication ◽  
White Spot Syndrome Virus ◽  
Tumour Suppressor ◽  
White Spot ◽  
Time To Death ◽  
Real Time Quantitative Pcr ◽  
Efficient Replication ◽  
White Spot Syndrome ◽  
Apoptosis Regulation

The E3 ligase WSSV222 of white spot syndrome virus (WSSV) is involved in anti-apoptosis regulation by ubiquitin-mediated degradation of tumour suppressor-like protein (TSL), a shrimp tumour suppressor. In the present study, WSSV222 gene expression was silenced by using specific small interfering RNA (siRNA) in Sf9 and BHK cells. Based on the results of the in vitro silencing, WSSV-challenged shrimp were treated with anti-WSSV222 siRNA to knock down WSSV222 protein expression. The survival rate of shrimp and the efficiency of WSSV replication were assessed to evaluate the efficacy of anti-WSSV222 siRNA in regulating WSSV infection in shrimp. The anti-WSSV222 siRNA reduced the cumulative mortality in shrimp challenged with 103 copies of WSSV and delayed the mean time to death in shrimp challenged with the higher dose of 106 copies. The results of real-time quantitative PCR showed that virus replication was delayed and reduced in WSSV-challenged shrimp treated with anti-WSSV222 siRNA in comparison with challenged shrimp treated with random-control siRNA. Co-immunoprecipitation assays revealed that WSSV222 silencing inhibited the degradation of TSL in WSSV-challenged shrimp, indicating the requirement for WSSV222 for efficient replication of WSSV in shrimp.

In vitro white spot syndrome virus (WSSV) replication in explants of the heart of freshwater crab, Paratelphusa hydrodomous

2012 ◽  
Vol 183 (2) ◽  
pp. 186-195 ◽  
Author(s):  
K.S. Nathiga Nambi ◽  
S. Abdul Majeed ◽  
N. Sundar Raj ◽  
G. Taju ◽  
N. Madan ◽  
...  
Keyword(s):  
White Spot Syndrome Virus ◽  
White Spot ◽  
Freshwater Crab ◽  
White Spot Syndrome

A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus

2007 ◽  
Vol 146 (1-2) ◽  
pp. 96-103 ◽  
Author(s):  
Li Yuan ◽  
Xiaohua Zhang ◽  
Mingxian Chang ◽  
Chensong Jia ◽  
Sean M. Hemmingsen ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
White Spot Syndrome Virus ◽  
Neutralization Assay ◽  
White Spot ◽  
White Spot Syndrome

scholarly journals Phycoerythrin from Colaconema sp. Has Immunostimulatory Effects on the Whiteleg Shrimp Litopenaeus vannamei and Increases Resistance to Vibrio parahaemolyticus and White Spot Syndrome Virus

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2371
Author(s):  
Po-Tsang Lee ◽  
Jing Huang ◽  
Chin-Yi Huang ◽  
Zi-Xuan Liu ◽  
Han-Yang Yeh ◽  
...  
Keyword(s):  
Litopenaeus Vannamei ◽  
Vibrio Parahaemolyticus ◽  
White Spot Syndrome Virus ◽  
Expression Profiles ◽  
White Spot ◽  
Shrimp Culture ◽  
White Spot Syndrome ◽  
Whiteleg Shrimp

We investigated whether phycoerythrin (PE), a pigment sourced from marine algae, could act as an immunomodulatory agent in whiteleg shrimp (Litopenaeus vannamei). To this end, PE was extracted and purified from a PE-rich macroalgae, Colaconema sp. Our in vitro analysis demonstrated that PE enhanced prophenoloxidase and phagocytosis activity but inhibited the production of reactive oxygen species in hemocytes. Additionally, the PE signal could be detected using an in vivo imaging system after its injection into the ventral sinus of the cephalothorax of whiteleg shrimp. The expression profiles of fourteen immune-related genes were monitored in hemocytes from whiteleg shrimp injected with 0.30 μg of PE per gram of body weight, and crustin, lysozyme, penaiedin 4, and anti-lipopolysaccharide factor showed up-regulated post-stimulation. The induction of immune genes and enhancement of innate immune parameters by PE may explain the higher survival rates for shrimp that received different doses of PE prior to being challenged with Vibrio parahaemolyticus or white spot syndrome virus compared to controls. Combined, these results show that PE from Colaconema sp. can differentially stimulate the immune response of whiteleg shrimp in vitro and in vivo and could potentially be used as an immunomodulator in shrimp culture.

scholarly journals Antilipopolysaccharide Factor Interferes with White Spot Syndrome Virus Replication In Vitro and In Vivo in the Crayfish Pacifastacus leniusculus

2006 ◽  
Vol 80 (21) ◽  
pp. 10365-10371 ◽  
Author(s):  
Haipeng Liu ◽  
Pikul Jiravanichpaisal ◽  
Irene Söderhäll ◽  
Lage Cerenius ◽  
Kenneth Söderhäll
Keyword(s):  
Cell Culture ◽  
White Spot Syndrome Virus ◽  
Pacifastacus Leniusculus ◽  
White Spot ◽  
Freshwater Crayfish ◽  
Hematopoietic Tissue ◽  
Viral Loads ◽  
White Spot Syndrome

ABSTRACT In a study of genes expressed differentially in the freshwater crayfish Pacifastacus leniusculus infected experimentally with the white spot syndrome virus (WSSV), one protein, known as antilipopolysaccharide factor (ALF), was chosen, among those whose transcript levels increased upon viral infection, for further studies. ALF RNA interference (RNAi) experiments in whole animals and in cell cultures indicated that ALF can protect against WSSV infection, since knockdown of ALF by RNAi specifically resulted in higher rates of viral propagation. In a cell culture of hematopoietic tissue (Hpt) from P. leniusculus, quantitative PCR showed that knockdown of ALF by RNAi resulted into WSSV levels that were about 10-fold higher than those treated with control double-stranded RNA (dsRNA). In addition, RNAi experiments with other crayfish genes that had been found to be up-regulated by a WSSV infection did not result in any changes of viral loads. Thus, the cell culture does not respond to dsRNA in a similar manner, as shown earlier for dsRNA injected into shrimp, which gave a higher degree of resistance to WSSV infection. If ALF transcription in whole animals was stimulated by the administration of UV-treated WSSV, a partial protection against a subsequent challenge with the active virus was conferred to the host. This is the first crustacean gene product identified with the capacity to interfere with replication of this important pathogen.

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