Vaccines Application in Indonesian Aquaculture

I. Background Fish is the largest creature in the waters and biodiversity which a high-protein food. Fresh fish contains 16-24% protein, 0.2 to 2.2% fat, vitamins, minerals and carbohydrates (Suseno, 2004). Aquaculture has developed because it has a good chance in the domestic and international market, such as the European Union, ASEAN, Hong Kong, China and Japan. One of the main obstacles aquaculture development, both in freshwater, brackish or sea ecosystems is a crop failure due to infectious and non-infectious disease. The problems of these diseases are very important because they can lead to decreased production, quality of fish and even death (Kabata, 1985). In freshwater commodity, fish farming constraints due to Aeromonas infection began to arise since 1980 with a loss of about US$ 2 million. This is a pathogenic bacterium, spread very rapidly at high stocking density and can lead to death until reaches 90% of population (Kabata, 1985). Another obstacle to the cultivation of freshwater is the presence of herpes disease in carp and koi. Koi Herpes Virus (KHV) is a new emerging disease known to cause gill and skin damage in koi and carp (Cyprinus carpio). The disease suspected to have been introduced into Indonesia through importation of koi from Hongkong. It is currently occurring in Indonesia since March 2002 starting in the area of Blitar in East Java. Since then it has been spreading rapidly throughout Java Island, Bali, southern part of Sumatra, East Kalimantan, and Central Sulawesi (Sunarto et al., 2005). The history of KHV in Indonesia including the first 3-episodes of the outbreak was chronologically described by Sunarto et al. (2002). The first episode of mass mortalities of cultured koi (Cyprinus carpio) was recorded in March 2002 in Blitar, East Java. It occurred after heavy rains among new fishes introduced from Surabaya, the capital city of East Java. The fish were imported from China through Hongkong in December 2001 and January 2002. Blitar is well known as the centre for koi production in the country. The koi including the infected one were distributed over the country, with to Central Java, West Java and Jakarta as the main market (Sunarto et al., 2005). The second disease outbreak occurred in cultured common carp (Cyprinus carpio) during the end of April 2002 in Subang regency, West Java. The gross signs of the diseased common carp were extremely similar with that observed previously in koi carp. Subang is one of the production centers of common carp in West Java. Since then the outbreaks spread to neighboring regencies mainly through fish movements (Sunarto et al., 2005). The third episode of the outbreak occurred in end of May to early of June 2002 in cultured common carp in floating net cage at Cirata Reservoir, West Java. Weeks before the outbreak, farmers introduced common carp from Subang region due to the low price of fish (Sunarto et al., 2005).

The fourth episode of the outbreak occurred in February 2003. The outbreak affected cultured common carp in Lubuk Lingau regency, South Sumatera. The gross signs of the diseased common carp were extremely similar with that observed previously in koi and carp in Java islands. Common carp farms at Lubuk Lingau were infected with the disease coming from Cirata Reservoir, West Java through fish transfer by traders. The outbreak then spread to neighboring district and province including Bengkulu in the south and Jambi in the west (Sunarto et al., 2005). The first report regarding the economic losses due to the outbreak was made by the head of the Association of Ornamental Fish Culture of Blitar regency, East Java. They reported that in Blitar alone, the outbreak destroyed high quality koi carp belong to 5,000 fish farmers with economic losses more than IDR 5 billions (US$ 0.5 millions) within the first 3 months period of the outbreak. As of July 2002, the NACA¶s Task Force estimated that the loss revenue of the sector and the socio-economic impact to the rural farming communities was in the region of US$ 5 millions. As the outbreaks continued and spreading to new areas, the socioeconomic impact due to the outbreaks was escalated. Directorate of Fish Health and Environment (DFHE) estimated that as of December 2002 and 2003, losses due to KHV were US$10 million and US$15 million, respectively (Sunarto et al., 2005). Central Freshwater Aquaculture Development (Balai Besar Perikanan Budidaya Air Tawar, BBPBAT) monitoring results (2002-2005), showed that KHV virus attack all stages of common carps or koi. Based on data from the spread of KHV in Indonesia, almost all regions, there are strains of carp attack by KHV. However, there are not yet data about the percentage difference in mortality between strains of carp. The mortality percentage differences between strains of carps may be related to KHV tolerance (Mudjiutami, 2007). Results of monitoring and surveillance Directorate of Pests Fish Diseases, Directorate General of Aquaculture Ministry of Marine Fishery until 2009 showed the expansion of the spread of KHV in Indonesia that has got to Sumatra (kab. Tanah Datar, Lima Puluh Kota and Sawah Lunto), Jambi (Kab. Kerinci), NTB (West Lombok), South Kalimantan, East and West Kalimantan. In brackish and marine water commodities, a major cause disease that affects fish farming and wild fish are Vibrio sp. This disease has been recognized since 1718 in Italy, with many epizootics documented throughout the 19th century. Types of bacterial disease found in fish farming include caudal fin ulcer and red mouth disease. The bacterial isolation and identification found some types of bacteria are closely associated with cases of suspected disease, namely Vibrio alginolyticus, V. algosus, V. anguillarum and V. fuscus. Vibriosis can be either acute or chronic depending on the type of Vibrio that attack, host condition, and environmental conditions. In general, this disease can cause death up to 100%. In addition there is an also viral disease that causes the collapse of marine fish and shrimp cultivation, namely, Viral Nervous Necrosis (VNN) and White Spot Syndrome Virus (WSSV). VNN has spread in Japan, Korea, China, Southeast Asia, northern Australia, Austria, Iran, Israel, Greece, France, Norway, Canada and America. These diseases are extremely malignant, so rapidly spreading in various parts of earth. Thus, these pathogens were greatly feared by all countries that have fishing industry. These pathogens can cause mass mortality of farmed fish in a relatively short time (Chi, 2006 in Bimantara, 2009). Most
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II. antibiotic free.) in Southeast Asia. In an effort to control fish disease. the National Residue Control Plan program was launched on 2010 in order to support quality assurance and food safety in fish cultivation to obtain residue -free fishery products.1 Aeromonas hydrophila A. In some countries. because only by providing one-two times were able to increase the resilience of fish's body until the end of maintenance period. the use of vibriosis vaccines which added with immunostimulant showed survival rate reached 100%. still using antibiotics as a curative action. while Aeromonas hydrophila vaccines showed 80-90% with 6-12 months protection. VNN be a major challenge in aquaculture industry because of the high levels frequency of disease occurrence that can lead mortality approaching 100% and wide distribution both in warm and cold waters as media of various kinds of fish live. product quality. chemical. This pathogen attack fishes which reared in floating net cage and fish larvae are still kept in hatchery which spread across Indonesia. However. the government issued a regulation of Minister of Marine Affairs and Fisheries Republic of Indonesia Number 02/men/2007 about monitoring of drug. the vaccines use can increase fish production to reach 45. The European Union has some regulations related to residue limits of pharmacologically active substances in foodstuffs of animal origin. like USA. Asian seabass (Lates calcarifer Bloch. and many others. Epinephelus spp. Kamiso on 2007. These techniques are environmentally friendly and more efficient. Japan and Europe has produced several commercial vaccines.) in Northern Australia.5%. Seabass (Dicentrarchus labrax L. The realizations of this program are fish drug and chemical controlled with feed antibiotics free and monitoring residues implemented in fish cultivation. and environmentally friendly. An alternative disease control in aquaculture can be done with health care management by improving fish immune through vaccination and immunostimulation. Under those regulations. hydrophila vaccine preparation needed to consider various aspects. Vibrio vaccine use in Salmon. compared with non-vaccinated groupers are only 12. this is no longer permitted as the policy of "say no to antibiotics" and the demands of global markets of food safety. namely: Grouper. such as heterogeneous antigens. one of the most recent regulation is EC 470-2009. relatively low immunity. To follow up this regulation. Recombinant protein vaccine use in Japan showed VNN infection resistance with 10-65% mortality rates and showed a high antibody titer up to 100 days. Striped jack (Pseudocaranx dentex) in Japan. Based on Willoughby data on 1999. India and Australia. Canada. biological. in Southeast Asia and Japan. Some research of the vaccines used that have been done and showing good results. Vaccine Research in Indonesia 2. In Indonesia. Red Drum (Sciaenops ocellatus) in Korea. Based on research conducted by Prof. Golden Grey mullet (Liza auratus) in Iran.of fishes being attacked by VNN in Asia. There are two types of vaccine Page 3 of 21 .000 tons on 1997 in Norway. and contaminants residues in fish cultivation. showing 100% Relative percent survival (RPS). and its application in field.

hydrophila vaccine has been pioneered by The Bogor Research Institute for Freshwater Aquaculture (BRPBAT) since 1983. The results showed that A.. several tests carried out on A. 2002). The vaccine contents about 87% of fish. 2000. (2004) has A. Research on A. membrane proteins have a complex role in many processes and cell physiology. about the potential immunogenic and virulence in vitro and in vivo to get one antigens candidate that have cross-react potential to other isolates. which can easily be recognized as an element of foreign substances in host defense system. In the early 1980s an epidemic of bacterial meningitis in a freshwater fish that attacks various species of fish in various sizes. Plague originated from West Java area and eventually spread to all parts of Indonesia with loss rate is estimated to reach hundreds of million rupiah. cheap. Since then the research and the study of biology. Increasing of an immune response does not mean that immunity has been achieved 2. Vaccination for fish diseases control has been developed since the last two decades. 1992). 2. (1984) has managed to increase Japanese eel (Anguilla japonica) immune with A. Factors to be considered in developing an effective vaccine are: 1. 1999). ordalii polyvalent vaccines for salmon and trout. siderophore and adhesin. This is because. Results showed that POM has proven to be a component of a potential vaccine for fish diseases control. Stimulation of memory cells is important to establish protection in future and this depends on the pathogen incubation period. hydrophila vaccine test intramuscularly with various boosters. Page 4 of 21 . hydrophila monovalent vaccine. hydrophila. but still a few of vaccines for fish diseases that are commercially available. hydrophila. The pathology team pioneered and exploits fish immunology and uses it as a major breakthrough in prevention of fish diseases. namely live vaccines or attenuated bacterial vaccine and death cells or only certain parts of attenuated (Ward. Outer membrane layer is basically composed of protein. efficient and effective. Results of epidemiological studies indicate that one type of pathogen that considered responsible for these cases is A. character and mechanism of the disease are done intensively to obtain a rational prevention technology. and antibody titer. 1982). The discovery of prevention techniques such as vaccines begins from the discovery of some evidence that in fish¶s body has immunoglobulin-like which have ability to induce immunity that can be stimulated either through artificial or natural. Qian et al. fat and sugar. outer membrane plays as an important role in infection and pathogenicity level to the hosts (Tsolis. Song et al. Among these components. Vibriosis Vaccine is an active ingredient that is used to stimulate active immunity in animals and humans (Kuby.. hydrophila isolates. according to Finlay & Falkow (1997) in Desrina et al. safe. hydrophila cell debris vaccine can increase level of relative protection. survival rate. The first commercial vibriosis vaccines are V.2. Based on this evidence. Mulia et al. In gram-negative pathogenic bacteria.A. which is very attractive as a candidate used to develop vaccines and diagnostic kits (Pizza et al. anguillarum and V. Protein outer membranes (POM) which have been identified include porin.

shows the results that administration of anti-KHV DNA vaccine with 12.5 to 63. research and vaccine development continues to create new vaccines.3%. DNA vaccines are a new generation of vaccines that obtained with encoding plasmid DNA that are immunogenic to the host. Prior to successfully invade its host. Based on the results of the Fish Drug Commission Meeting on January 13. Challenge test at common carp which had been given anti-KHV DNA vaccine on laboratory scale. but also the cellular immune response. KHV disease prevention using vaccines have been killed developed in BBPBAT Sukabumi on 2007 to 2009. coli were then used as a vaccine (Santika and Sri Nuryati. Mucus has the ability to agglutinate antigen chemically. the pathogen has to deal first with the physical and chemical body's defense barrier. 2003). After that the pathogen must be able to pass through the skin or scales prior to the scales fish. coli plasmid.5% / 100 l dose. was able to maintain the survival rate of common carp up to 72-96%. Plasmids were then transformed and propagation in some E. 2009). Fish and Environmental Health Directorate of the Directorate General of Aquaculture Ministry of Marine Affairs and Page 5 of 21 . 2. carried out by BBPBAT Sukabumi in cooperation with the Bogor Institute of Agriculture. 2009). KHV Recently. Fish have non-specific and specific healthy immune system. In addition a private company has issued a vaccine for the KHV disease. Pathogens must penetrate the mucus and the mucus barrier in the outer parts of the body. DNA vaccines can cause not only the humoral immune response. by giving 5 g of 73.3. POM vaccines Research has been conducted by Desrina (2006). DNA vaccine is a breakthrough experimental technique to protect the organism against disease by injecting pure DNA that contains no protein or peripheral material not expected to generate immunological responses. Unstable the immunogenic material gill phase as a source of virus is still a constraint that affects the success of conventional vaccination. 2010. After this section passes the pathogen has to deal with other non-specific defense system in the body (Tort et al.43 kDa of POM vaccine by intraperitoneal and able to protect the tiger grouper seed with 50% RPS.. coli bacteria. can be purified in large quantities and because it is stable at room temperature or under high temperature. does not require refrigeration.(2007) that many virulence factors and pathogenicity made its code so that POM in outer membrane is specific and strongly immunogenic. after 30 days of challenge test with active KHV (Santika and Sri Nuryati. Plasmid isolation products from cultures of E. Therefore a new research carried out using DNA vaccines. KV3 branded vaccines already passed the field test and ready to use. Test results of the vaccine in field and laboratory conditions (floating net cage) gives the results of increased survival rate ranged from 59. the first in Indonesia. has the ability to protect against infection. The vaccine is the result of genetic engineering in which the viral genes sequences that is immunogenic inserted into E.

2." Palar said. and manufacturing. Greedy also stated that EU law does not permit the use of this vaccine.Pi. it has a major business to develop aquaculture vaccines. According to Palar Batubara on 2010. the result is the survival level of harvested carp amounted to 95%. a field test conducted over two years in floating net cage at Cirata and Jatiluhur Reservoir. research. This vaccine is a live attenuated vaccine (attenuated vaccine). West Java.S. KoVax has a patent for KHV vaccines and diagnostic tools from the Hebrews University. Non-vaccinated fish before the challenge test are death. The companies from the United States (U. Results of reverse transcriptase PCR analysis showed that the gene contained by the vaccine expressed in the vaccinated fish which means that the vaccine is active. through laboratory-scale challenge test the vaccine can increase survival of common carp more than 95% for one month after vaccination. in marketing the KV3 vaccine for the Asian region including Indonesia. VNN Betanodaviruses (Nodaviridae family) are agents which cause viral nervous necrosis (VNN) or viral encephalopathy and retinopathy (VER) in various marine fish farming activities Page 6 of 21 . The letter states KV3 vaccine from a private company already graduated a field test. "Tests conducted during the second cycle of carp rearing. the KV3 vaccine is being sought in order to obtain SNI (Indonesian National Standard). however the price is still considered to be very expensive and from the testing that some indication of the virus is still virulent. For while the product is sold to farmers in the form of seeds that have been vaccinated. the company is the sole agent of KoVax Ltd. From the results expected if the farmer had seen and felt cultivation increased survival of carp seed that has been vaccinated it will generate confidence. DNA vaccine is an alternative effective and safe vaccine. Further explained.. Through research sponsored by the Department of Marine and Fisheries of West Java Province (year 2008) and Doctorate Grant research program by the SPS-IPB (year 2009). attenuated virus vaccines such as vaccines above has been available. 2010). In order to continue to strengthen the recognition of product quality KV3 vaccine. This type of Vaccines has a weakness that potential for infection if the attenuation is not perfect done (Nuryati. this step is done to reduce the risk of failure of the vaccination process. Israel.Fisheries issued a letter number 429/DPB/PB430DA/1/2010. This shows that DNA vaccines are very effective to cope with KHV virus attack. Sri Nuryati.4.Si which is a student of Veterinary Science Doctoral PS-SPS IPB. Within one month ahead was to be distributed to the cultivators of vaccinated carp seed size 3-5 cm per seed (age one month). which at the same time teaching staff at the Aquaculture Department of Fisheries and Marine Science Faculty of Bogor Agricultural University has made a DNA vaccine for KHV. S. Attenuated vaccine does have limitations that can be enhanced by a DNA vaccine. Therefore Greedy (2008) in Nuryati (2010) did not recommend the use of this KV3 vaccine in fish. M. Center for Freshwater Aquaculture Development Sukabumi on 2009 has been test the efficacy of commercial vaccines KV3 for koi and common carp. Furthermore. According to Alimuddin (2010).).

and these infections commonly result in high mortality (Yukio. Betanodaviruses is a small virus. In the development of larvae. both types are highly malignant in infected fish. yellow tang Zebrasoma flavesenes).). and two types freshwater fish (South American leaf fish and red piranhas Monocirrhus polyacanthus Pygocentrus nattereri). Red drum (Sciaenops ocellatus) mass mortality in hatchery associated with betanodavirus (Chi. China. VNN attack symptoms were first reported to attack grouper (Epinephelus septemfasciatus). no capsid with a genome consisting of double helix. blue ribbon eel Rhinomuraena quaesita. and BFNNV (barfin flounder nervous necrosis virus). This disease occurs mainly during the period of hatchery fish larvae and the cultivation process is under way (Dennis et al. Nodaviruses is icosahedral viruses that are not wrapped with a genome consist of two single helix RNAs13. 2006) VNN is a disease listed by Office International des Epizooties (OIE). Piscine nodavirus infection is a potential threat to cause damage to many species of fish aquaculture in the region. VNN attacks among population in marine fish farming can occur with the transmission vertically or horizontally. Identification of virus causing VNN is obtained Nodaviridae family members to investigate the nucleic acid and structural proteins of the Pseudocaranx dentex virus larvae. 2006). marked on the spine are points Page 7 of 21 . the first vacuolation need to be observed on spine.. three spot damsel Dascyllus trimaculatus. Viruses of this type are the agent that causes betanodavirus VNN. In subclinic. There are two types of Nodaviridae family. Thailand. In Korea. Singapore. fish and invertebrate in aquariums infection by inoculum betanodaviruses. Piscine nodaviruses (betanodaviruses) infected more than 30 species marine fish larvae. especially during juvennil. Milkfish Chanos chanos. look down on fish Selene vomer. and Indonesia. Monocentris japonica pinecone fish. RGNNV (redspotted grouper nervous necrosis virus). TPNNV (tiger puffer nervous necrosis virus). 2007). Betanodaviruses are causatif agent of VNN in marine fish farming. By phylogenetic analysis. Betanodaviruses (Nodarideae family) is the agent causing VNN in marine fish cultivation. namely Alphanodavirus and Betanodavirus. a devastating disease of marine fish farming industry worldwide (Chi. 2006. Recently been documented the outbreaks of VNN between hatchery of the orangespotted grouper and asian seabass in Philippines. Positive PCR test results obtained from the brain eight species of marine fish (shrimp fish Aeoliscus strigatus. spherical. Brain tissue of fish and other invertebrate animals have been tested with Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) with the aim to detect betanodavirus. swim bladder damage. isolation from orange-spotted grouper and asian seabass have RGNNV genotype (unpublished data). Japanese anchovy Engraulis japonicus. dorsal fin. Piscine nodavirus infections have been associated with high mortality rates in grouper cultivation in Taiwan. then brain condition and within retina. Piscine nodaviruses can be classified into four genotypes based on nucleotide sequence of coat protein genes: SJNNV (striped jack nervous necrosis virus). PCR detection level was 11/237 (4.around the world. which is marine fisheries production major problem in the world.64%). a marine invertebrate species (spiny lobster Pamulirus versicolor).

1998). transported through the network Axon spine on spine. Neurotropisme indications of VNN attack might gain access to CNS through peripheral nerves.. and family Nimaviridae. The third sequence is almost the same isolates. WSSV is described as non-occluded baculovirus.. Thie'ry et al. AF369029) and 307. Inouye et al.. 1996. et al. melanosis and diminished appetite. Prevention is more effective in controlling viral diseases. and potentially lethal for most commercially cultivated penaeid shrimp species (OIE 2003). There are no other types of antibiotics and chemotherapy which can be used for viral diseases treatment. 1997. Durand et al. VNN can attack CNS via blood circulation as starting point injection (Dennis et al. 2001. WSSV genome size has been reported for different isolates: 305. DNA vaccine. Via intramuscular (IM). Common VNN clinical sign in several fish species namely swimming behavior erratic.967 bp (GenBank Accession No. young fish larvae which attacked by viruses can be detected in skin epithelial cells and intestinal epithelium. To obtain the vaccine in suppressing malignancy VNN can be obtained from a variety of ways. with differences in size Page 8 of 21 . AF332093). 1997. 1996). VNN through peripheral nervous system in muscular tissue edge.to proliferation of VNN initial location (Nguyen et al. So we need a vaccine that can be used to boost immune system to be more resistant organisms from pathogenic viral infection attacks. including: recombinant vaccines. Death (mortality) achieved a cumulative 34% and 56% for 10 weeks. 2. which simultaneously with Central Nervous System (CNS) nerve cells as early stages of infection or inflammation by VNN. AF440570) for viruses isolated from China. 1994.. For the procurement of vaccine virus is very helpful in suppressing the impact caused by the virus that causes a variety of viral diseases. 2006). Kanchanaphum et al. elliptical rods. WSSV WSSV has a broad host range among decapod crustaceans (Lo et al. and float to stomach caused by swim bladders swelling. Virions produced in nucleus of infected cells which hypertrophy without occlusion body production. 2001). 1995. for example through the automatic nervous on digestive organs. van Hulten. 2006). Thailand and China Taipei. but analysis of WSSV DNA sequences has shown that is not related to baculoviruses (van Hulten et al. inactivated vaccine (chemical / biological) and immunization (immersion and injection). et al. VNN infected fish usually show neurological disorders condition associated with central nerves system and retina strong vacuolization (damage) (R. and Alday-Sanz. VNN penetrate nasal epithelium via olfactory nerves and bladder.107 bp (GenBank Accession No.. as well as through the sensor and is associated with motor nerve in skin epithelium (Dennis et al.. WSD causative Agent is white spot syndrome virus (WSSV) or white spot virus (WSV). 292. 1998.. Yang et al. and attacked olfactory ears. and a trilaminar envelope (Wang. is a double strand DNA virus (dsDNA) recently established by the ICTV to own a new genus Whispovirus. Virions have a large (80-120 x 250380 nm). Grouper in grow-out phase can be infected by VNN with pernasal changes. Naturally. 1996. 2006).287 bp (GenBank Accession No. 2001). In early reports.5. Flegel.

0 mm (Mahardika et al. Hepatopancreas. 1996: Wang et al. Sudha et al. but it become red for adult shrimp (Mahardika et al.. (1998) states that if the shrimp were WSSV infected but have not found signs of white spots. Histopathologic specific changes infectious WSSV was the finding of pathological changes in cells. hepatopancreas and intestine.largely due to several small insertions and one large deletion (12 KBP). white spots or patches on shrimp carapace has not been found in samples collected.5-3. there is one that will join the nucleosome origin of replication (Wang et al. yellow tail shrimp (P. Kasornchandra et al.. 2004).. a total of 531 putative open reading frames (ORFs) were identified by sequence analysis. 1996). In field observation (diagnosis).. 2004). The virus has the form of virions rod-shaped particles with a size of 305 x 127 30 11 nm.. size. in fifth and sixth segments of the last abdominal and spread throughout his body cuticle (Wang et al. 1998 and Peng et al. In morphology.. In the acute phase. 1996). prospective parent / size of consumption (subadult) and shrimp (adult). 1997b).. this virus is known as White Spot Disease (Mahardika et al. The observation showed that shrimp cultivation in these areas has been attacked by WSSV on the stage of PL. The attack target organs which can be use as attack indicator of such as gill cells. 2004). 1994) or Penaeid Rod-shaped DNA virus (PRDV) also called Penaeid acute viremia (PAV) (Inouye et al. Sudha et al. which caused by viral infection. While in Taiwan. kuruma shrimp (P. cellular pathology and nucleic acids. which has low tissue infection so that the white spots and redness of shrimp was not visible. These changes are called Inclusion Body (Alifudin et. and often swim on one side only. 2004).. swimming is not directed. In accordance with the genome size of 300 kb. penicillatus) and shrimp greasyback (Metapanaeus ensis) (Kasornchandra and Boonyaratpalin. Shrimps which infected by this disease will die in short period (Ministry of Maritime Affairs and Fisheries. Rod shapes called nuclear japonicas virus or RV-PJ (Inouye et al. Sudha et al. Lightner (1996) states that the WSSV caused by SEMBV viruses which classified as DNA (Dioxyribonucleic Acid) viruses and rod-shaped (Bacilliform)... (1998) states that the WSSV-infected shrimp will changes the experience in behavior such reduced swimming activity. in Japan a virus that attacks P. WSSV (Pm NoBllType) are grouped on Non-occluded Baculovirus. 2004). 181 Thirty-six ORFs were identified by screening and sequencing of a cDNA. Inclusion Body observed a swelling of cell nucleus Page 9 of 21 . and in the nucleus. there are white patches on the carapace with a diameter of 0. intestine and gills cells which attacked by WSSV disease characterized by damaged core hipertopi (eosinophilic hypertrophy) and cell inclusion bodies. (1998) also said that death will occurred in 15-28 days. japonicus). categorized as type III (chronic). among which 181 ORFs likely to encode functional proteins. The Nomenclature of this virus is very diverse. WSSV disease was founded and identified in Taiwan in 1992 which caused mass mortality of tiger shrimp (Penaeus monodon). 1997a). and the first white spots appeared on cephalothorak. The shrimp also tend to schooling on the edge of the pond and swim to the surface. 1998). 1999). white spots on the carapace has become the common sign (Wang et al.. In Thailand it is known as viral diseases or systemic SEMBV Ectomesodermal or White Spot Syndrome Baculovirus Virus (WSSV) (Lightner. In WSSV cases. al. subfamily Nudibaculoviridae and family Baculoviridae (Mahardika et al. 2003).. Pathological changes such as abnormalities caused the development of existing and varion buildup inside cells. 1997a..

gills. lymphoid organs.. for example. 2006. lymphoid organ.. the researchers re-discovered antiWSSV antibody in chicken yolk egg. connective tissue. ranging from 5 to 20 percent.They are also testing chicken¶s serum after vaccination. glands and antenal haematopoetic. WSSV virus damages the stomach. "You do this by chickens vaccinating with WSSV virus with a certain dose." Three weeks pasca vaccination. 1997a).(hyperthropi cell nucleus) which are eosinophilic (reddish color) and move to the edge of cell nucleus. On research. Sri and her colleagues tested the Y-lg shrimp feed with a certain dose of infected white shrimp in ponds. Sri Murtini explains utilization Ig-Y research performed on a variety of bacterial diseases. Procambarus clarkii (Jha. antennal gland and haemocyte.. one week after the fourth vaccination. This feed test results proved Page 10 of 21 . Based on these. which is formulated in shrimp feed to control WSSV disease. Bogor Agricultural University (IPB). (2004). But a few scientists who examined disease caused by viruses in fish and shrimp.. The dose varied. Wang et al. Okti N Putri and Heru from Faculty of Veterinary Medicine at IPB is applying specific antibodies against WSSV7 on Y-yolk immunoglobulins (Ig-Y). 1998) and Pacifastus leniusculus (Jiravanichpaisal et al. both on terrestrial and marine animals. gill and nervous tissue (Wang et al. Chicken yolk egg which tested is proved contain Ig-Y is able to function as antibodies against WSSV virus. 1996). 2001) while Procambarus clarkii reaches 81% within 18 days after consuming a diet of shrimp (Penaeus monodon ) which positive WSSV (Wang et al. WSSV virus is able to spread through cannibalism among shrimp or through contaminated water (Chang et al. it have known that the WSSV virus against freshwater crayfish Pacifastus leniusculus result in total mortality on 10 days post infection (Jiravanichpaisal et al. Subsequently. This research conducted by Sri Murtini. Ig-Y is imunoglobulin or antibody proteins which will work to minimize the WSSV virus attacks and make this virus can not infect the host white shrimp. subkutikula. Changes in salinity did not affect WSSV virus. epithelial cells. Retno D Soejoedono. Inclusion Body can also be basofilik (colored blue) where the cell nucleus is very large in threshold to be broken (karyolisis) and it ended with cell lysis where cells nucleus out of the cell. Anti-WSSV antibody in yolk egg was tested by researchers from AGPT IPB. This is known in several studies that use freshwater crayfish Cherax quadricarinatus (Djauhari. Then through histopathologically observed. This research. According to Mahardika et al. The research team Faculty of Veterinary Medicine. researchers found anti WSSV antibody in chicken¶s serum.. by making eggs containing anti-Ig-Y WSSV for shrimp feed. immersion and injection.. beginning with the Ig-Y produced in chickens. the researchers conducted various researches in controlling this disease. develop a study to provide passive immunization with Y-immunoglobulin (Yolk immunoglobulin) in white shrimp. the cell degeneration occurs in the form of enlargement on the various tissues such as the mesodermal and ectodermal layer of skin. 2001) can be WSSV infected by oral. 1998). 2005).

but the community has a lot to use this vaccine. known as white spot ini. The seed-sized fish in this vaccination are 6 cm long. with 120 fishes. which were given Ig-Y 20%. hydrophila vaccines which is sold freely by BRPBAT Bogor. which 40 fishes were injected intraperitoneally. Different results seen in virus-infected shrimp and are not fed with Ig-Y. In ponds. Unfortunately. while for the non-vaccinated fish only reach 40%.1 Aeromonas hydrophila vaccine Aeromonas hydrophila vaccine use in field applications has yet to have any data.2. the vaccination carried out on other fish are estimated to remain healthy. In fish seed.1 ml/fish with bacterial density 107 cells/ml. if any fish which showed illness symptoms and otherwise ill based on laboratory test results. While shrimp fed with Ig-Y content of 5 and 10% getting better condition on the fifth day. for example. then the production of A. Shrimp like this die in a week. whether WSSV infection or due to other causes. From the PCR results concluded that the death of white shrimp which fed with anti-WSSV not find the virus. Vaccination is applied using vibrio vaccine polyvalent with densities 107 cells / ml Vibrio vaccination on tiger grouper had been done in January-March 2007. III. Efficacy laboratory-scale testing of 100 tilapia vaccinations with Aeromonas hydrophila vaccines by immersion results showed 90-100% survival rate after challenge test. so the vaccination result can not be described statistically. vaccine is applied in tiger grouper while for broodstock. Fish vaccinated by injection using a dose of 0. WSSV-infected shrimp condition began to improve after three days feeding this nutritious. Moreover the antibody titers of vaccinated fish reach 211 HAU/25 l which means an increase in immunity against Aeromonas hydrophila. both at the parent and the seed. From the research that has been done by BRPBAT Bogor. Fish vaccinated by immersion using diluted vaccine until the bacterial density into107 cells/ml. and then fish in Page 11 of 21 . Vaccination is applied polyvalent vibrio vaccine. 40 others are immersion and the last 40 vaccinated orally. in the implementation of vaccination has not been supported by the existence of a good recording system. Application in Central Sea Water Aquaculture Development (BBPBL) Lampung Vaccination Program in BBPBL Lampung has been running on grouper culture. The death shrimp then were tested by PCR to detect WSSV virus.2 Vibriosis vaccine application 3. 3. the research team concluded that the yolk egg containing Ig-Y that can be formulated in shrimp and capable of providing passive immunity of white shrimp WSSV virus-infected.1. Vaccination applied in BBPBL Lampung depending on fish condition report. It goal was to find out the causes of death.quite satisfactory. Efficacy test of Aeromonas hydrophila vaccines test currently only on laboratory-scale and will test in the field on 2011. Field Applications 3. is applied to giant grouper. Based on this study.

high Active swimming. symptoms that occur after vaccination should be continuously monitored. the fish condition must be really comfortable and healthy. high Do not swim actively. Appetite high appetite 13-14 Active. high Active swimming. Appetite high appetite Active swim. relative level of protection (Relative Percent Survival/RPS) and Mean Time to Death (MTD). it was obvious that after vaccination against disease.5 72. began there high appetite Active swimming. the fish in the gather in the primary primary vessel vessel Active. for vaccine application. Orally vaccinated fish was done by the fish fed with vaccine 3-5% per day for 10-15 consecutive days. These data also indicates the use of immersion as a way of vaccination is less effective because it raises stress on fish. Booster performed 7 days after the first vaccine. high appetite Page 12 of 21 . high appetite Active swimming. high Lethargy. high Appetite Active. Appetite Decreased appetite. Besides the parameters calculation. appetite. inactivity swim Lethargy. high Appetite Active. Parameter measuring the success of vaccination carried out with calculation of survival rate. Appetite increased Active. relative percent survival and Mean Time to Death reaches 100%. Thus. began to Weakness. do not swim actively gather gather 11-12 Active. it is aimed at monitoring fish condition during maintenance. appetite Appetite swim actively began to increase Active.5 72. Do not swim actively. Here are symptoms which appear: Day 1-3 Nonvaccinated Injection Immersion Oral Active.the soak for 15 minutes. loss of Appetite appetite. high Appetite Booster 80-10 Active. the fish resistances become much stronger were shown in increased survival rate. loss of Somewhat active. There is no appetite. Decreased appetite. high Appetite 4-5 6-7 Active. appetite Active swimming.5 RPS (%) 0 100 42.5 MTD (days) 6 ~ 9 14 From data obtained. Here are the results of the calculation parameters: Route of vaccination Non-vaccinated Injection Immersion Oral Survival rate (%) 0 100 42. high Began to eat.

Vaccines are made in isolation from the tiger grouper. 3. The symptoms were observed also as follows: Day 1-2 3-5 6-8 9-21 External symptoms during the challenge test The fish swam off. V. the decline in conditions such as fatigue and decreased appetite indicate side effects of vaccines in fish. Then these four isolates being test antibody titers and cross reaction to search for vaccine candidates. but after Koch's postulates test. and excessive mucus Fish started to move actively. tiger grouper were vaccinated with vibrio polyvalent vaccine (107 cells/ml) which was developed by BBPBAP Situbondo with dose 0. and V. 2010. fluvialis. To determine the effectiveness of the vaccine then performed challenge test.e. The bacterial isolates obtained from 29 isolates of bacteria. appetite had begun to exist.5 m with a density of 50 individuals per plot. gather.2. trachuri. lenders are normal. These four isolates are identified similar to V. After the acclimatization process.1 ml per fish via injection intraperitoneally using automatic injector. Applications in Situbondo Brackish Water Aquaculture Development Centre (Balai Besar Perikanan Budidaya Air Payau. But the recording of this vaccination does not exist.The symptoms which appear indicate that the fish in healthy condition. only obtained 14 isolates which have pathogenic properties.2. V. Usually these symptoms will begin to re-emerge about 2-3 months after vaccination. Vaccination applied to tiger grouper with 10 cm total length which kept on floating nets size of 1×1×1. booster being done with the same steps as in the initial injection. increased appetite Fish are active swimmers. lethargic and always quiet. alginolyticus. alginolyticus. Vaccination being done if there are grouper giant which showed any marks the emergence symptoms in reddish spots on fish abdomen. Acclimatization of these fish is less than a week before vaccination. Page 13 of 21 . BBPBAP) Situbondo has successfully created vibrio polyvalent vaccine to prevent vibriosis disease on 2008 to 2009. in which were identified similar to V. adult and larvae of Humpback grouper. giving high antibody titer against all vibrio isolates tested so that these serve as vaccine candidates. the mucus begins to decline. Pelagius with an average similarity level of 80%. has begun to respond Fish have started to swim on the surface with a swirling motion (whirling). After one week. The number of breeders which routinely vaccinated every three months is about 17 fish. Four of the most virulent isolates then being pathogenicity test and obtained the LD 50. and high appetite Vaccination to parent fish was applied to giant grouper. In the following year i. low appetite. The results of vaccination showed reduced levels of disease symptoms giant grouper incidence. From these tests is known that TKL-2 isolate. Fish vaccinated using a vibrio vaccine with dose of 1ml/fish via injection intraperitoneal. the vaccine is applied to tiger grouper cultivation floating net cage in BBPBAP Situbondo.

length and average weight of fish. namely streptococcal and vibrio vaccines. BBPBAP Situbondo actually has two vaccines types. should not be attacked by disease and health status examination should be performed prior to fish vaccination. Parent fish vaccination procedures: 1.From the vaccination applied. while non-vaccinated fish about 50%. obtained data of antibody titer values. Fish condition must be physically clean and fish also should be kept in free of pest water and fish diseases. Table antibody titer results Treatment Control Of vaccinated fish BO 2 6 B1 2 5 B2 2 5 B3 24 29 26 27 28 BO: Bleeding before vaccination B1: Bleeding a week after vaccination BO: Bleeding a week after vaccination BO: Bleeding the end of observation The antibody titers results showed that the vaccine tended to increase the antibody. Fish length and weight data also showed an increase in body weight of vaccinated fish at 21. 2. In field application. Survival rate obtained indicates that resistance of vaccinated fish higher than non-vaccinated fish against V. Here are the procedure adopted in BBPBAP Situbondo vaccination both on breeding and seed grouper: A. Broodstock selection Broodstock which to be vaccinated must be in good health. Stress can decrease health conditions so that vaccination of stress fish would cause fish illness because of the immune system weakened to receive the vaccine and this vaccine can actually cause fish Page 14 of 21 . Acclimatization and adaptation This process aims to minimize the stress level of fish to be vaccinated.165%. the streptococcal vaccine has not been done while vibrio vaccine has been applied to the broodstock and seed grouper. This shows that the V. Recently BBAP Situbondo have been vaccinating with vibrios as many as 20. These indicate that fish in a more healthy condition so that its appetite are also improving.69%.alginolyticus polyvalent vaccine can provide good protection. especially a high level of survival live. so that the recording of data as a result of vaccination continued to be monitored.000 grouper belonging to private and the surrounding communities. and survival rate. (2005) that vaccine can increase fish resistance. At the end of observation the average survival rate of vaccinated fish were reach 68. Vaccines use report in grouper showed that the vaccinated parent has diseases incidence which tend to be lower than prior to vaccination. alginolyticus during maintenance. This is in accordance with the opinion of Kamiso (1990) and Kamiso et al.

Seed Selection Seeds to be vaccinated must be in good health. In this process if there are parasites. Fish vaccination procedure 1. then fish should be cleaned and cleared of parasites attack. 5. Stress can decrease health conditions so that vaccination of stress fish would cause fish illness because of the immune system weakened to receive the vaccine and this vaccine can actually cause fish sick. 3. Maintenance Fish that have been vaccinated are kept in free of pest¶s water and fish diseases which both environmental conditions and water quality of the feed should be kept for 3-7 days. Vaccination can be done with injection. Then these fish are ready to sell or exaggerated. or oral (via feed). In this process if there are parasites. 3. intramuscular and intra-venous. 4. Seed booster is done about a week (7 days) after the first vaccination. Booster Repetition of the vaccine or booster aims to strengthen the fish immune system. Page 15 of 21 . Vaccination can be done with injection. or oral (via feed). Time and method of vaccination Vaccination of seed fish is done about 15-20 days after spawning. Vaccination is usually done via intramuscular injection. Acclimatization and adaptation This process aims to minimize the stress level of fish to be vaccinated. Breeding B. then fish should be cleaned and cleared of parasites attack. Fish condition must be physically clean and fish also should be kept in free of pests water and fish diseases. Time and method of vaccination Vaccination of parent carried out about two weeks before spawning. soak. 2. Seed vaccination is usually carried out through intraperitoneal between two fin fish 4. soak. Maintenance Fish that have been vaccinated are kept in free of pests water and fish diseases in which both environmental conditions and water quality of the feed should be kept.sick. in order to produce seeds that have been brought immunity from its mother so as not susceptible to disease and can be specific pathogenic free. For delivery by injection can be done through intraperitoneal. 5. should not be attacked by disease and health status examination should be performed prior to the vaccination of fish.

the other observation is that the symptoms occur. Asian seabass of the seed with length and weight and different density conditions were vaccinated with vibrio polyvalent vaccine used best dose from laboratory scale test results through the feed. The observation during five months activities are as follows: A. During the challenge test fish mortality rates and time of death occurrence will be observed. Feed plus vaccine was given during three consecutive days to test fish. Fish Length No. Two weeks after the booster antibody titers were observed. The booster vaccination conducted after one week with the same method and dose of vaccination. Both vaccination and booster conducted over three consecutive days via feed. these fish moved in floating nets mesh size of 3×3×3 m3 in density 1000 fish/net for further observation during five months (150 days of research). while as a control given the same diet without the vaccine. 1 2 3 4 Sampling Date March 15 April 16 May 16 June 16 Observations of Fish Length (cm) Test fish 22 31 39 47 Control 23 30 38 45 Page 16 of 21 . which the feed were given chicken white egg as a binder of feed and vaccine. Preparation vaccines in food is done by spraying the 1000 ml vaccine into 2000 g of feed pellets which had been sprinkled with grains 6 white egg as binder vaccine. One week after booster.3. challenge test carried out by intramuscular with a dose of 106 -07 CFU/ml and subsequently maintained in the aquarium with good condition.3. Fish Weight No. Application in Batam Institute for Marine Aquaculture Batam has applied polyvalent Vibrio vaccine in February until June 2010. Maintenance begins on day 4 to day 14. The vaccine used was polyvalent Vibrio vaccine with densities 107 cells/fish. Vaccination is done every day for the last 3 (three) days of vaccination via feed. which will be a booster with the same treatment.2. To test the vaccine efficacy. 1 2 3 4 Sampling Date March 15 April 16 May 16 June 16 Observations Fish Weight (grams) Test fish 180 230 300 410 Control 182 223 287 395 B. The vaccination method in laboratory scale tests were orally through feed.

At the observation of fish final length reach 47 cm.1 cm and 410 g. which means daily weight rate gain was Page 17 of 21 ¢ ¡  . Survival rate Treatment Non-vaccinated Vaccinations Survival rate (%) 83 92 Based on the above data. E.247 cm length/head/day. It is better when compared with the control fish with final length 46 cm which means daily increasing rate was 0. Fluctuations in Total Bacteria General (TB ) and Total Bacteria Vi i (TB ) during the observation. its means that during the 150 days of maintenance there is increase gain 0.240 1.3 289.450 1. Observations of vaccinated fish blood.233 cm/head/day. which is fish survival rate at the end of test is 92% and 83% for control. 32.2 Haemoglobin (g / dl Leucocytes (cells / Hematocrit (/ Thrombocyte (/ liter) liter) liter) liter) 501. 1 2 3 4 5 Carcass anal sis Fish samples Test fish Control 5.C Blood Anal i No. average fish¶s length and weight in final sequentially for the test is 35.490 27.8 cm and 395 g in the control fish.62 6.9 487.9 233. The average weight at the end of maintenance period on the test fish is 410 grams. Likewise fish weight increasing rate. showed improve the immune response based on increasing the number of leucocytes concentration and haematocrite.89 Erythrocytes (million / D. it is known that vaccination gives better results thannon-vaccinated fish.290 32.

Based on common bacteria and Vibrio total number. Furthermore. Administration vaccine through injection used at the first step of this research. which at the same time teaching staff at the Aquaculture Department of Fisheries and Marine Science Faculty of Bogor Agricultural University has made a DNA vaccine for KHV.. Results of reverse transcriptase PCR analysis showed that the gene contained by the vaccine expressed in the vaccinated fish which means that the vaccine is active.37 g/head/day.290 (j / liter) and haematocrite was 32. DNA vaccine is an alternative effective and safe vaccine. TBU range from 20 to 4. Through research sponsored by the Department of Marine and Fisheries of West Java Province (year 2008) and Doctorate Grant research program by the SPS-IPB (year 2009).Si which is a student of Veterinary Science Doctoral PS-SPS IPB.47 g/head/day. Because of the ability of DNA vaccine protection large enough. This compares favorably with the daily weight growth rate of control fish were 2.25 x 103 and Total Bacteria Vibrio 20 to 6. This can happen because vaccination is one effort to deal with animals (including fish) diseases by giving vaccine into the animal's body in order to have resistance from disease attack. In subsequent studies will be developed a simple method to mass applied in the field. But vaccination showed effectiveness in providing resistance to disease attack.Pi. the vaccination method development for applications in the field need to be implemented. This means that the immune system that you have on vaccinated fish was better than the control fish. M. This is evidenced by relatively high survival rate of 92% versus 83% with control fish.3 (j / liter). While the survival rate for vaccinated fish about 92% better than control fish was 83%. Non-vaccinated fish that are before the challenge test are all done to death.2. From proximate analyze and blood profiles in vaccinated and control fish.3 KHV Vaccine According to Alimuddin (2010). which until now the problem is not getting the correct solution (Alimuddin.490 (j / liter) and haematocrite 27. attenuated virus vaccines such as vaccines above has been available. This research is expected to contribute to prevent KHV infection in fish farming that have occurred in Indonesia since 2002. 3. weekly fluctuations in water are very high. such as through live food and feed. While the control fish leucocytes about 487. This shows that DNA vaccines are very effective to cope with KHV virus attack. 2010).8 x 102 is a sufficient condition alarming occurrence of both primary and secondary infection in asian seabass cultivation. Sri Nuryati. it is known that the vaccinated fish have better nutritional content compared with control fish and especially the content of leucocytes and haematocrite blood profile showed concentrations increase in vaccinated fish leucocytes contain about 501. Page 18 of 21 . through laboratory-scale challenge test the vaccine can increase survival of common carp more than 95% for one month after vaccination. however the price is still considered to be very expensive and from the testing that some indication of the virus is still virulent.9 (j / liter). S.

Although the application via intramuscular injection is a method that can be considered in vaccination. and (4) WSSV challenge test. with the same amount of fish given 15 mUL inactive vaccine by soaking treatment A (one hour). especially to increase the fish's immune system against various bacterial and viral diseases. 3. 360 juvenile challenged with VNN during six hours immersion. 3. Further research and monitoring needed for vaccination technology develop included new vaccines produce and the administered methods in order to obtain an effective vaccine. (3) Vaccines application by immersion method. After 10. Parameters observed included the highest survival rate of giant tiger prawn (Penaeus monodon) post-larvae in treatments. efficient.5 WSSV vaccine WSSV vaccine was applicated by the The Research Institute for Brackish Water Aquaculture (Balai Riset Perikanan Budidaya Air Payau. In second stage. KHV. There are two stages of the experiment. Various researches and its application in Indonesia showed some vaccines such as Aeromonas hydrophilla. Page 19 of 21 . The results of VNN challenge in first stage show the treatment of 15 ml/L gives survival rate amounted to 86.4 VNN Vaccine Research was conducted by Gondol Research Institute for Mariculture to know the effect of different doses and duration of immersion about humpback grouper juvenile (Cromileptus altivelis) immune against VNN infection. and observed clinical symptoms and mortality. and VNN have been able to clinically test in preventing disease caused by the target organism.2 ml vaccine per fish intraperitoneally. and 30 days post vaccination 10 juvenile each challenged with VNN for 6 hours. and economical and applicable. Vaccination dose of 15 ml/L with 3-hour soaking can improve the humpback juvenile immunity against VNN infection. but the development of applications with other methods should be developed for example by immersion or by mixing with a feed with consideration for environmental security. The result of second stage is on immersion treatment for 3 hours gives survival rate after challenge about 86. and D (0 hours / control). Vaccine dose was obtained by humpback grouper seed treatment with injection of 0. Then the fish were reared for two weeks. C (3 hours). But apparently there were no significant differences for all treatments. 20. Conclusion Vaccines application has many advantages. Another report on WSSV vaccine application is not yet known. (2) WSSV LC50 Test. IV. Vibriosis.67% after 25 days post-vaccination. B (2 hours). Further observations should be made as in first stage. first stage. VNN vaccine applications in grouper seed after 30 days VNN challenged test can reduce mortality also increase survival rate and titer of humpback grouper seed. BRPBAP) Maros which consists of several stages: (1) Production of WSSV vaccine.67% after 20 days post-vaccination.

. Purifikasi dan Immunogenitas Protein Outer Membran Vibrio alginolyticus pada Ikan Kerapu Macan (Epinephelus fuscogutatus). Efikasi Vaksin Debris Sel Aeromonas hydrophila Secara Suntik Dengan Variasi Cara Booster Pada Lele Dumbo (Clarias gariepinus Burchell). Page 20 of 21 . dan Ngurah P.wordpress. YD.S. Unpublish. Pencegahan Infeksi Viral Nerveous Necrosis pada Benih Ikan Kerapu Bebek (Cromileptus altivelis). Sumanta. 2000. 1816-1820 Qian. R. Chu. www. Pengaruh Jenis Vaksin dan Konsentrasi Vitamin C terhadap Sintasan Pasca Larva Udang Windu yang dipapar dengan WSSV. HJ.156. Disertasi. Vaksin DNA untuk Virus KHV yang Menyerang Ikan Mas dan Koi. Murtini S. 2004. Casessar. Jurnal Perikanan. A. Identification of Vaccine Candidates Against Serogroup B Meningococcus by Whole-genome Sequencing. W.id/basic data/index. Maros. 2007. Mao. Immunology. Enginering report. 2010. Expression.H. L. Haryanti. www. Isolasi. Mulia..lppm.J. Ambariyanto. dan Okti. Yudiati. Z.39.rumahkucing.. 2006. Sin. Pengembangan Vaksin DNA Penyandi Glikoprotein Virus KHV (Koi Herpes Virus) menggunakan Isolat Lokal. Characterization and Immunogenicity of a Major Outer Membrane Protein from Vibrio alginolyticus.References Alimuddin. Kuby. Sri Nuryati. Biophys. Aplikasi Antibodi Spesifik terhadap WSSV 7 pada kuning telur Immunoglobulin-Y (Ig-Y) yang diformulasikan dalam pakan Udang Untuk Mengendalikan WSSV. 2009. Freeman and Company. 194-200 Santika A.com/2010/07/15/whitespot-syndrome-virus Nuryati. 2007. Taslihan. Uji Vaksin DNA anti KHV Skala Laboratorium. R. and Triyanto. BBRBPL-Gondol Bali. Biologi 3 (3): 145 . Sri. Acta Biochim. Jurnal Penelitian Ilmiah Vol 10 No 1 Desrina. N. http://ballstenford. Muliani dan Madeali.. London. Desrina. Taylor and Franchis Ltd. J. Scarlato. Aplikasi Antigen Murni sebagai Vaksin untuk Pengendalian Penyakit Vibriosis pada Ikan Kerapu. Situmeang.brkp.. 2004. W. 1985.P.com Atmowarsono.go. 2009. V. 1992. and Masignani. Sembiring.. D.php?com=riset&taskview&id=694&PHPSessiD Kabata. Parasiter and Diseases of Fish Cultured in tropic. V. Pizza. Laporan Riset Unggulan terpadu Bidang Pertanian dan Pangan. BS. Triyanto. D. Balai Riset Perikanan Air Payau.H. Pratiwi. lppmipb. Science 287. E.. 2010. Z.Y. New York. Retno.dkp. 2010.M.

Fish Immune System: A cross between innate and adaptive responses.S. 12503-12505 Ward. R. The Development of Bacterial Vaccines for Fish. and G. Press. Robert (Ed. P.A. Proc.. Paper presented in the Symposium of Fish Vaccination. no. Balasch C. Pengelolaan Usaha Pembenihan Ikan Mas. Immunologia Vol 22. Acad. Agglutinating Anti bodies Production in Eel (Anguilla japonica) Inoculated with Aeromonas hydrophila Antigen. London Page 21 of 21 . In R..) Microbial Diseases of Fish. Suseno D..N. Sci. Comparative Genome Analysis Of The Alpha-Proteobacteria : Relationships Between Plant and Animal Pathogens and Host Specifity. 99. Acad. Mckenzie S. Natl. Chen.D. U.3: 277-286 Tsolis. 1982.M. Penebar Swadaya.Song.J. 2002. Y. Kon. OIE Fish Diseases Commission Paris Thesis. 2004. Jakarta. Auburn University. 1984. Tort L. S. 2003..H. PT.