Imunohistokimia Patogenitas Viral Nervous Necrosis Isolat Lapang Bali yang Diinfeksikan pada Kerapu Macan Budidaya

Di Indonesia dilaporkan bahwa VNN (Viral Nervous Necrosis) telah menyerang sebagian besar budidaya ikan kerapu dengan tingkat kematian 100%. Untuk mencegah penyebaran penyakit VNN pada kerapu yang dilalulintaskan Balai Karantina Ikan Pengendalian Mutu dan Keamanan Hasil Perikanan kelas I Denpasar mengembangkan pemeriksaan imunohistokimia untuk mengetahui tingkat patogenitas VNN sebagai dasar penanggulangan dan pencegahan penyakit VNN di wilayah Bali. Kerapu macan berukuran 150 g - 300 g sebanyak 50 ekor diaklimatisasi, sepuluh ekor kerapu sebagai kontrol, 40 ekor diinjeksi dengan inokulum VNN (isolat Bali) konsentrasi 101,5 yang dipelihara tanpa siklus pergantian air. Pengamatan gejala klinis dan pengambilan sampel organ dilakukan 12 jam pasca infeksi dan berturut-turut setiap 12 jam. Pengambilan organ digunakan untuk pemeriksaan imunohistokimia (streptavidin-biotin) dan uji konfirmasi menggunakan pemeriksaa RT- PCR kit IQ-2000 VNN. Hasil imunohistokimia pada 24 jam pertama hanya menyerang mata dan otak, 36 jam pasca infeksi virus penyebab VNN terdapat di organ jantung, 48 jam pasca infeksi virus penyebab VNN terdapat di organ hati dan limpa yang diakhiri pada 60 jam pasca infeksi virus penyebab VNN berada di organ ginjal. Distribusi VNN yang luas pada seluruh organ tubuh menunjukkan bahwa viraemia menjadi faktor penting dalam patogenesis infeksi penyakit VNN.Kata kunci: VNN, patogenitas, kerapu macan, imunohistokimia (Imunohistochemistry of Pathogenicity Viral Nervous Necrosis Bali Field Isolate are Infected in Tiger Grouper Marine Culture)In Indonesia was reported VNN (Viral Nervous Necrosis) has invaded most of grouper culture with a mortality rate of 100%. To prevent the spread of VNN in groupers export from Balai Karantina Ikan Pengendalian Mutu dan Keamanan Hasil Perikanan kelas I Denpasar, it develop immunohistochemistry to determine the level of pathogenicity VNN as a basis for prevention and disease prevention VNN in Bali. Tiger grouper sized 150 - 300 g as much as 50 animals acclimatized, ten head of grouper as a control, 40 tails were injected with an inoculum concentration of 101.5 VNN are preserved without change of the water cycle. Observation of clinical symptoms and organ sampling performed 12 hours post-infection and successively every 12 hours, making the organ used for immunohistochemistry (streptavidin-biotin) and a confirmatory test using RT - PCR kit examination of the IQ -2000 VNN. Immunohistochemical results on the first 24 hours only attacks the eyes and brain, 36 hours post-infection the virus that was found in cardiac, 48 hours post-infection the virus that VNN contained in the liver and spleen were terminated at 57 hours post-infection the virus in the kidney. Spread of VNN distribution in all organs showed that viraemia is an important factor in the pathogenesis of infectious diseases VNN.Keywords: VNN, immunohistochemistry, tiger grouper, patogenita

Analysis of Traffic Volume and Frequency of Vannamei Shrimp (Litopenaeus vannamei) Shipments Based on a Certification Approach

Volume and frequency of shipping are crucial factors that impact the profitability and sustainability of the vannamei shrimp industry. This study aims to determine the volume of vannamei shrimp shipping traffic in Padangbai, Karangasem, Bali during the period 2019-2021 and the relationship between the volume of vannamei shrimp shipping traffic and shipping times during that period in the region. Data were obtained from fish quarantine stations, quality control and fishery product safety in Padangbai, Karangasem, Bali. The analytical method is carried out in a comparative descriptive manner which is presented through graphs that have been processed in the MS program. Excel while to find out the relationship between volume and frequency of shipping was analyzed using regression. The volume of traffic for shipping vannamei shrimp out of the Padangbai area, Karangasem, Bali was highest in 2021 (652,745,431 Juvenils) and lowest in 2019 (435,740,234 Juvenils). Data on the frequency of traffic was highest in 2020 (383 times) and lowest in 2019 (268 times). Based on the data provided, it can be seen that the average total vannamei shrimp shipment during the 2019-2021 period was around 4,702 x 104 juveniles per month. There were fluctuations in shipment volume from month to month, where in February, March, August, September and December the highest shipments were recorded of more than 5,000 x 104 juveniles, while in other months the lowest shipments were recorded of 2,929 x 104 juveniles in June. Based on these data, it can be concluded that the volume of vannamei shrimp shipments from Padangbai, Karangasem, Bali is quite stable with relatively small fluctuations. Even though there are fluctuations in shipping volume from month to month, overall, the volume of vannamei shrimp shipments from Padangbai, Karangasem, Bali is quite high

Immunocytochemistry Streptavidin Biotin: Early Detection of Viral Nervous Necrosis Virus in the mucous of the Ikan Kerapu Macan(Epinephelus fuscoguttatus)

Kerapu macan  (Epinephelus fuscoguttatus)    is  one of the  kerapu  fishes  that  has been successfully  bred by the  farmers.  The  survival  rate  of  the  fish  reaches  up  to  the  40%  at  the  fish  hatchery  of  the  home  industry  of  in  Bali. The disease in which may cause significant mortality in kerapu fish, especially for the larvae and juveniles is viral nervous necrosis (VNN). We therefore developed and applied the immunological diagnostic approach of immunocytochemistry technique of streptavidin biotin (SB) for early detection of  VNN. Mucous samples of VNN  infected-  kerapu  fish  in  vivo  were   firstly  detected  by  reverse  transcriptase-polymerase  chain  reaction  (R T -PCR)  for  the  presence  of  DNA  VNN.  If  it  is  RT -PCR-positive  VNN,  the  mucous  samples  were  then  tested  by  SB. Results of the present study indicated that the VNN virus could be detected with SB technique within only 24 hours post infection.  It  was concluded that  rapid  and accurate SB technique  is  suitable and appropriate to  be applied  for  routine  control  and  prevention  national  program  in  the  Fish  Quarantine  for  Indonesia  because  no  need of  fish  sacrification,  and  scientific,  law  and  international    accepted,  and  even  no  hazardous  environmental contamination.Keywords:  kerapu  macan  (Epinephelus  fuscoguttatus),VNN,  mucous,  SB,  early  detectio

Imunositokimia Streptavidin Biotin: Deteksi Dini Viral Nervous Necrosis Virus pada Lendir Ikan Kerapu Macan (Epinephelus fuscoguttatus) = Immunocytochemistry Streptavidin Biotin: Early Detection of Viral Nervous ...

Kerapu macan (Epinephelus fuscoguttatus) merupakan salah satu diantara ikan kenipu yang berhasil dibudidayakan oleh petani petambak dan tingkat keberhasilan mencapai 40% pada penetasan kerapu industri skala rumah tangga di Bali. Virus viral nervous necrosis (VNN) merupakan penyebab kematian massal ikan kerapu, terutama larva dan juvenil. Pada penelitian ini, dikemba.vgkandan diaplikasikan uji imunositokimia streptavidin biotin (SB) untuk diagnosis dini VNN. Sampel (lendir) ikan kerapu yang diinfeksi virus VNN in vivo, diuji reverse transcriptase-polymerase chain reaction (RT-PCR).Lendir yang positifDNA VNN dengan RT-PCR,selanjutnya diuji SB. Hasil penelitian ini membuktikan, bahwa virus VNN dapat dideteksi pada lendir ikan kerapu macan tersebut yang terinfeksi virus VNN 24 jam sebelumnya. Disimpulkan, bahwa uji SB yang cepat dan akurat adalah tepat dan cocokuntuk diaplikasikan dalam rangka program rutin kontrol dan pencegahan VNN di Karantina Ikan Indonesia karena dapat dilakukan tanpa mematikan ikan, diterima secara ilmiah, hukum Dan internasional, dan bahkan tidak mencemari lingkungan hidup. Kerapu macan (Epinephelusfuscoguttatus) is one of the kerapu fishes that has been successfully bred by the farmers. The survival rate of the fish reaches up to the 40% at the fish hatchery of the home industry of in Bali. , The disease in which may cause significant mortality in kerapu fish, especially for the larvae and juveniles is viral nervous necrosis (VNN). We therefore developed and applied the immunological diagnostic approach of immunocytochemistry technique of streptavidin biotin (SB) for early detection of VNN. Mucous samples of VNN infected-kerapu fish in vivo were firstly detectedby reverse transcriptase-polymerase chain reaction (RT- PCR) for the presence of DNAVNN. If it is RT-PCR-positive VNN, the mucous samples were then tested by SB. Results of the present study indicated that the VNN virus could be detected with SB technique within only 24 hours post infection. It was concluded that rapid and accurate SB technique is suitable and appropriate to be applied for routine control and prevention national program in the Fish Quarantine for Indon6siabecause no need of fish sacrification, and scientific, law and international accepted, and even no hazardous environmental contamination

Examination of Viral Nervous Necrosis Virus in The Water Sample of The Tiger Grouper (Epinephelus fuscoguttatus) with Immunocytochemistry of Streptavidin Biotin

Abstract         One of potencies of marine waters that is developed, and starts to have the international market is grouper fish. Tiger grouper culture can not be separated from disease factors that may attack and destroy the cultivation. One of the important diseases that can cause mass mortality in fish, especially in grouper larvae and juveniles stadium is viral nervous necrosis (VNN). Examination by using immunocytochemical method of streptavidin biotin (SB) undertaken to develop early detection of VNN in order to be applied in preventing spread of the VNN disease in seeds of tiger groupers that are already distributed worldwide in Indonesia. And also, this SB methodology can also be applied to the tiger groupers without any injuring or even killing of the fish for the purpose of sampling. In the present study, the preliminary test was done to determine the infectivity level of the inoculum (VNN virus) owned by the Office of Fish Quarantine, Quality Control, and Fishery Safety Class I Denpasar, Bali. Whereas, the main test was done by collecting water sample from tiger grouper fish farming that was infected with VNN. The water sample was examined by using  immunocytochemical method of SB after being confirmed by RT-PCR-positive test that was previously purified. Based on both SB and PCR tests having VNN positive results, it is concluded that  the immunocytochemical method of SB could be applied as early, rapid and accurate detection of VNN  in only a 24 hours post infection by using water as the sample. Keywords: VNN, water sample, streptavidin biotin, RT-PCR, tiger grouper

Infection of Infectious Spleen and Kidney Necrosis Virus and Viral Nervouse Necrosis in Seawater Fish Identifed Using Multiplex Polymerase Chain Reaction

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.