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Amal, Mohammad Noor Azmai
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Biochemistry, Genetics & Molecular Biology

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Stability and Efficacy of Live-Attenuated Vibrio harveyi Vaccines Under Different Storage Conditions in Zebrafish (Danio rerio) Models Muhamad-Sofie, Mohd Hafiz Ngoo; Mohamad, Aslah; Azzam-Sayuti, Mohamad; Amal, Mohammad Noor Azmai; Zamri-Saad, Mohd; Monir, Md Shirajum; Md Yasin, Ina Salwany
Jurnal Ilmiah Perikanan dan Kelautan Vol. 16 No. 2 (2024): JURNAL ILMIAH PERIKANAN DAN KELAUTAN
Publisher : Faculty of Fisheries and Marine Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/jipk.v16i2.59794

Abstract

Graphical Abstract Highlight Research The LD50 (median lethal dose) of Vibrio parahaemolyticus and harveyi in zebrafish was determined to be 1 x 106 CFU/mL, while for V. alginolyticus it was found to be 1 x 105 CFU/mL. The LAVh vaccine demonstrated cross-protection against various pathogenic strains of Vibrio, leading to an average of 80% survival rate in vaccinated individuals. The analysis of the LAVh vaccine emphasized its versatility, as it can be quickly deployed and stored as a freeze-dried powder. The LAVh vaccine can be easily accessible and user-friendly in various aquaculture environments, such as offshore and remote farms. Abstract Vibriosis poses a significant threat to marine teleosts, causing substantial losses in the global aquaculture industry. Previous work in our lab led to the development of a live-attenuated V. harveyi vaccine (LAVh) candidate that targets the serine endoprotease gene with a three-point knockout and has shown promise in protecting against vibriosis. However, further investigation is necessary to evaluate the stability and efficacy of its various storage conditions for broader applications. This study aims to determine how well the three different LAVh vaccine storage (fresh, stale, and freeze-dried LAVh) worked against vibriosis. A total of 1000 adult zebrafish (Danio rerio) (mean weight: 0.20±0.5 g) were divided into four groups. Groups 1, 2, and 3 were intraperitoneally injected with different LAVh vaccine storage (fresh, stale, and freeze-dried, respectively), while Group 4 received 0.01 M phosphate-buffered saline (PBS) and served as the unvaccinated control. Fish were monitored for 21 days post-vaccination for safety, stability, efficacy, and antibody analysis. The results showed that a modest dosage of 1 x 104 CFU/mL of LAVh vaccine from all storage conditions provided 80% survival upon intraperitoneal challenge with pathogenic strains of pathogenic V. harveyi, V. alginolyticus, and V. parahaemolyticus. This dosage induced significant antibody production and conferred cross-protection against different Vibrio spp., indicating the LAVh vaccine’s potential for commercial application. The LAVh vaccine demonstrated high effectiveness and suitability for storage as a freeze-dried powder. This study might offer significant insights into practical strategies for reducing vibriosis, especially in aquaculture settings with limited infrastructure.
Designing of a Novel Aerolysin-based Multiepitope Vaccine against Aeromonas hydrophila Isolated from Osphronemus goramy Using Reverse Vaccinology: an in Silico Approaches Rozi; Tyasningsih, Wiwiek; Rahmahani, Jola; Aksono, Eduardus Bimo; Yunus , Muchammad; Al-Arif, Mohammad Anam; Kuncorojati, Suryo; Kusdarwati, Rahayu; Sari, Putri Desi Wulan; Amal, Mohammad Noor Azmai; Salleh, Annas; Khanand, Nadeem; Suwarno
Jurnal Ilmiah Perikanan dan Kelautan Vol. 16 No. 2 (2024): JURNAL ILMIAH PERIKANAN DAN KELAUTAN
Publisher : Faculty of Fisheries and Marine Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/jipk.v16i2.62035

Abstract

Graphical Abstract Highlight Research The study aims to develop a multi-epitope vaccine (MEV) against A. hydrophila by targeting the aerolysin toxin, a key virulence factor responsible for infections in fish and humans. Computational methods identified and optimized B-cell and T-cell epitopes, focusing on their ability to trigger immune responses without causing toxicity or allergenicity. In silico simulations demonstrated that the MEV has a strong binding affinity to immune receptors like TLR-4, MHC-I, and MHC-II, indicating its potential to induce robust cellular and humoral immunity. Structural analysis of the MEV showed a stable 3D conformation, with most residues in favorable regions, ensuring stability during immune activation. The MEV could enhance disease control in aquaculture and reduce human infection risks, offering a promising solution to address antibiotic resistance and the absence of effective vaccines. Abstract Aeromonas hydrophila, gram-negative, is a major pathogen responsible for various diseases in mammals, reptiles, amphibia, and vertebrates, including fish and humans. Targeting the specific toxin aerolysin in A. hydrophila is crucial to address antibiotic resistance and the lack of adequate and protective vaccines against this intracellular pathogen. This study aimed to identify a multi-epitope vaccination (MEV) candidate targeting A. hydrophila aerolysin toxin to combat the disease effectively. Standard biochemical characterization methods and sequencing of the 16S rRNA, rpoB, and aerA genes identified the isolate AHSA1 as A. hydrophila. Subsequently, we identified B and T cell epitopes on the aerolysin protein and separately predicted MHC-I and MHC-II epitopes. The epitopes are then evaluated for toxicity, antigenicity, allergenicity, and solubility. The vaccine design integrated multi-epitope-based constructs, utilizing specialized linkers (GPGPG) and EAAAK linkers to connect epitope peptides with adjuvants in the cholera toxin B component, thereby enhancing immunogenicity. Ramachandran plots showed that 85.25% of the residues were located in the most favorable regions, which was followed by the generously allowed zone (1.30%), the additional allowed regions (10.80%), and the forbidden regions (2.65%), thus confirming the feasibility of the modeled vaccine design. Based on docking simulations, MEV had the highest binding and interaction energies with TLR-4, TLR-9, MHC-I, and MHC-II (-1081.4, -723.2, 866.2, -9043.3 kcal/mol). Based on computational modelling, we expect the Aerolysin MEV candidate design to activate diverse immune mechanisms, stimulate robust responses against A. hydrophila, and maintain safety. The significant solubility, absence of toxicity or allergic response, and minimal side effects in animal testing all contribute to the potential clinical utility of this vaccine candidate.  
Co-Authors Aksono HP., Eduardus Bimo Al arif, Mohammad Anam Azzam-Sayuti, Mohamad Khanand, Nadeem Kuncorojati, Suryo Md Yasin, Ina Salwany Mohamad, Aslah Monir, Md Shirajum Muhamad-Sofie, Mohd Hafiz Ngoo Rahmahani, Jola Rozi Salleh, Annas Sari, Putri Desi Wulan Suwarno Tyasningsih, Wiwiek Yunus , Muchammad Zamri-Saad, Mohd