Murwantoko Murwantoko, Endang Wijayanti, Sri Agustatik, Harshelly Valianti, Lalla Kumala Yulanda, Nur Lailatul Firotun Nukmah


The orange clownfish, Amphiprion percula, is a popular fish in the global marine ornamental trade. In March 2021, several orange clown fish at Batam Mariculture Development Center exhibited lethargic behavior with wart-like nodules on the fins suggesting a viral disease infection. This study aimed to detect the suspected viral disease agent from the clownfish and determine its genotype based on the major capsid protein gene. The fish tissues with wart-like nodules were collected and fixed in 96% ethanol. The DNA was extracted from the tissues and used as the template for the amplification of the major capsid protein (MCP) and myristylated membrane protein (MMP) genes using polymerase chain reaction (PCR). The nucleotide sequences of the PCR products were analyzed for their homology using the Basic Local Alignment Search Tool (BLAST). Multiple alignments of the amino acid sequence of MCP were performed using MEGA-X to determine the genotype. The PCR amplification produced the expected bands for detecting MCP, MMP and DNAPol genes. These results indicated the presence of Lymphocystis disease virus (LCDV), designated as LCDV-Oc-Btm. The sequencing of MCP and MMP genes produced the 1221 and 407 nucleotides, respectively. The BLAST analysis showed the highest identity was obtained with the species of LCDV-1 (LCDV-Sa strain) at 91.04 % and 88.19 % for MCP and MMP, respectively. The UPGMA phylogenic tree showed LCDV-Oc-Btm differs from the existing genotype and can be assigned as a new genotype. This study concludes that LCDVOc-Btm is a novel species of lymphocystis disease virus.


DNA polymeras; Genotype; LCDV; MCP; MMP

Full Text:



Cano, I., Valverde, E.J., Lopez, J.B., Alonso, M.C., Garcia, R.E., Sarasquete, C., Borrego, J.J., & Castro, D. (2010). A New Genotype of Lymphocystivirus Isolated from Cultured Gilthead Seabream, Sparus aurata L., and Senegalese sole, Silea senegalensis (Kaup). Journal of Fish Disease. 33 (8), 695-700.;

Chinchar, V.G., Waltzek, T.B., & Subramaniam, K. (2017). Ranaviruses and Other Members of The Family Iridoviridae: Their Place in The Virosphere. Virology, 17, 30184-30188.

Doszpoly, A., Kajan, G., Puentes, R., & Perretta, A. (2020). Complete Genome Sequence and Analysis of a Novel Lymphocystivirus Detected in Whitemouth Croaker (Micropogonias furnieri): Lymphocystis Disease Virus 4. Archives of Virology, 165 (5), 1215 – 1218.

Fu, X.Z., Shi, C.B., Li, N.Q., Chang, O.Q., & Wu, S.Q. (2007). Inverse PCR Amplification of The Complete Major Capsid Protein Gene of Lymphocystis Disease Virus Isolated from Rachycentron canadum and Phylogenetic Analysis of The Virus. Bing Du Xue Bao, 23 (5), 412-416.

Hick, P.M., Becker, J.A., & Whittington, R.J. (2016). Iridovirus of Fish (Chapter 8). In Kibenge, F.S.B., & Godoy, M., USA (Ed.) Aquaculture Virology (pp.127-152). Ist Ed. Amsterdam: Elsevier Inc.

Hossain, M., Song, J.Y., Kitamura, S.I., Jung, S.J., & Oh, M.J. (2008). Phylogenetic Analysis of Lymphocystis Disease Virus from Tropical Ornamental Fish Species Based On A Major Capsid Protein Gene. Journal Of Fish Disease, 31 (6), 473-479.

Huang, X., Huang, Y., Xu, L., Wei, S., Ouyang, Z., Feng, J., & Qin, Q. (2015). Identification and Characterization of a Novel Lymphocyctis Disease Virus Isolate from Cultured Grouper in China. Journal of Fish Disease, 38 (4), 379-387.

Kim, T.J., & Lee, J.I. (2007). Sequence Variation in The Genes Encoding The Major Capsid Protein, The ATPase, and The RNA Polymerase 2 (Domain 6) of Lymphocystis Disease Virus Isolated From Schlegel’s Black Rockfish, Sebastes schlegelii Hilgendorf. Journal of Fish Disease, 30 (8), 501-504.

Kitamura, S.I., Jung, S.J., & Oh, M.J. (2006b). Differentiation of Lymphocystic Disease Virus Genotype by Multiplex PCR. The Journal of Microbiology, 44 (2), 248-253.

Kitamura, S.I., Jung, S.J., Kim, W.S., Nishizawa, T., Yoshimizu, M., & Oh, M.J. (2006). A New Genotype Of Lymphocystivirus, LCDV-RF, From Lymphocystis Diseased Rockfish. Archives of Virology, 151 (3), 607-615.

Kumar, S., Stecher, G., & Tamura, K. (2016). Mega7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular Biology and Evolution, 33 (7), 1870-1874.

Lam, C., Khairunissa., Damayanti, L., Kurobe, T., Teh, S.J., Pfahl, H., Rapi, S., Janetski, N., & Baxa, D.V. (2020). Detection of New Strain of Lymphocystis Disease Virus (LCDV) in Captive-Bred Clownfish Amphiprion percula in South Sulawesi, Indonesia. Aquaculture International, 28, 2121-2137. .

López-Bueno, A., Mavian, C., Labella, A.M., Castro, D., Borrego, J.J., Alcami, A., & Alejo, A. (2016). Concurrence of Iridovirus, Polyomavirus, and a Unique Member of a New Group of Fish Papillomaviruses in Lymphocystis Disease-Affected Gilthead Sea Bream. Journal of Virology, 90 (19), 8768-8779.

Madduppa, H.H., Juterzenka, K.V., Syakir, M., & Kochzius, M. (2014). Socio-Economy of Marine Ornamental Fishery and Its Impact on The Population Structure of the Clown Anemone Fish Amphirion ocellaris and Its Anemones in Spermonde Archipelago, Indonesia. Ocean & Coastal Management, 100, 41-50.

Maganha, S.R.D.L., Cardoso, P.H.M., Balian, S.D.C., Queiroz, S.R.D.A., Fernandes, A.M., & Sousa, R.L.M.D. (2020). Detection and Molecular Characterization of Lymphocystivirus in Brazilian Ornamental Fish. Brazillian Journal of Microbiology, 51 (2), 531-535.

Palmer, L.J., Hogan, N.S., & Van den Heuvel, M.R. (2012). Phylogenetic Analysis and Molecular Methods for The Detection of Lymphocystis Disease Virus From Yellow Perch, Perca flavescens (Mitchell). Journal of Fish Diseases, 35 (9), 661-70.

Sihananto, B.S., Novita, H., Wianty, C., & Lusiastuti, A.M. (2019). A Case Study of Lymphocystis Virus Disease in Farmed Giant Snakehead (Chana Striata) in Mandiangin, South Kalimantan. Indonesian Aquaculture Journal, 14 (2), 75-81.

Tidona, C.A., & Darai, G. (1997). The Complete DNA Sequence of Lymphocystis Disease Virus. Virology, 230 (2), 207-216.

Walker, P.J., Siddell, S.G., Lefkowitz, E.J., Mushegian,

A.R., Adriaenssens, E.M., Alfenas-Zerbini, P.,

Davison, A.J., ....., & Zerbini, F.M. (2021). Changes to Virus Taxonomy and to The International Code of Virus Classification and Nomenclature Ratified by The International Committee on Taxonomy of Viruses. Archives of Virology, 166 (9), 2633-3648.

Xu, L., Feng, J., & Huang, Y. (2014). Identification of lymphocystis disease virus from paradise fish Macropodus opercularis (LCDV-PF). Archives of Virology,, 159 (9), 2445-9.

Zhang, Q.Y., Xiao, F., Xie, J., Li, Z.Q., & Gui, J.F. (2004). Complete Genome Sequence of Lymphocystis Disease Virus Isolated from China. Journal of Virology, 78 (13), 6982 – 6994.


Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

Creative Commons License
Indonesian Aquaculture Journal is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

View My Stats
p-ISSN: 0215-0883
e-ISSN: 2502-6577


Hasil gambar untuk isjd