Silencing of gonad-inhibiting hormone gene expression in Penaeus monodon by feeding with GIH dsRNA-enriched Artemia

Treerattrakool, Supattra; Chartthai, Chuthakorn; Phromma-in, Nipatthra; Panyim, Sakol; Udomkit, Apinunt

doi:10.1016/j.aquaculture.2013.04.024

Cited by 36 publications

(31 citation statements)

References 17 publications

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“…In contrast, no deaths were reported when dsRNA was orally administered in food with bacteria expressing specific dsRNA (6.0 × 10 10 bacteria/g, extruded feed with β-actindsRNA) (35). Similar results were observed when silencing the viral gene of the associated virus of the gills (GAV) of the giant tiger prawn with bacteria expressing specific dsRNA (4.9 × 10 10 bacteria/g extruded feed of GAV-dsRNA) where survival was 97% by injection and 15-35% by oral administration (36). A similar diminished effect, by applying dsRNA-vp28 (Structural gene of the WSSV virus cell envelope) in the diet was found by Sarathi (7,8) when 25 µg VP28-dsRNA was administered intramuscularly or orally in food supplemented with chitosan and bacteria expressing VP28-dsRNA (1×10 9 bacteria/mL);…”

Section: Discussionsupporting

confidence: 65%

“…Mientras que, no se reportaron mortalidades cuando se administró el dsRNA por vía oral en alimento administrado con bacterias que expresaba dsRNA especifico (6.0 × 10 10 bacterias/g de alimento extruido con β-actin-dsRNA) (35). Resultados similares fueron observados al silenciar el gen viral del virus asociado a las branquias (GAV) del camarón P. monodon con bacterias que expresaban dsRNA especifico (4.9 × 10 10 bacterias/g de alimento extruido con GAV-dsRNA) donde la supervivencia fue del 97% por inyección, y del 15-35% por administración oral (36).…”

Section: Discussionunclassified

“…survival was 65% against challenge with WSSV using injection and 37% survival with oral treatment. Treerattrakool et al (36) silenced the gonad inhibitor hormone (GIH) in the giant tiger prawn when dsRNA-GIH was injected intramuscularly (0.3 μg dsRNA-GIH/g shrimp), but only 50% in shrimp fed a diet containing Artemia salina enriched with the bacteria E. coli dsRNA-GIH (120 OD 600 ). Although the highest antiviral effect observed occurs in organisms to which the specific dsRNA was administered by injection, its usefulness in the field is null because of the high percentage of organisms that are required to be protected, so oral administration remains the best option for dsRNA even though the antiviral effect is lower.…”

Section: Discussionmentioning

confidence: 99%

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Digestibilidad in vitro del dsRNA específico por enzimas del tracto digestivo del camarón Litopenaeus vannamei

2017

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Objective. The digestibility of specific dsRNA by action of the enzymes of digestive tract of the whiteleg shrimp Litopenaeus vannamei was determined in vitro. Materials and methods. Digestive enzyme activity (amylase, lipase, protease, DNase and RNase) was measured in the stomach, digestive gland, and anterior, middle, and posterior intestine of juvenile shrimp, and the digestibility of DNA, RNA and the dsRNA-ORF89, specific to WSSV, was determined by in vitro assays, as well as electrophoretic and densitometric analyses. Results. The highest enzymatic activity was found in the digestive gland: amylase (81.41%), lipase (92.60%), protease (78.20%), DNase (90.85%), and RNase (93.14%). The highest digestive capacity against DNA, RNA, and dsRNA was found in the digestive gland (5.11 ng of DNA per minute, 8.55 ng of RNA per minute, and 1.48 ng dsRNA per minute). Conclusions. The highest digestibility of dsRNA-ORF89, specific to WSSV, was found in the digestive gland, whereas the lowest digestibility was observed in the posterior intestine. This is the first report regarding the digestibility of dsRNA-ORF89 by whiteleg shrimp digestive tract enzymes, with potential therapeutic importance in shrimp culture to prevent WSSV disease through balanced feed.Keywords: Aquaculture, balanced feed, digestive gland, nucleic acids, WSSV (Sources: MeSH, FAO). RESUMENObjetivo. La digestibilidad del dsRNA específico para el virus de la mancha blanca (WSSV) por acción de las enzimas del tracto digestivo del camarón Litopenaeus vannamei fue analizada in vitro. Material y métodos. Se midió la actividad de enzimas digestivas (proteasa, amilasa, lipasa, ADNasa y ARNasa) en el estómago, la glándula digestiva, el intestino anterior, medio y posterior en juveniles de camarón patiblanco y se evaluó la digestibilidad de ácidos nucleicos ADN, ARN y dsRNA-ORF89 especifico contra el virus WSSV, por análisis electroforéticos y densitometría. Resultados. La actividad enzimática más alta se encontró en la glándula digestiva del camarón: amilasa (81.41%), lipasa (92.60%), proteasa (78.20%), ADNasa (90.85%) y ARNasa (93.14%). Se evidenció la capacidad digestiva del camarón patiblanco contra el ADN, ARN y dsRNA-ORF89 encontrando en la glándula digestiva la mayor digestión (5.11 ng de ADN por minuto, 8.55 ng de ARN por minuto y 1.48 ng de dsRNA por minuto). Conclusiones. La mayor digestibilidad del dsRNA-ORF89, específico contra el virus WSSV, se encontró en la glándula digestiva y la menor en el intestino posterior. Este es el primer informe relacionado con la digestibilidad del dsRNA-ORF89 por las enzimas del camarón patiblanco con potencial importancia terapéutica en el cultivo de camarón para prevenir la enfermedad del WSSV a través del alimento balanceado. INTRODUCCIÓNUna de las herramientas más poderosas utilizadas en la acuicultura frente a patógenos virales como el virus de la mancha blanca (WSSV) es el mecanismo de ARN de interferencia (ARNi) inducido por ARN de doble cadena (dsRNA, por sus siglas en ingles double-stranded RNA) (1). P...

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Section: Discussionsupporting

confidence: 65%

Section: Discussionunclassified

Section: Discussionmentioning

confidence: 99%

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Digestibilidad in vitro del dsRNA específico por enzimas del tracto digestivo del camarón Litopenaeus vannamei

2017

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“…The immune inductions of shrimp through vaccination techniques have been successfully conducted using protein recombinant WSSV on P. chinensis (Kim et al, 2004) and antiviral dsRNA on L. vannamei (Robalino et al, 2004), and to stimulate the gonad maturation in tiger shrimp (Wulandari, 2010;Treerattrakool et al, 2013), to protect against infectious virus on L. vannamei (Loy et al, 2012;Solis-Lucero et al, 2016),P. japonicas (Namikoshi et al, 2004), and P. monodon (Witteveldt et al, 2004).…”

Section: Characteristic Of a Viral Protein (Vp-15) Of White Spot Syndmentioning

confidence: 99%

CHARACTERISTICS OF VIRAL PROTEIN, VP-15, OF WHITESPOT SYNDROME VIRUS ISOLATED FROM INFECTED TIGER SHRIMP Penaeus monodon (Fabricius, 1798)

Parenrengi

Alimuddin

Tenriulo

2017

Indonesian Aquaculture Journal

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White spot syndrome virus (WSSV) has caused mass mortality on tiger shrimp (Penaeus monodon) culture and adversely affects prawn industry worldwide including Indonesia. It is well known that the protein structure of WSSV plays an important role in the virus infection and morphogenesis process. A viral protein structure called VP-15 is located in the nucleocapsid of virion virus. The protein structure involves in the life cycle of WSSV in host cells. A gene encoding VP-15 could be involved in constructing the RNA interference (RNAi), so it is needed to isolate and characterize for RNAi technology purpose. The study was aimed to isolate and characterize the VP-15 from the infected WSSV tiger shrimp. The characterization of VP-15 was undertaken through assessment of nucleotide sequence, amino acid deduction, alignment nucleotide/protein searches using Genetyx and BLAST program, and dendrogram construction analysis. The results showed that VP-15 was successfully isolated in form of ORFDNA with a fragment size of 243 bp. The phylogenetic tree analysis revealed three clusters corresponding to the time (year) of isolates collection. The VP-15 consisted of 80 amino acids, two start codons (ATG), one stop codon (TAA), and one Kozak context (AAAATGG). Hydrophilic amino acid was the highest composition (44.2%), followed by neutral (31.2%) and hydrophobic (24.6%) amino acid groups. The VP-15 was rich in amino acid of lysine (21.3%), arginine (22.9%) and serine (24.6%). The successful isolation of VP-15 is a very important step in providing a basic yet suitable material in constructing the dsRNA vaccine to control shrimp diseases in aquaculture.

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“…The rHoa-VIH amide however showed very low vitellogenesis-inhibiting activity compared to the natural sinus gland extracts from Marsupenaeus japonicus, which may be due to the use of a heterologous bioassay or the presence of another VIH 20 . In recent studies, silencing of GIH expression in Penaeus monodon using GIH-specific double stranded RNA (GIH-dsRNA) increased the expression level of vitellogenin 21,22 . In a subsequent study, single injection of GIH-dsRNA into previtellogenic female P. monodon led to ovarian maturation and spawning in some prawn indicating that this method was still less effective than unilateral eyestalk ablation 23 .…”

Section: Introductionmentioning

confidence: 99%

Using vitellin monoclonal antibodies to assess the vitellogenesis-inhibiting hormone activity of Macrobrachium rosenbergii

Sripiromrak¹,

Chaivisuthangkura²,

Longyant³

et al. 2014

ScienceAsia

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Vitellogenesis in crustaceans is under the inhibitory regulation of the endocrine system in the X-organ/sinus gland complex in the eyestalks. In this study, two monoclonal antibodies specific to Macrobrachium rosenbergii vitellin and vitellogenin were generated to assess vitellogenesis-inhibiting hormone (VIH) activity. A combination of two monoclonal antibodies specific to the 89 and 100 kDa subunits of vitellin were used to develop a competitive enzyme-linked immunosorbent assay protocol for the quantitative determination of vitellogenin levels in haemolymph. Injection of eyestalk extracts from M. rosenbergii dissolved in either saline or in methanol-acetic acid caused a reduction in vitellogenin levels in the haemolymph at 12 and 24 h post-injection in bilateral eye-ablated prawns with developing ovaries. In contrast, the application of the heterologous eyestalk extract from Penaeus vannamei did not reduce vitellogenin levels in haemolymph. This assay is therefore specific and could be used to assess VIH activity during hormone purification.

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Silencing of gonad-inhibiting hormone gene expression in Penaeus monodon by feeding with GIH dsRNA-enriched Artemia

Cited by 36 publications

References 17 publications

Digestibilidad in vitro del dsRNA específico por enzimas del tracto digestivo del camarón Litopenaeus vannamei

Digestibilidad in vitro del dsRNA específico por enzimas del tracto digestivo del camarón Litopenaeus vannamei

CHARACTERISTICS OF VIRAL PROTEIN, VP-15, OF WHITESPOT SYNDROME VIRUS ISOLATED FROM INFECTED TIGER SHRIMP Penaeus monodon (Fabricius, 1798)

Using vitellin monoclonal antibodies to assess the vitellogenesis-inhibiting hormone activity of Macrobrachium rosenbergii

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