Aeromonas hydrophila OmpW PLGA nanoparticle oral vaccine shows a dose-dependent protective immunity in rohu (Labeo rohita)

Saurabh Dubey, Kiran Avadhani, Srinivas Mutalik, Sangeetha Madambithara Sivadasan, Biswajit Maiti, Joydeb Paul, Shivani Kallappa Girisha, Moleyur Nagarajappa Venugopal, Stephen Mutoloki, Øystein Evensen, Indrani Karunasagar, Hetron Mweemba Munang’andu

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Aeromonas hydrophila is a Gram-negative bacterium that causes high mortality in different fish species and at different growth stages. Although vaccination has significantly contributed to the decline of disease outbreaks in aquaculture, the use of oral vaccines has lagged behind the injectable vaccines due to lack of proven efficacy, that being from primary immunization or by use of boost protocols. In this study, the outer membrane proteinW(OmpW) of A. hydrophila was cloned, purified, and encapsulated in poly D,L-lactide-co-glycolic acid (PLGA) nanoparticles (NPs) for oral vaccination of rohu (Labeo rohita Hamilton). The physical properties of PLGA NPs encapsulating the recombinant OmpW (rOmpW) was characterized as having a diameter of 370–375 nm, encapsulation efficiency of 53% and -19.3 mV zeta potential. In vitro release of rOmpW was estimated at 34% within 48 h of incubation in phosphate-buffered saline. To evaluate the efficacy of the NP-rOmpW oral vaccine, two antigen doses were orally administered in rohu with a high antigen (HiAg) dose that had twice the amount of antigens compared to the low antigen (LoAg) dose. Antibody levels obtained after vaccination showed an antigen dose dependency in which fish from the HiAg group had higher antibody levels than those from the LoAg group. The antibody levels corresponded with post challenge survival proportions (PCSPs) and relative percent survival (RPS) in which the HiAg group had a higher PCSP and RPS than the LoAg group. Likewise, the ability to inhibit A. hydrophila growth on trypticase soy agar (TSA) by sera obtained from the HiAg group was higher than that from the LoAg group. Overall, data presented here shows that OmpW orally administered using PLGA NPs is protective against A. hydrophila infection with the level of protective immunity induced by oral vaccination being antigen dose-dependent. Future studies should seek to optimize the antigen dose and duration of oral immunization in rohu in order to induce the highest protection in vaccinated fish.

Original languageEnglish
Article number21
JournalVaccines
Volume4
Issue number2
DOIs
Publication statusPublished - 01-06-2016

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glycolic acid
Aeromonas hydrophila
Nanoparticles
Immunity
Vaccines
Antigens
Membranes
Vaccination
Fishes
poly(lactide)
Antibodies
Immunization

All Science Journal Classification (ASJC) codes

  • Immunology
  • Pharmacology
  • Drug Discovery
  • Pharmacology (medical)
  • Infectious Diseases

Cite this

Dubey, Saurabh ; Avadhani, Kiran ; Mutalik, Srinivas ; Sivadasan, Sangeetha Madambithara ; Maiti, Biswajit ; Paul, Joydeb ; Girisha, Shivani Kallappa ; Venugopal, Moleyur Nagarajappa ; Mutoloki, Stephen ; Evensen, Øystein ; Karunasagar, Indrani ; Munang’andu, Hetron Mweemba. / Aeromonas hydrophila OmpW PLGA nanoparticle oral vaccine shows a dose-dependent protective immunity in rohu (Labeo rohita). In: Vaccines. 2016 ; Vol. 4, No. 2.
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title = "Aeromonas hydrophila OmpW PLGA nanoparticle oral vaccine shows a dose-dependent protective immunity in rohu (Labeo rohita)",
abstract = "Aeromonas hydrophila is a Gram-negative bacterium that causes high mortality in different fish species and at different growth stages. Although vaccination has significantly contributed to the decline of disease outbreaks in aquaculture, the use of oral vaccines has lagged behind the injectable vaccines due to lack of proven efficacy, that being from primary immunization or by use of boost protocols. In this study, the outer membrane proteinW(OmpW) of A. hydrophila was cloned, purified, and encapsulated in poly D,L-lactide-co-glycolic acid (PLGA) nanoparticles (NPs) for oral vaccination of rohu (Labeo rohita Hamilton). The physical properties of PLGA NPs encapsulating the recombinant OmpW (rOmpW) was characterized as having a diameter of 370–375 nm, encapsulation efficiency of 53{\%} and -19.3 mV zeta potential. In vitro release of rOmpW was estimated at 34{\%} within 48 h of incubation in phosphate-buffered saline. To evaluate the efficacy of the NP-rOmpW oral vaccine, two antigen doses were orally administered in rohu with a high antigen (HiAg) dose that had twice the amount of antigens compared to the low antigen (LoAg) dose. Antibody levels obtained after vaccination showed an antigen dose dependency in which fish from the HiAg group had higher antibody levels than those from the LoAg group. The antibody levels corresponded with post challenge survival proportions (PCSPs) and relative percent survival (RPS) in which the HiAg group had a higher PCSP and RPS than the LoAg group. Likewise, the ability to inhibit A. hydrophila growth on trypticase soy agar (TSA) by sera obtained from the HiAg group was higher than that from the LoAg group. Overall, data presented here shows that OmpW orally administered using PLGA NPs is protective against A. hydrophila infection with the level of protective immunity induced by oral vaccination being antigen dose-dependent. Future studies should seek to optimize the antigen dose and duration of oral immunization in rohu in order to induce the highest protection in vaccinated fish.",
author = "Saurabh Dubey and Kiran Avadhani and Srinivas Mutalik and Sivadasan, {Sangeetha Madambithara} and Biswajit Maiti and Joydeb Paul and Girisha, {Shivani Kallappa} and Venugopal, {Moleyur Nagarajappa} and Stephen Mutoloki and {\O}ystein Evensen and Indrani Karunasagar and Munang’andu, {Hetron Mweemba}",
year = "2016",
month = "6",
day = "1",
doi = "10.3390/vaccines4020021",
language = "English",
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Dubey, S, Avadhani, K, Mutalik, S, Sivadasan, SM, Maiti, B, Paul, J, Girisha, SK, Venugopal, MN, Mutoloki, S, Evensen, Ø, Karunasagar, I & Munang’andu, HM 2016, 'Aeromonas hydrophila OmpW PLGA nanoparticle oral vaccine shows a dose-dependent protective immunity in rohu (Labeo rohita)', Vaccines, vol. 4, no. 2, 21. https://doi.org/10.3390/vaccines4020021

Aeromonas hydrophila OmpW PLGA nanoparticle oral vaccine shows a dose-dependent protective immunity in rohu (Labeo rohita). / Dubey, Saurabh; Avadhani, Kiran; Mutalik, Srinivas; Sivadasan, Sangeetha Madambithara; Maiti, Biswajit; Paul, Joydeb; Girisha, Shivani Kallappa; Venugopal, Moleyur Nagarajappa; Mutoloki, Stephen; Evensen, Øystein; Karunasagar, Indrani; Munang’andu, Hetron Mweemba.

In: Vaccines, Vol. 4, No. 2, 21, 01.06.2016.

Research output: Contribution to journalArticle

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T1 - Aeromonas hydrophila OmpW PLGA nanoparticle oral vaccine shows a dose-dependent protective immunity in rohu (Labeo rohita)

AU - Dubey, Saurabh

AU - Avadhani, Kiran

AU - Mutalik, Srinivas

AU - Sivadasan, Sangeetha Madambithara

AU - Maiti, Biswajit

AU - Paul, Joydeb

AU - Girisha, Shivani Kallappa

AU - Venugopal, Moleyur Nagarajappa

AU - Mutoloki, Stephen

AU - Evensen, Øystein

AU - Karunasagar, Indrani

AU - Munang’andu, Hetron Mweemba

PY - 2016/6/1

Y1 - 2016/6/1

N2 - Aeromonas hydrophila is a Gram-negative bacterium that causes high mortality in different fish species and at different growth stages. Although vaccination has significantly contributed to the decline of disease outbreaks in aquaculture, the use of oral vaccines has lagged behind the injectable vaccines due to lack of proven efficacy, that being from primary immunization or by use of boost protocols. In this study, the outer membrane proteinW(OmpW) of A. hydrophila was cloned, purified, and encapsulated in poly D,L-lactide-co-glycolic acid (PLGA) nanoparticles (NPs) for oral vaccination of rohu (Labeo rohita Hamilton). The physical properties of PLGA NPs encapsulating the recombinant OmpW (rOmpW) was characterized as having a diameter of 370–375 nm, encapsulation efficiency of 53% and -19.3 mV zeta potential. In vitro release of rOmpW was estimated at 34% within 48 h of incubation in phosphate-buffered saline. To evaluate the efficacy of the NP-rOmpW oral vaccine, two antigen doses were orally administered in rohu with a high antigen (HiAg) dose that had twice the amount of antigens compared to the low antigen (LoAg) dose. Antibody levels obtained after vaccination showed an antigen dose dependency in which fish from the HiAg group had higher antibody levels than those from the LoAg group. The antibody levels corresponded with post challenge survival proportions (PCSPs) and relative percent survival (RPS) in which the HiAg group had a higher PCSP and RPS than the LoAg group. Likewise, the ability to inhibit A. hydrophila growth on trypticase soy agar (TSA) by sera obtained from the HiAg group was higher than that from the LoAg group. Overall, data presented here shows that OmpW orally administered using PLGA NPs is protective against A. hydrophila infection with the level of protective immunity induced by oral vaccination being antigen dose-dependent. Future studies should seek to optimize the antigen dose and duration of oral immunization in rohu in order to induce the highest protection in vaccinated fish.

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