Muhammad Khaliim Jati Kusala, Arif Nur Muhammad Ansori, Reviany V. Nidom, Setyarina Indrasari, Anis F. Astutik, Irine Normalina, Mohammad Y. Alamudi, Siti Rukmana, Kadek Rachmawati, Kuncoro P. Santoso, Chairul A. Nidom
Muhammad Khaliim Jati Kusala1,2, Arif Nur Muhammad Ansori1,2, Reviany V. Nidom2,3, Setyarina Indrasari2,3, Anis F. Astutik2, Irine Normalina2, Mohammad Y. Alamudi2, Siti Rukmana2, Kadek Rachmawati2,4, Kuncoro P. Santoso2,4, Chairul A. Nidom2,4*
1Doctoral Program in Veterinary Science, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
2Professor Nidom Foundation, Surabaya, 60115, Indonesia.
3PT. Riset AIRC Indonesia, Surabaya, 60115, Indonesia.
4Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
Volume - 13,
Issue - 12,
Year - 2020
In recent decades, zebrafish (Danio rerio) has been widely used as a model of biological experiments not only in the immunology and oncology aspects but also experimental animals for infectious diseases in human and fishes. The virus used in this study was the H5N1 avian influenza reverse genetic virus obtained from the Professor Nidom Foundation, Surabaya, Indonesia. We revealed that TCID50 was 3.1623×107/mL, then the virus was developed by inoculating into the primary cell culture and we performed HA test. Samples that showed positive results on the HA test then performed RNA extraction and confirmed by one-step RT-PCR. It can be seen from the presence of cytopathogenic effects (CPE) in primary cell cultures and also can be seen from HA titers in viral harvest samples from the primary cells of zebrafish (Danio rerio) that shows the same result of HA titers is 28. In conclusion, the present study demonstrated that there is an alternative development of new primary cell sources which leads to halal aspects of the production of avian influenza H5N1 vaccines.
Cite this article:
Muhammad Khaliim Jati Kusala, Arif Nur Muhammad Ansori, Reviany V. Nidom, Setyarina Indrasari, Anis F. Astutik, Irine Normalina, Mohammad Y. Alamudi, Siti Rukmana, Kadek Rachmawati, Kuncoro P. Santoso, Chairul A. Nidom. Primary Cell Culture of Zebrafish (Danio rerio) as a material for developing H5N1 Avian Influenza Vaccines. Research J. Pharm. and Tech. 2020; 13(12):6140-6146. doi: 10.5958/0974-360X.2020.01071.9
Muhammad Khaliim Jati Kusala, Arif Nur Muhammad Ansori, Reviany V. Nidom, Setyarina Indrasari, Anis F. Astutik, Irine Normalina, Mohammad Y. Alamudi, Siti Rukmana, Kadek Rachmawati, Kuncoro P. Santoso, Chairul A. Nidom. Primary Cell Culture of Zebrafish (Danio rerio) as a material for developing H5N1 Avian Influenza Vaccines. Research J. Pharm. and Tech. 2020; 13(12):6140-6146. doi: 10.5958/0974-360X.2020.01071.9 Available on: https://rjptonline.org/AbstractView.aspx?PID=2020-13-12-86
1. Artois J, Ippoliti C, Conte A, Dhingra MS, Alfonso P, Tahawy AE, Elbestawy A, Ellakany HF, Gilbert M. Avian influenza A (H5N1) outbreaks in different poultry farm types in Egypt: the effect of vaccination, closing status and farm size. BMC Vet Res. 2018; 14(1): 187.
2. McIntosh EDG. Healthcare-associated infections: potential for prevention through vaccination. Ther Adv Vaccines Immunother. 2018; 6(1): 19-27.
3. Skowronski DM, Janjua NZ, De Serres G, Sabaiduc S, Eshaghi A, Dickinson JA, Fonseca K, Winter AL, Gubbay JB, Krajden M, Petric M, Charest H, Bastien N, Kwindt TL, Mahmud SM, Van Caeseele P, Li Y. Low 2012-13 influenza vaccine effectiveness associated with mutation in the egg-adapted H3N2 vaccine strain not antigenic drift in circulating viruses. PLoS One. 2014; 9(3): e92153.
4. Bardiya N, Bae JH. Influenza vaccines: recent advances in production technologies. Appl Microbiol Biotechnol. 2015; 67(3): 299-305.
5. Eissa AE, Hussein HA, Zaki MM. Detection of avian influenza (H5N1) in some fish and shellfish from different aquatic habitats across some Egyptian Provinces. Life Sci J. 2012; 9(3): 2702-2712.
6. Gabor KA, Goody MF, Mowel WK, Breitbach ME, Gratacap RL, Witten PE, Kim CH. Influenza A virus infection in zebrafish recapitulates mammalian infection and sensitivity to anti-influenza drug treatment. Dis Model Mech. 2014; 7(11): 1227-37.
7. WHO. Cumulative Number of Confirmed Human Cases of Avian Influenza A (H5N1) Reported to WHO, 2003–2017; 2017.
8. Kroehne V, Tsata V, Marrone L, Froeb C, Reinhardt S, Gompf A, Dahl A, Sterneckert J, Reimer MM. Primary spinal OPC culture system from adult zebrafish to study oligodendrocyte differentiation in vitro. Front Cell Neurosci. 2017; 11: 284.
9. Torres-Velarde J, Bautista-Guerrero E, Sifuentes-Romero I, García-Gasca T, García-Gasca A. A muscle-tissue culture system to study myostatin function in fish. Novinka, USA; 2016.
10. Nathiga NKS, Abdul MS, Taju G, Sivasubbu S, Sarath Babu V, Sahul HAS. Effects of nicotine on zebrafish: A comparative response between a newly established gill cell line and whole gills. Comp Biochem Physiol C Toxicol Pharmacol. 2017; 195: 68-77.
11. Meade ME, Roginsky JE, Schulz JR. Primary cell culture of adult zebrafish spinal neurons for electrophysiological studies. J Neurosci Methods. 2019; 322: 50-57.
12. Nidom RV, Alamudi MY, Sillehu S, Indrasari S, Suindarti RD, Qurnianingsih E, Dachlan YP, Aryati, Syaharani A, Rachmawati K, Santoso KP, Nidom CA. Construction of Indonesian-strain avian flu virus seed vaccine using low pathogenic hemagglutinin gene and neuraminidase PR8 gene through reverse genetics. J Vaccines Immunol. 2017; 3(1): 005-011.
13. Poland GA. Vaccines against Avian Influenza—A Race against Time. N Engl J Med. 2006; 354: 1411-1413.
14. Engelhardt OG. Many ways to make an influenza virus - review of influenza virus reverse genetics methods. Influenza Other Respir Viruses. 2013; 7(3): 249-256.
15. Nidom RV, Indrasari S, Lahay AF, Apritasari U, Santoso KP, Nidom CA. H5N1 avian influenza virus infection in zebra fish (Danio rerio) as an alternative media for seed vaccine virus propagation. AIRC Laboratory-Profesor Nidom Foundation, Faculty of Veterinary Medicine, Faculty of Fisheries and Marine, Universitas Airlangga, Surabaya, Indonesia; 2018.
16. LaBarre DD, Lowy RJ. Improvements in methods for calculating virus titer estimates from TCID50 and plaque assays. J Virol Methods. 2001; 96(2): 107-26.
17. Hewajuli DA, Dharrmayanti NLPI. Characterisation and identification of avian influenza virus (AI). Wartazoa. 2008; 18(2).