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In-Vivo Test of Spirulina sp as Inducer of β-Actin In Cantang Grouper (Epinephelus fuscoguttatus-lanceolatus) Infected by Viral Nervous Necrosis

JFMR-Journal of Fisheries and Marine Research Vol 2, No 3 (2018): JFMR VOL 2 NO 3
Publisher : JFMR-Journal of Fisheries and Marine Research

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Abstract

Cantang grouper fish (Epinephelus fuscoguttatus-lanceolatus) is a result of hybridized fish between a female of tiger grouper fish (Epinephelus fuscoguttatus) and male of kertang grouper (Ephinephelus lanceolatus). In the development of cultivation, there are many problems, one of them infected with the class of virus Nodaviridae, namely Viral Nervous Necrosis (VNN). Fish had a defense against cellular immunity against the VNN virus by β-actin. The aims of this research to explore the crude extract of Spirulina sp as a β-actin inducer for the anti-inflammatory immune system in grouper fish against VNN attack. The method used in this research is experiment methods.  Crude extracts of Spirulina sp (33 μg/ml) were conducted by feeding orally to groupers, and VNN infections were conducted by feeding the already positive VNN meat. Detection of VNN using RT-PCR, however β-actin detection using PCR, and IHC in the organ of Cantang grouper fish. The results showed that the percentage of DAB value of control fish (14.0%), fish treated with Spirulina sp (25.7%), fish treated with VNN (31.9%), and fish treated with Spirulina sp extract and VNN infection (32.4%). The percentage of DAB values were indicated by the detection of the target gene β-actin. Immunity in fish increases with the addition of Spirulina sp. The increased β-actin expression may also be used as an indicator of a grouper's body defense against VNN infection.

Analysis of Genotype and Phylogenetic of Koi Herpesvirus (KHV) on Carp (Cyprinus carpio) in Lombok Island of West Nusa Tenggara Province Based on Thymidine Kinase Sequence

The Journal of Experimental Life Science Vol 8, No 1 (2018)
Publisher : Graduate School, University of Brawijaya

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Abstract

Koi herpesvirus (KHV) is a virus that usually infects carp and koi causing mass mortality, as well as economic and social loss. The results of varied test are often found in its development. Allegedly, there are mutation or genetic variation of KHV in Indonesia. Based on these problems, this research aims to understand the genetic variation and kindship of KHV isolate in the carp (Cyprinus carpio) in Lombok Island of West Nusa Tenggara Province based on thymidine kinase sequence. Water quality of fish pond was checked. KHV clinical symptom was checked visually in each fish sample, then sacrifices the organ (gill) for PCR analysis and DNA sequencing. The research results show that clinical symptoms that appear are pale fish gill and bleeding on the surface of the body. The temperature in the research site shows an optimum range for KHV development. The results electrophoresis of carp suspected with KHV shows five positives out of a total six samples. Genetically, KHV isolates from Lombok Island of West Nusa Tenggara Province is identical with the GenBank isolate code KHV-J/U/I and Indo_0C05WNT2.Keywords: Carp, Koi herpesvirus, Phylogenetic, Thymidine kinase.

THE EFFECT OF CRUDE PROTEIN HALIMEDA SP. ON CYPRINUS CARPIO INFECTED KOI HERPES VIRUS ON EXPRESSION OF MAJOR HISTOCOMPATIBILITY COMPLEX CLASS-1

JFMR-Journal of Fisheries and Marine Research Vol 1, No 1 (2017): JFMR
Publisher : JFMR-Journal of Fisheries and Marine Research

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Abstract

Halimeda sp. is a kind species of macroalgae that abundant grow in Indonesia. The utilization of Halimeda especially protein for disease prevention caused by the virus has not been done to the fish. The purpose of the study is to know the  treatment of crude protein Halimeda sp on  Cyprinus carpio infected by Koi Harvest Virus (KHV) on the expression of Major Histocompatibility Complex (MHC) class 1. MHC class-1 is one of the immune adaptive response on fish like this C. carpio. The methods are isolation of crude protein Halimeda sp,   purification  of  crude protein, Haemagglutination (HA) test and dot blot test. The result shows that the crude protein of Halimeda sp. can produce of adaptive immune response like MHC class 1. MHC class-1 has the function of  the immune system for maintenance of virus attacks directly. Responsible of treatment crude protein Halimeda sp. on C. carpio can be showed quantitatively using the dot blot test result. The  conclusion  is the crude protein of Halimeda sp. able to activate the MHC class I and to inhibit the proliferation of the KHV and maintenance of the fish cell of C. carpio..

The Characterization of Haemaglutinin Protein of Vili Vibrio alginolyticus as Adhesion Molecule on Epithelial Cells Receptor of Humpback Grouper (Cromileptes altivelis)

Jurnal Kedokteran Hewan Vol 2, No 1 (2008): J. Ked. Hewan
Publisher : Syiah Kuala University

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Abstract

Vibriosis was a crucial problem and a primer infection causing mortality of humpback grouper.Virulence factor of vibrio, such as vili, is an adhesion molecule protein based on haemaglutination test toblood serum of humpback grouper. This research was aimed to characterize an adhesion molecule of viliprotein of V. alginolyticus on epithelial cells of Humpback grouper as haemaglutinin protein. Themethods are laboratory exploration by isolate epithelial cells of Humpback grouper, characterize a receptorand test vili protein to adhesion molecule that is epithelial cell of Humpback grouper. The result of proteinhaemaglutinin test to vili V. alginolyticus show that an adhesion molecule with a molecule weight of38.98 kDa is an adhesin vili V. alginolyticus and a haemaglutinin. The adhesion pattern of V.alginolyticus on epithelial cells of Humpback grouper exposed to protein haemaglutinin of vili show bothdiffuse and localize pattern. Research conclusion was the haemaglutinin protein of vili V. alginolyticuswas an adhesin having a role to both virulence process of bacteria and attachment process to receptor ofepithelial cells of Humpback grouper.Keywords: adhesion molecule, epithelial cell, vili, V. alginolyticus, C. altivelis

The Analysis of Hepatopancreas Histologycal Damage in Neocallichirus karumba (Poore and Griffin) Shrimp Caused by Heavy Metal Pb Exposure in Madura Strait

The Journal of Experimental Life Science Vol 6, No 1 (2016)
Publisher : Graduate School, University of Brawijaya

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Abstract

Madura strait known as the second busiest shipping lanes in Indonesia. Human activities on the environment can influence the marine ecosystem derived from the household, fishery processing and capture fisheries. It can pollute coastal environments, one of which is heavy metal exposure such as Cd, Hg, Ca, As and Pb. These metals are greatly impacting the life of sea biota. The objective of this research, therefore, is to examine the hepatopancreas damage rate of shrimp Neocallichirus karumba due to the exposure of heavy metal Pb in Madura Strait. Method of research is by analyzing Pb content in water, sediment and shrimp by taking sample from 3 different stations. Hepatopancreas damage is then analyzed with SEM-EDX. Result of research indicates that at Station A, Pb pollution in water is the biggest and counted for 0.25 ppm, while those in sediment and shrimp are 5.85 ppm and 1.24 ppm. At Station B, Pb pollution in water is 0.19 ppm, whereas that in sediment is 5.51 ppm, but that in shrimp is 1.04 ppm. At Station C, Pb pollution in water counts for 0.18 ppm, and in sediment, it stands for 5.5 ppm but 0.02 ppm for shrimp. Result of analysis against hepatopancreas damage is explained as follows. At Station A, vacuolization is 20 % and Pb content in organ is 0.520 ppm. At Station B, the parameters are 10% and 0.196ppm. At Station C, it includes 15% and 0.173ppm. Organ damage is straightforwardly related to Pb content in water and sediment. Shrimp age is quite influential to the percentage of organ damage.Keywords: Heavy metal Pb pollution, hepatopancreas organ damage, Madura Strait, N. karumba shrimp.

Identification of Plankton on Fish Pond of Oreochromis niloticus Infected by Viral Nervous Necrosis

Research Journal of Life Science Vol 3, No 2 (2016)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat, Universitas Brawijaya

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Abstract

The purpose of study is to determine the type of plankton found in cultivation pond of nile tilapia infected by VNN. The method is to identify the plankton in the water pond and testing the VNN that infects nile tilapia related to plankton eaten using Polymerase Chain Reaction (PCR). VNN identification results on the nile tilapia digestive tract stomach and intestines as well as plankton found in the stomach and intestines indicate VNN positively. Plankton found in the digestive tract of fish is division of Chlorophyta, Bacillariophyta, phylum Rotifera and Crustacea. It indicates that nile tilapia infected by VNN both on the gastrointestinal tract organs, stomatch or intestines and plankton were found with positive VNN digestive duct, while plankton found in water pond is VNN negative. The VNN infection depends on the nature of specifications VNN on nile tilapia and the plankton in the nile tilapia digestive duct.

Assessing the Genotoxic Potentials of Methomyl-based Pesticide in Tilapia (Oreochromis niloticus) Using Micronucleus Assay

The Journal of Experimental Life Science Vol 7, No 2 (2017)
Publisher : Graduate School, University of Brawijaya

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Abstract

Pesticides are recognized as serious pollutants in the aquatic environment with the potential to cause genotoxic on the aquatic organism, especially fish. The micronucleus (MN) assay has been used to evaluate genotoxicity of many compounds in polluted ecosystems such pesticides. The aim of this study to determine genotoxic effect of methomyl-based pesticide on tilapia (Oreochromis niloticus). Fish were exposed to six different concentrations base on range finding test (0 ppm, 3.2 ppm, 4.2 ppm, 6.5 ppm, 8.7 ppm and 10 ppm) of methomyl-based pesticide. The micronucleus were collected from peripheral blood erythrocyte of fish after 96 h exposure. Peripheral blood samples smears were stained with Giemsa, MN frequencies were counted and statistically analyzed using one-way ANOVA. The result of this study showed after 96 hours exposed to methomyl-based pesticide, at concentration 0 ppm causes 0% mortality, at concentration to 3.2 ppm causes 30% mortality, at concentration 4.2 ppm causes 60% mortality, at concentration 6.5 ppm causes 70% mortality, at concentration 8.7 ppm causes 80% mortality, at concentration 10 ppm causes 100% mortality of fish test. Lethal Concentration 50 (LC50 - 96 hours) of methomy-base pesticide towards tilapia (O. niloticus) is 4.015 ppm. Through micronuclei assay during 96 hour exposure of methomyl-based pesticide, the result shows that frequencies of micronuclei in erythrocyte of fish test at concentration at 0ppm is 12‰, 18‰ and 16‰; at concentration at 3.2ppm is 33‰, 26‰ and 29‰; at concentration at 4.2ppm is 41‰, 38‰ and 46‰; at concentration at 6.5ppm is 68‰, 81‰ and 82‰; at concentration 8.7ppm is 133‰, 130‰ and 137‰; at concentration 10ppm is 163‰, 166‰ and 156‰. It revealed that methomyl-based pesticide exposure induced after 96 h significantly (P<0.05) increased genotoxic potentials simultaneous with increased concentration.Keywords: Genotoxic, Methomyl, Micronucleus Assay, Pesticide, Tilapia.

Bobot Molekul Resptor Salmonella Typhosa pada Kultur Sel Endotel Vena Umbilicalis Manusia

Jurnal Kedokteran Brawijaya Vol 18, No 1 (2002)
Publisher : Fakultas Kedokteran Universitas Brawijaya

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Abstract

Penyakit demam tifoid yang disebabkan oleh bakteri Salmonella (S) Typhosa masih merupakan problema kesehatan terutama di negara yang sedang berkembang termasuk Indonesia. Pada patogenesis terjadinya bakteriemi dan mengenai pelekatan bakteri tersebut terhadap sel endotel masih belum ada kejelasan. Tujuan penelitian ini untuk mengetahui kemampuan pelekatan S.typhosa dan bobot molekul reseptornya pada sel endotel. Penelitian ini dikerjakan dengan memakai Human Umbilical Vein Endothelial Culture (HUVEC), sedangkan sebagai model sel pelekat bakteri yang digunakan adalah model indeks adhesi. Untuk mengetahui indeks adhesi dilakukan pemaparan S.typhosa terhadap HUVEC. Matriks ekstraseluler  diekstrasi dengan cara solubiliasi HUVEC dengan larutan N-Octyl-β-D Glucopyranoside (NOG) 0,05%. Penentuan bobot molekul matriks sektraseluler yang dikerjakan dengan SDS-PAGE ternyata memiliki bobot 66kDa. Pemurnian protein matriks ekstrak seluler dipakai cara elektroelusi. Ketentuan bobot molekul reseptor ditentukan dengan evaluasi korelasi anatara indeks adhesi bakteri dengan dosis protein molekul reseptor yag diencerkan setengahnya secara seri yang dilekatkan pada HUVEC. Kesimpulan dari evaluasi tersebut dengan menggunakan tes korelasi menunjukkan bobot molekul matriks ekstrak seluler 66kDa HUVEC merupakan molekul reseptornya S.typhosa M223 (p<0,05).

Expression of Hsp70 and β-actin Genes as The Immune Response against Viral Nervous Necrosis that Infected Asian Seabass (Lates calcalifer)

The Journal of Experimental Life Science Vol 8, No 3 (2018)
Publisher : Graduate School, University of Brawijaya

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Abstract

The viral nervous necrosis (VNN) is betanodavirus of the family Nodaviridae with acute infection and associated with high levels mortality up to 100% of numerous larval-stage marine and freshwater fish species. The danger signals of viral infection that are capable of activating APCs (Antigen Presenting Cells), furthermore produced molecules protein receptors such as interferon and heat shock proteins. Stimulating interferon type I (IFN I) induce several antiviral molecules, further binding the actin cytoskeleton to reach the site of infection. This study demonstrated a correlation between increased levels of Hsp70 (heat shock proteins) and actin filamentous (β-actin) within invasion wild-type isolate of ssRNA VNN from Asian Seabass (L. calcalifer) juvenile. Furthermore, expression of Hsp70 and actin as an indicator or biomarker of stressed states in fish. The reverse transcriptase polymerase chain reaction (RT-PCR) method used to finding expression Hsp70 and β-actin. Whereas nested RT-PCR used for VNN genes.  The up-regulation of Hsp70 was observed on the brain tissue higher than eye tissue of sample positive infected VNN. Whereas different expression of β-actin, β-actin receptor expression tends to be stable in the organs of the brain and eye, both invaded VNN or normal, while on the eye, the ratio slightly increased based on the intensity of the band.  The research shows that altered expression of heat shock protein 70 and beta-actin gene receptors in the target organs that response to invasion VNN at Asian seabass (L. calcalifer) juvenile. Alteration expression of heat shock protein 70 and beta-actin can be used as an indicator or biomarker of stress cells, especially from VNN invasion at fish.Keywords: Asian seabass, β-actin, Hsp70, Immune response, Viral nervous necrosis.

ICTYOFAUNA IN THE DOWNSTREAM OF KLAWING RIVER, PURBALINGGA DISTRICT CENTRAL JAVA

Samakia: Jurnal Ilmu Perikanan Vol 9 No 2 (2018): Samakia: Jurnal Ilmu Perikanan
Publisher : Faculty of Science and Technology University Ibrahimy

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Abstract

The status and potential of ichtyofauna resources in public waters, especially downstream of the river, must be monitored and evaluated frequently. Land use along the downstream river is often used as mining and industrial activities. The purpose of this study is to find out fish resources in the lower reaches of the Klawing Purbalingga River in Central Java. The study was conducted in January-March 2017 with descriptive exploratory method. Sampling using gill nets with the help of local fishermen. The fish obtained were identified and grouped by family to the species level. The results showed that in the lower reaches of the Klawing River there were species diversity consisting of 13 species, 7 families and 13 genera. Ichtyofauna from Cyprinidae family dominate as many as 6 species followed by Bagridae family 2 species. One introduced species of Osphronemus gouramy and one invasive alien fish species Hypostomus plecostomus and other species were indigenous species.