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Genetic Heterogenity Profile of Penaeus monodon Broodstock F1 Revealed by Mitochondria DNA-RFLP and RAPD Prastowo, Bambang Widyo; Rahardianti, Rahayu; Nur, Evi Maftuti; Taslihan, Arief
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (172.63 KB)

Abstract

The genetic heterogeneity of Penaeus monodon broodstock F1 was evaluated using Restriction Fragment LengthPolymorphism (RFLP-mtDNA) and Random Amplified Polymorphic DNA (RAPD) analysis. The RFLP analysis wasconducted by amplifying 16SrDNA region and digested with restriction enzyme Nde II. According to the RFLPanalysis, heterogeneity value of P. monodon F1 broodstock population is 0,0422; male F1 population is 0,0613 andfemale F1 population is 0,1252. The primer OPA2 was used in RAPD analysis. According to the RAPD analysis,heterogeneity value of P. monodon F1 broodstock population is 0,0417; male F1 population is 0,0653 and female F1population is 0,1104. The results of this research showed that either RFLP or RAPD can be used as a family specificmarker for Penaeus monodon.Key words : Penaeus monodon, RFLP, RAPD, heterogeneity, genetic marker
THE PREVALENCE AND RISK FACTORS OF WHITE SPOT SYNDROME VIRUS IN TIGER SHRIMP AT TRADITIONAL PONDS (PREVALENSI DAN FAKTOR RISIKO WHITE SPOT SYNDROME VIRUS PADA TAMBAK UDANG WINDU TRADISIONAL) Taslihan, Arief; Sumiarto, Bambang; Nitimulyo, Kamiso H.
Jurnal Veteriner Vol 15 No 3 (2014)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

A research has been conducted to determine the status of the White Spot Syndrome Virus (WSSV)epidemic on traditional shrimp ponds in Demak regency. A cross-sectional research method was selected,by calculating samples using statistical calculation based on the assumption of 70% prevalence rate anderror (p<10%). Additionally, this research also determined the risk factors in the onset of WSSV shrimppond. Surveyed areas included Sayung and Bonang, subdistrict in Demak district, because the area areactively engaged with shrimp farming. A total of 90 traditional tiger shrimp ponds were sampled in thisstudy. The results showed that the prevalence of WSSV infected farms was 65.6% (Confident Interval,CI=55.7%-75.4%). Factors that affected high prevalence of WSSV were screening of seed (Coefficient=1.517,P<0.049), pond size (coefficient=0.0002, P<0.0089) and drying ponds (coefficient=3.756, P<0.001). Factorsthat affected the production of shrimp in ponds were analysed using multivariate regression analysis.Based on the analysis, some factors had an important role in production, i.e. : stadia of shrimp seed(coefficient=19.091, P<0.19), water depth in pond (coefficient=1.343, P<0.0024) and WSSV-infected shrimp(coefficient=-44.369, P<0.001). It was concluded that WSSV prevalence in studied area was 65.6%, and itconsidered as hyperendemic. Factors affected the WSSV prevalence were selection of seed, pond?s size anddrying ponds. Risk factors affected shrimp production in the pond were stocking seed stadia, whereasWSSV-infection of shrimp was negatively affected factor.
Isolasi, Purifikasi dan Immunogenitas Protein Outer Membran Vibrio Alginolyticus pada Ikan Kerapu Macan (Epinephelus fuscoguttatus) Desrina, Desrina; Taslihan, Arief; Ambariyanto, Ambariyanto; Yudiati, Ervia; Casessar, Yulius Docang; Sumanta, Raden Bagus Sugio; Triyanto, Triyanto; Situmeang, Hotnida Junita; Sembiring, Langkah
Jurnal Perikanan Universitas Gadjah Mada Vol 9, No 1 (2007)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jfs.57

Abstract

The main objectives of this research were to isolate, purify and determine the immunogenicity of the outer membrane protein of V. alginolyticus. The outer membrane protein was isolated by sonication, sodium dodecyl sulphate-polyacrylamide agarose gel electrophoresis (SDS-PAGE) and purified by electroelusion method. Four outer membrane proteins were obtained: namely 32.0; 37.83; 64.13 and 73.43 kDa. Its immunogenicity in grouper Epinephelus fuscoguttatus was compared to that of bacterin of the same isolate. The immunogenicity test was conducted by intra peritoneal injection method. Each protein was dissolved in sterile Phospate Buffer Saline (PBS) and Freund’s Complete Adjuvant (FCA) (1:1) and injected 5 µg/ fish (fish weight 10-15 g). Bacterin (106, 107 and 108 cells/ml) was prepared in the same manner and injected at dosage of 0,2 ml/fish. Control fish were injected with 0.2 ml sterile PBS (pH 7.2). Booster was done a week later by injecting protein or bacterin with the Freund’s Incomplete Adjuvant (FIA). The agglutination test of antibody produced recognized cell surface protein of the whole cell of V. alginolyticus. Outer membrane protein 73.43 kDa was more immunogenic than the rest of proteins and bacterin, based on agglutinating antibodi titer.
CLUSTER MODEL FOR EXTENSIVE GIANT TIGER SHRIMP (Penaeus monodon Fab.) TO PREVENT TRANSMISSION OF WHITE SPOT SYNDROME VIRUS Taslihan, Arief; Callinan, Richard; Torribio, Jenny-Ann; Sumiarto, Bambang; Nitimulyo, Kamiso Handoyo
Indonesian Aquaculture Journal Vol 10, No 1 (2015): (June 2015)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (76.288 KB) | DOI: 10.15578/iaj.10.1.2015.65-70

Abstract

White spot syndrome virus (WSSV) has become epidemic in Indonesia and affecting shrimp aquaculture interm of its production. White spot syndrome virus is transmitted from one to other ponds, through crustacean, included planktonic copepode as carrier for WSSV and through water from affected shrimp pond. A cluster model, consist of shrimp grow out ponds surrounded by non-shrimp pond as a role of biosecurity has been developed. The model aimed to prevent white spot virus transmission in extensive giant tiger shrimp pond. The study was conducted in two sites at Demak District, Central Java Province. As the treatment, a cluster consist of three shrimp ponds in site I, and two shrimp ponds in site II, each was surrounded by buffer ponds rearing only finfish. As the control, five extensive shrimp grow out ponds in site I and three shrimp grow out ponds in site II, with shrimp pond has neither applied biosecurity nor surrounded by non-shrimp pond as biosecurity as well considered as control ponds. The results found that treatment of cluster shrimp ponds surrounded by non-shrimp ponds could hold shrimp at duration of culture in the grow out pond (DOC) 105.6±4.5 days significantly much longer than that of control that harvested at 60.9±16.0 days due to WSSV outbreak. Survival rate in trial ponds was 77.6±3.6%, significantly higher than that of control at 22.6±15.8%. Shrimp production in treatment ponds has total production of 425.1±146.6 kg/ha significantly higher than that of control that could only produced 54.5±47.6 kg/ha. Implementation of Better Management Practices (BMP) by arranging shrimp ponds in cluster and surrounding by non-shrimp ponds proven effectively prevent WSSV transmission from traditional shrimp ponds in surrounding area.
PROBLEM SOLVING PENYAKIT DI PEMBENIHAN UDANG Murdjani, Mohammad; Taslihan, Arief
Media Akuakultur Vol 1, No 1 (2006): (April 2006)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (831.409 KB) | DOI: 10.15578/ma.1.1.2006.19-24

Abstract

Dalam era pasar global, pemasaran produk ke pasar internasional harus memenuhi beberapa kriteria, terutama adalah penampilan bagus, tidak menunjukkan adanya tanda infeksi penyakit, dan tidak mengandung residu baik antibiotika maupun pestisida
Genetic Heterogenity Profile of Penaeus monodon Broodstock F1 Revealed by Mitochondria DNA-RFLP and RAPD Prastowo, Bambang Widyo; Rahardianti, Rahayu; Nur, Evi Maftuti; Taslihan, Arief
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (172.63 KB) | DOI: 10.22146/ijbiotech.7769

Abstract

The genetic heterogeneity of Penaeus monodon broodstock F1 was evaluated using Restriction Fragment LengthPolymorphism (RFLP-mtDNA) and Random Amplified Polymorphic DNA (RAPD) analysis. The RFLP analysis wasconducted by amplifying 16SrDNA region and digested with restriction enzyme Nde II. According to the RFLPanalysis, heterogeneity value of P. monodon F1 broodstock population is 0,0422; male F1 population is 0,0613 andfemale F1 population is 0,1252. The primer OPA2 was used in RAPD analysis. According to the RAPD analysis,heterogeneity value of P. monodon F1 broodstock population is 0,0417; male F1 population is 0,0653 and female F1population is 0,1104. The results of this research showed that either RFLP or RAPD can be used as a family specificmarker for Penaeus monodon.Key words : Penaeus monodon, RFLP, RAPD, heterogeneity, genetic marker
PENGARUH DOSIS TERHADAP EFEKTIFITAS VAKSIN POM VIBRIO ALGINOLYTICUS 74 KDA PADA IKAN KERAPU MACAN EPINEPHELUS FUSCOGUTTATUS Desrina, Desrina; Taslihan, Arief; Ambariyanto, Ambariyanto; Jati, Budhi Kuncoro
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 16, No 2 (2011): Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (424.635 KB) | DOI: 10.14710/ik.ijms.16.2.95-102

Abstract

Vibrio alginolyticus adalah bakteri patogen penyebab penyakit vibriosis pada ikan kerapu budidaya di Indonesia. Vaksin Protein Outer Membran (POM) V. alginolyticus telah terbukti imunogenik pada ikan kerapu. Penelitian ini bertujuan untuk menentukan pengaruh dosis vaksin  terhadap kemampuan POM V. alginolyticus 74 kDa dalam merangsang kerja sistem kekebalan spesifik ikan dan menentukan efek dosis terhadap perlindungan yang dihasilkan. POM(74 kDa) diisolasi dengan metoda sonikasi dan SDS-PAGE, dan dimurnikan dengan metoda elektroelusi. Vaksin diberikan dengan metoda suntik intraperitoneal ke ikan kerapu ukuran 8-10 cm (berat 1013 g) dengan dosis 0 (kontrol), 5, 10 dan 15 µg/0, 1 ml PBS/ 10 g ikan (n= 30 ekor/dosis).  Ikan kontrol disuntik dengan 0,1 ml PBS steril. Satu minggu kemudian ikan disuntik booster dengan cara dan dosis yang sama.  Dua  minggu  setelah  booster  dilakukan  uji  tantang  dengan  dengan  menyuntikkan  bakteri  Vibrio alginolyticus 8 secara intramuskular dengan dosis 0,1 ml X 109 sel/ml, dan ikan dipelihara selama 2 minggu. Jumlah ikan yang mati selama masa uji tantang dihitung untuk menentukan Relative Percentage Survival (RPS). Titer antibodi diukur sebelum percobaan dan setiap minggu selama penelitian. Ke 3 dosis yang diberikan efektif dalam merangsang respon kekebalan humoral ikan kerapu dan menghasilkan kekebalan yang melindungi yang hampir sama yang terlihat dari nilai RPS untuk dosis 5, 10 dan 15 µg masing masing 72, 87 dan 72%. Kata kunci: vaksin POM,  kerapu, dosis  Vibrio alginolyticus is a causative agent of  vibriosis of cultured grouper in Indonesia.  It has been reported  that the Outer Membrane Protein (OMP) of V. alginolyticus vaccine was immunogenic on grouper.  Vaccine dose is important in determining the ability of vaccine to conferred protective immunity. The objectives of the present research was to determine effect of vaccine doses on (1)  the specific immune response of grouper and (2) conferring protective immunity of grouper. OMP V. alginolyticus (74 kDa) was isolated by sonication and SDSPAGE, and purified by mean  electroelution. Vaccine was delivered by intraperitoneal injection to grouper juvenile (8 - 10 cm long  and  weigh10 - 13 g) in three doses;  0 (kontrol), 5, 10 dan 15 µg/0, 1 ml PBS/ fish (n= 30 fish/ dose).  Control fish were injected with 0,1 ml sterile PBS steril. One week later, booster  was given in the same manner as the primary vaccination. Two weeks following booster (week 4), fish were challenge with  V. alginolyticus 8 by intramuscular injection (0,1 ml X 109 sel/ml) and fish were maintained for two weeks. Fish mortality pos challenge test was counted to calcualte the Relative Percentage Survival (RPS). Antibodi titer was measured before vaccination and weekly for 4 week. All three doses tested were effective to  trigger the specific immune response of grouper dan conferred protective immunity with similar degree as shown by the RPS for dose 5, 10 dan 15 µg were 72, 87 dan 72% respectively. Key words: vaccine, OMP, grouper, dose.
TRANSMISSION OF WHITE SPOT SYNDROME VIRUS AND POSSIBLE USE OF PHYSICAL BARRIER AS PREVENTIVE MEASURE (TRANSMISI WHITE SPOT SYNDROME VIRUS DAN PENGGUNAAN BARIER FISIK SEBAGAI UPAYA PENCEGAHAN) Taslihan, Arief; Sumiarto, Bambang; Nitimulyo, Kamiso H
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 18, No 3 (2013): Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (338.074 KB) | DOI: 10.14710/ik.ijms.18.3.119-126

Abstract

Penyakit bercak putih viral hingga saat ini masih menjadi masalah dalam budidaya udang. Munculnya penyakit tersebut diikuti kematian massal, sehingga menimbulkan kerugian besar. Penyakit yang disebabkan white spots syndrome virus (WSSV) menular cepat dari satu petakan tambak ke petakan lain. Penelitian bertujuan melakukan uji kuantitas WSSV pada transmisi virus baik melalui air dan kohabitasi. Metode penelitian adalah bioassay dilakukan skala laboratorium. Penularan melalui air disimulasi pada akuarium disekat dengan 3 jenis kasa berukuran pori berbeda, yaitu 300?, 700? dan 2 mm. Kohabitasi dilakukan dengan memelihara udang terinfeksi WSSV secara buatan dengan udang dan moluska sehat. Hasil penelitian didapatkan bahwa WSSV menimbulkan infeksi pada udang sehat yang ditempatkan terpisah dari udang sakit menggunakan sekat kasa. Virus bercak putih juga menular secara kohabitasi udang sakit dengan udang sehat baik dari udang windu ke udang windu (sejenis) maupun udang windu ke udang vannamei (berlainan jenis). Hasi penelitian menunjukkan bahwa trisipan bukan karier WSSV, karena tidak menularkan. Analisis LT-50 (lethal time 50%) didapatkan bahwa udang yang diuji tantang WSSV melalui inkubasi dengan air mengandung ekstrak WSSV didapatkan konsentrasi 2,75x102 WSSV copy.mL-1 menyebabkan kematian 50% dalam waktu 108 jam atau hampir lima hari. Penggunaan kasa putih meskipun tidak sepenuhnya menahan, dapat menghambat sebagian transmisi WSSV. Hasil kajian memberikan gambaran tentang kecepatan penyebaran WSSV di lingkungan budidaya udang serta memberikan panduan bagaimana mengendalikan WSSV. Kata kunci: penyakit, transmisi WSSV, udang, kohabitasi, trisipan White spot viral disease has devastated shrimp industry in Indonesia. The emergence of this disease is always followed by massive death causing huge losses. Disease is caused by a virus namely White spots syndrome virus (WSSV) is rapidly transmitted from one pond to other ponds. This study aims to quantify WSSV upon transmission process at different route of transmission either through water, the cohabitation ant to cerithidae. A model has developed to fascilitate transmission through water. Aquarium capacity of 60 liter use in this research, each made into two compartment with separation by 3 different screen with pore sizes, 300?, 700? and 2 mm. Healthy tiger shrimp Penaeus monodon, reared in one compartment and artifitially diseased shrimp in another compartment. Cohabitation was done by rearing healthy shrimp of tiger shrimp and vannamei shrimp together with artifially infected shrimp with WSSV. For transmission through snails, snail fed with infected shrimp and reared together with infected live shrimp. Transmission also done through bathing healthy shrimp into water contained WSSV extract. The result showed that WSSV is able to cause infection in healthy shrimp eventhough are spaced apart from diseased shrimp using different mesh size screen. White spot virus can also be transmitted by cohabitation from diseased tiger shrimp either to tiger shrimp (same species) or to vannamei shrimp (different species). The result showed that snail is not a career for WSSV. LT-50 of challenge of shrimp with WSSV through incubation with water contained of WSSV extract found that innocula at concentration 2,75 x 105 WSSV copy.?l-1 causing mortality at 50% within 108 hours. White screen, eventhough not fully efective, but still can retarded WSSV transmission. Result of study provide a greater understanding of how the virus be transmitted in the shrimp farm, and as guidance strategy in controlling in shrimp aquaculture. Keywords: disease, transmission of WSSV, cohabitation, shrimp, snail
CLUSTER MANAGEMENT TO PREVENT TRANSMISION OF WHITE SPOT SYNDROME VIRUS IN EXTENSIVE GIANT TIGER SHRIMP FARMING (MANAJEMEN KLASTER TAMBAK UDANG WINDU UNTUK MENCEGAH PENULARAN VIRUS WHITE SPOT SYNDROME) Taslihan, Arief; Sumiarto, Bambang; Nitimulyo, Kamiso H
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 18, No 4 (2013): Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (289.418 KB) | DOI: 10.14710/ik.ijms.18.4.231-238

Abstract

Telah dilakukan kajian terhadap efektifitas managemen model klaster dengan tambak non-udang sebagai tambak penyanggah untuk mencegah transmisi penyakit bercak putih viral (WSSV) pada budidaya udang windu skala tradisional. Kajian dilakukan di tambak udang tradisional di wilayah kabupaten Demak, provinsi Jawa Tengah. Penelitian dilakukan pada lima petak tambak perlakuan dan tujuh petak tambak sebagai kontrol. Tambak udang pada kelompok perlakuan menerapkan sistem klaster, yaitu tambak dikelilingi dengan petak berisi ikan sebagai komponen biosekuriti untuk mencegah penularan WSSV dari tambak sekitarnya. Tambak kontrol tidak menggunakan petak non-udang sebagai komponen biosekuriti, dan dikelola dengan teknologi yang biasa dilakukan oleh pembudidaya setempat. Hasil penelitian menunjukkan bahwa tambak yang dikelilingi dengan tambak non-udang dapat dipanen dalam waktu pemeliharaan 105,6±4,5 hari lebih lama secara nyata dibanding tambak kontrol yang dipanen pada hari ke 60,9±16,0 karena wabah penyakit, sintasan (survival rate) yang diperoleh adalah 77,6±3,6 %, lebih besar secara nyata dibandingkan kontrol yang hanya 22,6±15,8 %, serta produksi udang 425,1±146,6 kg.ha-1 jauh lebih tinggi dibandingkan kontrol yang hanya 54,5±47,6 kg.ha-1. Pada kajian tersebut tambak non-udang ditebari dengan tilapia dan kakap. Hasil penelitian menunjukkan bahwa melalui penerapan Better Management Practices (BMP) dengan cara tambak udang dalam model klaster yang dikelilingi dengan tambak non-udang terbukti efektif mencegah penularan WSSV dari tambak tradisional sekitarnya. Kata kunci: budidaya udang tradisional, windu, biosekuriti, manajemen klaster  White spot syndrome virus (WSSV) has become epidemic in Indonesia and affecting shrimp production lost in shrimp farm. Virus has transmitted from one to other ponds, mostly by crustacean, but more often transmit through water from affected pond. A cluster model, consist of two and three ponds surrounded by non-shrimp growing pond as biosecurity has developed. The model aim to prevent white spot virus transmission in giant shrimp extensive pond. The study was conducted in two sites at Demak district, Central Java province. Cluster consist of three shrimp ponds in site I, and cluster consist of two shrimp ponds, each surrounded by non-shrimp growing ponds. As control we also compare to 5 extensive shrimp ponds in site I and other three shrimp grow out ponds in site II, with neither no cluster system nor surrounded by non-shrimp pond as biosecurity. Result of the study shown that cluster of shrimp ponds surrounded  by non-shrimp pond harvested at DOC 105,6±4,5 days significantly longer than that of control that harvested at 60,9±16,0 days because of outbreak, survival rate at 77,6±3,6 %, significantly higher than that of control at 22,6±15,8 % and shrimp production of 425,1±146,6 kg.ha-1 significantly higher than that of control at 54,5±47,6 kg.ha-1. These results suggest that implementation of Better Management Practices (BMP) by arranging shrimp ponds in cluster and surrounding by non-shrimp ponds proven effectively prevent WSSV transmission from traditional shrimp ponds in surrounding area. Keywords: extensive shrimp pond, giant tiger prawn, biosecurity, cluster management
PROFIL HETEROGENITAS GENETIK INDUK UDANG WINDU (Penaeus monodon) TURUNAN F1 MELALUI ANALISIS DNA MITOKONDRIA-RFLP DAN RAPD Prastowo, Bambang W.; Rahardianti, Rahayu; Nur, Evi Maftuti; Taslihan, Arief
Jurnal Perikanan Universitas Gadjah Mada Vol 11, No 1 (2009)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (159.781 KB) | DOI: 10.22146/jfs.2968

Abstract

Production of high quality of shrimp broodstock needs proper domestication and improvement of their genetic quality. MCBAD Jepara has conducted research to evaluate genetic heterogeneity of Penaeus monodon broodstock F1 using Restriction Fragment Length Polymorphism (RFLP) and Random Amplified Polymorphic DNA (RAPD) analysis. For RFLP analysis, amplification product of 16SrDNA of mitochondrial DNA was digested with restriction enzymes. According to the RFLP analysis, heterogeneity value of P. monodon F1 broodtock population was 0.0422; male F1 population was 0.0613 and female F1 population was 0.1252. The primer used in RAPD analysis was OPA 2. According to the RAPD analysis, heterogeneity value of P. monodon F1 broodstock population was 0.0417; male F1 population was 0.0653 and female F1 population was 0.1104. The results from these researches have shown that either RFLP or RAPD can be used as a family specific marker for Penaeus monodon.