Infection of Infectious Spleen and Kidney Necrosis Virus and Viral Nervouse Necrosis in Seawater Fish Identifed Using Multiplex Polymerase Chain Reaction
Hidayati Kumalasari(1), Putu Eka Sudaryatma(2*), Artanti Tri Lestari(3), Wahyu Nurlita(4), Wahyu Andy Nugraha(5), Nur Hasanah(6), Ida Ayu Mirah Meliana Dewi(7), Ni Putu Arya Shintya Anggraeni(8)
(1) Program Studi Ilmu Kelautan, Fakultas Pertanian, Universitas Trunojoyo, Madura
(2) Balai Karantina Ikan, Pengendalian Mutu dan Keamanan Hasil Perikanan Denpasar, Kementerian Kelautan dan Perikanan
(3) Balai Karantina Ikan, Pengendalian Mutu dan Keamanan Hasil Perikanan Denpasar, Kementerian Kelautan dan Perikanan
(4) Balai Karantina Ikan, Pengendalian Mutu dan Keamanan Hasil Perikanan Denpasar, Kementerian Kelautan dan Perikanan
(5) Program Studi Ilmu Kelautan, Fakultas Pertanian, Universitas Trunojoyo, Madura
(6) Program Studi Ilmu Kelautan, Fakultas Pertanian, Universitas Trunojoyo, Madura
(7) Program Studi Biologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Udayana, Bali
(8) Program Studi Biologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Udayana, Bali
(*) Corresponding Author
Abstract
Indonesia become a maritime country with large of ocean area and beach line with marine species diversity reaches 37% of the world’s fish. This makes Indonesia have aquaculture potential that supported by an appropriate climate. However, the challenge of viral diseases caused by Infectious Spleen and Kidney Necrosis Virus (ISKNV) and Viral Nervous Necrosis (VNN) would hamper marine aquaculture and causing huge economic losses. Therefore, it is necessary to carry out early detection methods that are efficient, fast, precise and accurate to identifying these viral disease. Here we developed a multiplex-PCR (mPCR) a method that can detect simultaneously of the ISKNV and VNN. The results of this method showed high sensitivity and specificity by using MCP and CP gene target primers to detect ISKNV and VNN, respectively. From a total of 353 samples of seawater fish examined by mPCR, positive results following of single infection of ISKNV and VNN were 14 and 2, respectively. We also found that 18 grouper fish was co-infection with these viruses. From these results, it shows that the mPCR method developed has efficiency, faster and high accuracy. This founding was possible to be applied in laboratory testing or early detection system in the field for ISKNV and VNN.
Keywords
Full Text:
PDFReferences
Angeline, L., Putra, G., Yonathan, C. J., Imania, N., Hilmy, M., Firdaus, R., & Yoewono, J. R. (2020). Stannum : Jurnal Sains dan Terapan Kimia A Review of the Development of Polymerase Chain Reaction Technique and Its Uses in Scientific Field. 2(1), 17–30. https://doi.org/10.33019/jstk.v Ardiansyah, F., Laiila, H., Muharni, M., & Windusari, Y. (2018). Analisis Polimorfisme Padi Varietas Lokal Sumatera Selatan. 7(1), 50–58. Dalla Valle, L., Negrisolo, E., Patarnello, P., Zanella, L., Maltese, C., Bovo, G., & Colombo, L. (2001). Sequence comparison and phylogenetic analysis of fish nodaviruses based on the coat protein gene. Archives of Virology, 146(6), 1125–1137. https://doi.org/10.1007/s007050170110 Dalla Valle, L., Toffolo, V., Lamprecht, M., Maltese, C., Bovo, G., Belvedere, P., & Colombo, L. (2005). Development of a sensitive and quantitative diagnostic assay for fish nervous necrosis virus based on two-target real-time PCR. Veterinary Microbiology, 110(3–4), 167–179. https://doi.org/10.1016/j.vetmic.2005.07.014 Deng, X., Zhang, J., Su, J., Liu, H., Cong, Y., Zhang, L., Zhang, K., & Shi, N. (2018). A multiplex PCR method for the simultaneous detection of three viruses associated with canine viral enteric infections. Archives of Virology, 0123456789, 1–6. https://doi.org/10.1007/s00705-018-3828-4 Go, J., & Whittington, R. (2006). Experimental transmission and virulence of a megalocytivirus (Family Iridoviridae) of dwarf gourami (Colisa lalia) from Asia in Murray cod (Maccullochella peelii peelii) in Australia. Aquaculture, 258(1–4), 140–149. https://doi.org/10.1016/j.aquaculture.2006.04.033 Gustavo, J., Paley, R. R. K., Hunt, W., Ziddah, P. A., Nkansa, M., Guilder, J., Gray, J., & Duodu, S. (2020). First detection of infectious spleen and kidney necrosis virus ( ISKNV ) associated with massive mortalities in farmed tilapia in Africa. Transboundary and Emerging Deseases, 1–14(September). https://doi.org/10.1111/tbed.13825 Heckman, J. J., Pinto, R., & Savelyev, P. A. (2020). Rencana Strategis 2020-2024 Deputi Bidang Koordinasi Sumber Daya Maritim. Angewandte Chemie International Edition, 6(11), 951–952. Inouye, K., Yamano, K., Maeno, Y., Nakajima, K., Matsuoka, M., Wada, Y., & Sorimachi, M. (1992). Iridovirus Infection of Cultured Red Sea Bream, Pagrus major. Fish Pathology, 27(1), 19–27. https://doi.org/10.3147/jsfp.27.19 Khoironi, F. E., & Saskara, I. A. N. (2015). Analisis Pengaruh Kurs Dollar, Inflasi, dan Produksi terhadap Ekspor Ikan Hias di Provinsi Bali. E-Journal EP Unud, 6(3), 337–361. Kim, H., Ryu, J., Lee, S., Kim, E., & Kim, H. (2015). Multiplex PCR for detection of the Vibrio genus and five pathogenic Vibrio species with primer sets designed using comparative genomics. BMC Microbiology, 1–11. https://doi.org/10.1186/s12866-015-0577-3 Kurita, J., & Nakajima, K. (2012). Megalocytiviruses. Viruses, 4(4), 521–538. https://doi.org/10.3390/v4040521 Langga, I. F., Restu, M., & Kuswinanti, T. (2012). Optimalisasi suhu dan lama inkubasi dalam ekstraksi dna tanaman bitti (. J Sains & Teknologi, 12(3), 265–276. Lorenz, T. C. (2012). Polymerase Chain Reaction : Basic Protocol Plus Troubleshooting and Optimization Strategies. May, 1–14. https://doi.org/10.3791/3998 Ma, H., Xie, J., Weng, S., Zhou, T., & He, J. (2012). Co-infection of megalocytivirus and viral nervous necrosis virus in a very severe mass mortality of juvenile orange-spotted groupers ( Epinephelus coioides ). Aquaculture, 358–359, 170–175. https://doi.org/10.1016/j.aquaculture.2012.06.032 Maksum, I. P., Amalia, R., Kamara, D. S., Diana, S., & Rachman, R. W. (2018). PCR Multipleks untuk Identifikasi Mycobacterium tuberculosis Resisten terhadap Isoniazid dan Rifampisin pada Galur Lokal Balai Laboratorium Kesehatan Provinsi Jawa Barat Multiplex PCR for Identification the Isoniazid- and Rifampin-resistant Mycobacterium tuberculosis ( Local Strain of Balai Laboratorium Kesehatan Provinsi Jawa Barat ). 4(November), 107–118. Murwantoko, Handayani, christina retna, & Pratiwi, R. (2009). Cloning and Sequence Analysis of Capsid Protein Gene of Iridovirus Indonesian Isolates. Indonesian Journal of Biotechnology, 14(1), 1117–1123. Nguyen, H. D., Mekuchi, T., Imura, K., Nakai, T., Nishizawa, T., & Muroga, K. (1994). Occurrence of Viral Nervous Necrosis (VNN) in Hatchery-Reared Juvenile Japanese Flounder Paralichthys olivaceus. Fisheries Science, 60(5), 551–554. https://doi.org/10.2331/fishsci.60.551 Nishizawa, T., Mori -i., K., Furuhashi, M., Nakai, T., Furusawa, I., & Muroga, K. (1995). Comparison of the coat protein genes of five fish nodaviruses, the causative agents of viral nervous necrosis in marine fish. Journal of General Virology, 76(7), 1563–1569. https://doi.org/10.1099/0022-1317-76-7-1563 Puspasari, K., W, Z., Nurdin, A., Hasriani, Freddy, R., & Ishaaq, S. (2020). Pemetaan Genomik Daerah RNA2 (Coat Protein) Betanodavirus Penyebab Penyakit Viral Nervous Necrosis (VNN). Jurnal Kelautan Dan Perikanan Terapan, 3(2), 79–87. Putri, D. A. S., Rosjadi, F., Sundari, M. S., & DAYA. (2016). Dwi Ayu Sekarini Putri, Firman Rosjadi, Made Siti Sundari. Ekonomi Dan Bisnis, 2017, 11–18. Rifai, A. B., Luh, N., Ika, P., Yulianah, L., Pasaribu, F. H., Medik, D. M., Ilmu, D., Hewan, P., Karantina, S., Pengendalian, I., & Riau, K. (2020). Infeksi Megalocytivirus pada Budidaya Ikan Air Laut dan Ikan Air Tawar di Beberapa Provinsi di Indonesia. 21(36), 423–434. https://doi.org/10.19087/jveteriner.2020.21.3.423 Rimmer, A. E., Becker, J. A., Tweedie, A., & Whittington, R. J. (2012). Development of a quantitative polymerase chain reaction (qPCR) assay for the detection of dwarf gourami iridovirus (DGIV) and other megalocytiviruses and comparison with the Office International des Epizooties (OIE) reference PCR protocol. Aquaculture, 358–359, 155–163. https://doi.org/10.1016/j.aquaculture.2012.06.034 Sarjito, I., Ir, P., Budi, S., Harjuno, A., & Haditomo, C. (2013). PARASIT DAN PENYAKIT IKAN. Sembiring, S. B. M., Wibawa, G. S., Mahardika, K., Widiastuti, Z., & Haryanti, H. (2018). Prevalensi Infeksi Viral Nervous Necrosis (Vnn) Dan Iridovirus Pada Hatcheri Dan Budidaya Ikan Laut. Media Akuakultur, 13(2), 83. https://doi.org/10.15578/ma.13.2.2018.83-90 Sudaryatma, P. E., Lestari, A. T., Sunarsih, N. L., Widiarti, K. S., Hidayah, S. N., & Srinoto, D. (2012). Imunositokimia Streptavidin Biotin : Deteksi Dini Viral Nervous Necrosis Virus pada Lendir Ikan Kerapu Macan ( Epinephelus fuscoguttatus) [in Indonesian]. Jurnal Sain Veteriner, 30(March), 99–109. Thamizhvanan, S., Sivakumar, S., Santhosh Kumar, S., Vinoth Kumar, D., Suryakodi, S., Balaji, K., Rajkumar, T., Vimal, S., Abdul Majeed, S., Taju, G., & Sahul Hameed, A. S. (2019). Multiple infections caused by white spot syndrome virus and Enterocytozoon hepatopenaei in pond-reared Penaeus vannamei in India and multiplex PCR for their simultaneous detection. Journal of Fish Diseases, 42(3), 447–454. https://doi.org/10.1111/jfd.12956 Toffan, A., Pascoli, F., Pretto, T., Panzarin, V., Abbadi, M., Buratin, A., Quartesan, R., Gijón, D., & Padrós, F. (2017). Viral nervous necrosis in gilthead sea bream ( Sparus aurata ) caused by reassortant betanodavirus RGNNV / SJNNV : an emerging threat for Mediterranean aquaculture. May, 1–12. https://doi.org/10.1038/srep46755 Toubanaki, D. K., Efstathiou, A., & Karagouni, E. (2022). Transcriptomic Analysis of Fish Hosts Responses to Nervous Necrosis Virus. Pathogens, 11 (201), 1–30. Uslan, U., & Pharmawati, M. (2015). Optimasi Konsentrasi DNA dan MgCl2 pada Reaksi Polymerase Chain Reaction-Random Amplified Polymorphic DNA untuk Analisis Keragaman Genetik Tanaman Faloak (Sterculia quadrifida R.Br) (Optimization of DNA and MgCl2 Concentrations in Polymerase Chain Reacti. Jurnal Bios Logos, 5(1). https://doi.org/10.35799/jbl.5.1.2015.9316 Yang, Z., Yue, G. H., & Wong, S. M. (2021). VNN disease and status of breeding for resistance to NNV in aquaculture. Aquaculture and Fisheries, April. https://doi.org/10.1016/j.aaf.2021.04.001 Yoshikoshi, K., & Inoue, K. (1990). Viral nervous necrosis in hatchery‐reared larvae and juveniles of Japanese parrotfish, Oplegnathus fasciatus (Temminck & Schlegel). Journal of Fish Diseases, 13(1), 69–77. https://doi.org/10.1111/j.1365-2761.1990.tb00758.x Yuenleni. (2019). LANGKAH-LANGKAH OPTIMASI PCR ISSN 2655 4887 ( Print ), ISSN 2655 1624 ( Online ) ISSN 2655 4887 ( Print ), ISSN 2655 1624 ( Online ). 1(3), 51–56. Zafran, Koesharyani, I., Johnny, F., Yuasa, K., Harada, T., & Hatai, K. (2012). Viral Nervous Grouper altivelis Necrosis Larvae in in Hump and Cromileptes Juveniles Indonesia. Fish Pathology, 35(2), 95–96. Zafran, Z., Harada, T., I. Koesharyan, I. K., Yuasa, K., & Hatai, K. (1998). Indonesian Hatcery Reared Seabass Larvae (Lates calcarifer), Associated With Viral Nervous Necrosis (VNN). Indonesian Fisheries Research Journal, 4(1), 19. https://doi.org/10.15578/ifrj.4.1.1998.19-22
DOI: https://doi.org/10.22146/jsv.73599
Article Metrics
Abstract views : 953 | views : 2569Refbacks
- There are currently no refbacks.
Copyright (c) 2022 Jurnal Sain Veteriner
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Jurnal Sain Veteriner Indexed by
Copyright of JSV (Jurnal Sain Veteriner) ISSN 0126-0421 (print), ISSN 2407-3733 (online).
Fakultas Kedokteran Hewan, Universitas Gadjah Mada
Jl. Fauna No.2, Karangmalang, Yogyakarta
Phone: 0274-560862
Fax: 0274-560861
Email: jsv_fkh@ugm.ac.id
HP. 0895363078367
Jurnal Sain Veteriner is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
View My Stats