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RISS 인기검색어
- 검색키워드
- 저자 : Sajal Kole (검색결과 3 건)
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Kole, Sajal,Qadiri, Syed Shariq Nazir,Shin, Su-Mi,Kim, Wi-Sik,Lee, Jehee,Jung, Sung-Ju ACADEMIC PRESS LTD 2019 FISH AND SHELLFISH IMMUNOLOGY Vol.91 No.-
<P><B>Abstract</B></P> <P>Viral haemorrhagic septicaemia virus (VHSV), a (−) ssRNA virus belonging to the genus <I>Novirhabdovirus</I> of rhabdoviridae family, is the aetiological agent of viral haemorrhagic septicaemia (VHS) disease which causes huge economic losses in farmed olive flounder (<I>Paralichthys olivaceus</I>) and significant mortalities among several other marine fish species in Korea, Japan, and China. Previously, we developed an inactivated vaccine viz., formalin-inactivated VHSV mixed with squalene as adjuvant which was effective in conferring protective immunity (58–76% relative percentage survival) against VHSV but the mode of administration was intraperitoneal injection which is not feasible for small sized fingerling fish. To overcome this limitation, we presently focused on replacing the injection route of vaccine delivery by oral and immersion routes. In this context, we encapsulated the inactivated VHSV vaccine with chitosan nanoparticles (CNPs-IV) by water-in-oil (W/O) emulsification method. After encapsulation, two sets of <I>in vivo</I> vaccination trials were conducted viz., preliminary trial-I and final trial-II. In preliminary trial-I, olive flounder fingerlings (10.5 ± 1.7 g) were vaccinated with CNPs-IV by different delivery strategies involving oral and immersion routes (single/booster dose) followed by challenge with VHSV (1 × 10<SUP>6</SUP> TCID<SUB>50</SUB> virus/fish) to evaluate an effective method amongst different applied delivery strategies. Subsequently, a final trial-II was conducted to better understand the immune mechanism behind the efficacy of the employed delivery strategy and also to further improvise the delivery mechanism with prime-boost (primary immersion and oral boosting) combination in order to improve the transient anti-VHSV response in the host. Evaluation of RPS analysis in trial-I revealed higher RPS of 46.7% and 53.3% in the CNPs-IV (immersion) and CNPs-IV (immersion/immersion) groups, respectively compared to 0% RPS in the CNPs-IV (oral) group and 20% RPS in the CNPs-IV (oral/oral) group when calculated against 100% cumulative mortality percentage in the NVC (non-vaccinated challenged) control group, whereas, in the trial-II, RPS of 60% and 66.6% were obtained for CNPs-IV (immersion/immersion) and CNPs-IV (immersion/oral) groups, respectively. In addition, specific (anti-VHSV) antibody titre in the fish sera, skin mucus and intestinal mucus of the immunized groups were significantly (p < 0.05) enhanced following vaccination. Furthermore, CNPs-IV immunized fish showed significant (p < 0.05) upregulation of different immune gene transcripts (IgM, IgT, pIgR, MHC-I, MHC-II, IFN-γ, and Caspase3) compared to control, in both the systemic (kidney) and mucosal (skin and intestine) immune compartments of the host post immunization as well as post challenge. To conclude, mucosal immunization with CNPs-IV vaccine can orchestrate an effective immunization strategy in organizing a coordinative immune response against VHSV in olive flounder thereby exhibiting higher protective efficacy to the host with minimum stress.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Encapsulation of inactivated VHSV in chitosan NPs for preparation of CNPs-IV vaccine. </LI> <LI> Olive flounder fingerlings immunized with CNPs-IV through mucosal routes. </LI> <LI> CNPs-IV vaccinated fish exhibited high RPS (60–66.6%) post VHSV challenged. </LI> <LI> CNPs-IV immunized group showed higher specific antibody response in sera and mucus. </LI> <LI> Immune genes were significantly upregulated in immunized fish. </LI> </UL> </P>
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Qadiri, Syed Shariq Nazir,Kim, Soo-Jin,Krishnan, Rahul,Kim, Jae-Ok,Kole, Sajal,Kim, Wi-Sik,Oh, Myung-Joo Elsevier 2019 Aquaculture Vol.505 No.-
<P><B>Abstract</B></P> <P>An insight on localization of viral pathogen is imperative to better understand the host-pathogen interaction. Previously, we developed an <I>in-situ</I> hybridization (RNA-ISH) assay to detect viral hemorrhagic septicemia virus (VHSV), an OIE reportable fish pathogen, in an <I>in vitro</I> model. Here, we utilized its potential and applicability <I>in vivo</I> to further our understanding about the localization and tissue tropism of VHSV in experimentally infected juvenile olive flounder (<I>Paralichthys olivaceus</I>), an economically important flatfish in Asian aquaculture. Two separate digoxigenin (DIG) labeled antisense RNA probes targeting a fragment of viral nucleoprotein (N) and glycoprotein (G) gene segments were employed to localize VHSV in five infected flounder tissues <I>viz.</I>, kidney, spleen, heart, liver and brain. Further, immunohistochemistry (IHC) assay was also used to observe and substantiate the spatial localization of the viral particles using a monoclonal antibody (MAb) against the viral nucleoprotein (N). Following the RNA-ISH and IHC assays, VHSV localization was observed in different cells and areas of the tested tissues. The positive reaction in both the assays correlated with histopathological alterations confirming the presence of VHSV within the tested tissues. Information gained from the present study using both genome and protein based localization sheds light on viral tropism which can help to further elucidate VHSV pathogenesis in olive flounder.</P> <P><B>Highlights</B></P> <P> <UL> <LI> RNA-ISH assay was used for the first time to localize VHSV in olive flounder tissues. </LI> <LI> Viral mRNAs were visualized in different areas of tested tissues using two DIG-labeled riboprobes (VHSV-N and G). </LI> <LI> Protein based detection (IHC) and histopathological alterations also confirmed the viral localization. </LI> <LI> The present study sheds light on VHSV tissue tropism and pathogenesis in olive flounder. </LI> </UL> </P>
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