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Ketut Mahardika

Research Institute for Mariculture, Gondol, Bali

Author-ID : 3672502

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology

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QUANTITATIVE HISTOPATHOLOGICAL ANALYSIS OF ENLARGED CELLS DERIVED FROM HUMPBACK GROUPER, Cromileptes altivelis INFECTED WITH GROUPER SLEEPY DISEASE IRIDOVIRUS (GSDIV) Indah Mastuti; Yasmina Nirmala Asih; Ketut Mahardika
Indonesian Aquaculture Journal Vol 5, No 2 (2010): (December 2010)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Pathognomonic sign of grouper sleepy disease iridovirus (GSDIV) was proposed to be the formation of enlarged cells and necrotic cells, in which under electron microscope, it is revealed to be the inclusion body bearing cells (IBCs) and necrotic cells containing GSDIV viral particles. Spleen and kidney tissues are the major sites for formation of enlarged cells. This paper described the result of histopatological analysis of enlarged cells found in the spleen and kidney of moribund fish after GSDIV challenge. A pathogenicity test was conducted on fish stocked in two tanks for infected groups and the other two tanks for uninfected control groups (15 fish per tank). The infected groups were injected intramuscularly with 0.1 ml of the viral inoculum. The uninfected groups were injected with the same amount of EMEM-2. The GSDIV-infected humpback grouper began to die after 6 days post infection and all died after 7 dpi, excluding one fish which had survived until the end of experimental infection periods (93% to 100% mortality). All of the diseased fish showed massive formation of enlarged cells in their spleen, head kidney and trunk kidney. The largest number of enlarged cells was observed on head kidneys and subsequently followed by spleens, trunk kidney (2.0-200.3/field of view). This result suggested that the number of enlarged cells in the affected organs was not the direct factor that led to the mortality of fish.

ULTRASTRUCTURAL ANALYSIS OF GRUNT FIN (GF) CELLS TREATED WITH RED SEA BREAM IRIDOVIRUS (RSIV; family Iridoviridae, genus Megalocytivirus) IN COMBINATIONS WITH INTERFERONS AND SPLENIC SUBSTANCES Ketut Mahardika; Teruo Miyazaki
Indonesian Aquaculture Journal Vol 5, No 1 (2010): (June 2010)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

The genus Megalocytivirus in the family of Iridoviridae encompasses isolate of red sea bream iridovirus (RSIV). In the present study, grunt fin (GF) cells were treated with red sea bream iridovirus (RSIV) in combinations with interferons (IFNs) and splenic substances. The viral titer in the combination with primary splenic substance was higher than the other combinations of 10-1 and 10-2 diluted splenic substances, and the positive control. The viral titer was not decreased by all combinations with recombinant murine interferon-α (rMuIFN-α), recombinant murine IFN-β (rMuIFN-β), and recombinant feline interferon-ω (rFeIFN-ω). Electron microscopy revealed inclusion body bearing cells (IBCs) and enlarged cells allowing virus propagation within the intracytoplasmic virus assembly site (VAS). Most were enlarged cells. These enlarged cells were divided into three cell types. Cells of Type II, which contained many mature virions within the VAS, were numerous in number in all treated cells. Cells of Type I allowing assembly of few virions and cells of Type III containing many immature viral particles were rather fewer in number. Their percentage was almost the same in all combinations with the splenic substances and IFNs. These results determined in in vitro treatment with IFNs did not prevent viral replication of RSIV, as well as the splenic substances which were derived from the RSIV-infected spleen of red sea bream did not contain any factors to disturb RSIV replication.

ULTRASTRUCTURAL FEATURE OF KOI HERPESVIRUS (KHV) INFECTED CULTURED KOI FIN (KF-1) CELLS Ketut Mahardika; Masahiro Yasuda
Indonesian Aquaculture Journal Vol 6, No 2 (2011): (December 2011)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Koi herpesvirus (KHV), may cause significant morbidity and mortality in common carp (Cyprinus carpio). In the present study, an electron microscopic (EM) was performed on KHV-infected cultured koi fin (KF-1) to document the ultrastructure of the lesions. Viral particles were firstly evident in the nucleus. These viral particles observed as immature capsids and nucleocapsids. Many non-enveloped nucleocapsids have moved from the nucleus into the cell cytoplasm. The formation of subviral particles and virions, which comprised, in turn, an electron dense core, capsids with a hexagonal outline, the tegument was evident in the cytoplasm. And then, the virions with the enveloped tegument budded through the intracytoplasmic membrane. Based on EM results, the definitive pathological change was similar as those in the Family Herpesviridae.

PATHOGENECITY OF GROUPER SLEEPY DISEASE IRIDOVIRUS (GSDIV: Megalocytivirus, FAMILY Iridoviridae) TO CORAL TROUT GROUPER Plectrophomus leopardus Ketut Mahardika; Ahmad Muzaki; Ketut Suwirya
Indonesian Aquaculture Journal Vol 4, No 2 (2009): (December 2009)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Grouper sleepy disease iridovirus (GSDIV), a member of the genus Megalocytivirus in the family Iridoviridae, has been known to cause large scale mortalities resulting in severe economic losses in grouper industries in south-east Asia including Indonesia. In this study, experimental infection of coral trout grouper Plectrophomus indicus with GSDIV was performed to evaluate the viral pathogenecity to this fish species. After virus exposure, the mortalities of coral trout grouper injected with primary and 10-1 dilution of spleen homogenates derived from tiger grouper Epinephelus fuscoguttatus were 100% and 90%, respectively. Histopathology revealed that moribund fish receiving GSDIV inoculum displayed massive formation of enlarged cells in the spleen and hematopoitic tissues. Under electron microscopy, the enlarged cells were observed as inclusion body bearing cells (IBCs) and necrotic cells allowing virus propagation within an intracytoplasmic virus assembly site (VAS). GSDIV virions were 167-200 nm in size. These findings confirmed that GSDIV has severe pathogenicity to coral trout grouper and IBCs as well as necrotic cells were determined to be the pathognomonic sign of megalocytivirus-infected coral trout grouper.

ELECTRON MICROSCOPIC STUDY ON ENLARGED CELLS OF RED SEA BREAM, Pagrus major INFECTED BY THE RED SEA BREAM IRIDOVIRUS (RSIV, GENUS Megalocytivirus, FAMILY Iridoviridae) Ketut Mahardika
Indonesian Aquaculture Journal Vol 4, No 1 (2009): (June 2009)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Most histopathologycal studies of the red sea bream iridovirus (RSIV) disease in red sea bream have been performed by studying enlarged cells as well as necrotized cells in the spleen and other organs. These enlarged cells have been named as inclusion body bearing cells (IBCs). However, few information is available about detail of ultrastructural features of IBCs produced in the target organs of RSIV-infected fish. In the present study, details of ultrastructural features of IBCs that were produced in the spleen tissue of naturally RSIV-infected red sea bream were investigated under electron microscope. Under electron microscope, RSIV-infected red sea bream had the presence of two types of IBCs: typical IBCs allowing virus assembly within viral assembly site (VAS), and atypical IBCs which degenerate organelles without virus assembly. Other infected-cells were observed as necrotized cells forming intracytoplasmic VAS with large numbers of virions, but without the formation of the distinct inclusion body. Morphogenesis steps on RSIV-infected red sea bream were observed as filamentous-filed virions, partially-filled virions and complete virions with 145-150 nm in size. These findings confirmed that RSIV-infected red sea bream were characterized by formation of typical and atypical IBCs as well as necrotized cells.

Co-Authors Ahmad Muzaki Indah Mastuti Ketut Suwirya Masahiro Yasuda Teruo Miyazaki Yasmina Nirmala Asih

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