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DUPLEX POLYMERASE CHAIN REACTION UNTUK DETEKSI SIMULTAN KOI HERPESVIRUS DAN AEROMONAS HYDROPHILA PADA IKAN MAS (CYPRINUS CARPIO) Novita, Hessy; Sugiani, Desy; Taukhid, Taukhid; Sumiati, Tuti
Jurnal Riset Akuakultur Vol 15, No 1 (2020): Maret, 2020)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/jra.15.1.2020.%p

Abstract

Koi herpesvirus (KHV) dan Aeromonas hydrophila adalah patogen yang dapat mengkoinfeksi ikan mas secara bersamaan. Penelitian ini bertujuan untuk mengembangkan metode duplex polymerase chain reaction (dPCR), deteksi simultan untuk diagnosis KHV dan bakteri Aeromonas hydrophila pada ikan mas. Dua pasang primer yang menargetkan sekuen spesifik SphI dan gen aerolisin, yang sering digunakan untuk mendeteksi KHV dan A. hydrophila dalam uji reaksi tunggal PCR dan menghasilkan target pita PCR 290 bp dan 417 bp. Hasil penelitian menunjukkan bahwa metode duplex PCR dapat mendeteksi ganda infeksi KHV dan A. hydrophila pada ikan mas dan metode ini lebih efektif mendeteksi dua patogen secara bersamaan dalam satu reaksi PCR pada suhu pradenaturasi, 94°C selama dua menit, denaturasi pada suhu 95°C selama satu menit, annealing pada suhu, 55°C selama satu menit, dan 72°C selama satu menit, dengan 30 siklus amplifikasi dan final extention pada suhu 72°C selama lima menit. Metode dPCR untuk deteksi simultan kedua patogen adalah salah satu metode yang dapat diaplikasikan untuk deteksi koinfeksi virus dan bakteri dalam satu reaksi PCR.Koi herpesvirus (KHV) and Aeromonas hydrophila are pathogens that can co-infect common carp. This study aimed to develop a duplex polymerase chain reaction (dPCR) method to detect KHV and Aeromonas hydrophila in common carp simultaneously. Two pairs of primers targeted the specific sequences of SphI and aerolysin genes, often used in detecting KHV and A. hydrophila, in a single PCR reaction test and produced target bands of PCR 290 bp and 417 bp. This proposed method was more effective in simultaneously detecting the two pathogens in one PCR reaction at pre-naturation temperature of 94°C for two minutes, denaturation at 95°C for one minute, annealing at temperature, 55°C for one minute, and 72°C for one minute, with 30 cycles of amplification and final extension at 72°C for five minutes. The findings showed that the duplex PCR method could be used to double detect KHV and A. hydrophila infection in common carp. The duplex PCR method for simultaneous detection of both pathogens is one method that can be applied for the detection of co-infection of viruses and bacteria in a PCR reaction.
CERITA RAKYAT PULAU MAYA KABUPATEN KAYONG UTARA (KAJIAN STRUKTUR, FUNGSI, KONTEKS, DAN NILAI BUDAYA) Sumiati, Tuti; Effendy, Chairil; Martono, Martono
Jurnal Pendidikan dan Pembelajaran Khatulistiwa Vol 8, No 9 (2019): September 2019
Publisher : Jurnal Pendidikan dan Pembelajaran Khatulistiwa

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (566.232 KB)

Abstract

AbstractThis thesis presents a study that focuses on the study of oral literature, namely folklore. The rapid development of Science and Technology has led to a reduction in speakers and listeners of folklore, for that careful study of folklore texts needs to be done. The focus of the problem in this study is the structure, function, context and cultural value of the story and its implementation plan in learning folklore in schools. The method used is descriptive analysis. This research uses structural approach and sociology of literature. The data source in this study is in the form of folklore told by speakers. The steps of data collection are done by recording, transcribing, editing, translating, identifying and classifying. The data is then interpreted and analyzed by providing interpretations of meaning according to the problem under study. The results of the analysis of this study are: (1) the structure of the story, there is a forward plot, there is a human character, and there is a setting in the form of place, time setting and social setting; (2) the function of the story, found five functions, namely: as a means of entertainment; as a means of education and inculcation of moral values; as a cultural validator; as a means to pass down traditional ways of life, customs and habits; and as a tool so that the norms of society will always be obeyed by their collective members; (3) the narrative context consists of three contexts namely situation context, cultural context, and social context; (4) cultural values consist of human relations with God, human relations with oneself, human relations with the natural surroundings, and human relationships with each other; and (5) the results of this study can be implemented in Bahasa Indoneisa learning in schools because there are positive values that can be learned by students, namely in KD 3.7 identifying the values and content contained in folklore both oral and written, and KD 4.7 retell the contents of folklore heard and read. Keywords: Context, Cultural Values, Folklore, Function, Structure
APLIKASI VAKSIN Streptococcus agalactiae UNTUK PENCEGAHAN PENYAKIT STREPTOCOCCOSIS PADA BUDIDAYA IKAN NILA (Oreochromis niloticus) [Application of Streptococcus agalactiae vaccine to prevent streptococcosis on tilapia culture, Oreochromis niloticus] Taukhid, Taukhid; Lusiastuti, Angela Mariana; Sumiati, Tuti
BERITA BIOLOGI Vol 13, No 3 (2014)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (110.857 KB) | DOI: 10.14203/beritabiologi.v13i3.662

Abstract

The research with the aim to know the effectivity (yield gap) of the application of Streptococcus agalactiae vaccine (pure whole cell) in prevention of streptococcosis on tilapia (Oreochromis niloticus) culture has been carried out. The isolate of S.agalactiae – N14G was used as a master seed on vaccine production. Priming vaccination was administered by immersion method, and booster vaccination was taken th place two months latter by oral method. Challenge test at the lethal dose (LD50) against active bacteria was done at 14 days post booster vaccination, and observation was taken place for 14 days post artificial infection. The results of the research showed that the highest survival rate and relative percent survival (RPS) was found in group treated with Streptovac vaccine (S. agalactiae and A. hydrophila combination) (65.58% and 35.36%) followed by S. agalactiae vaccine (52.08% and 10.01%). The lowest survival rate was found in control group (46.75%). The result of confirmation effectivity of the vaccines by challenge test in the laboratory showed that the highest survival rate and relative percent survival (RPS) was found in S. agalactiae vaccine (50.00% dan 37.50%) followed by Streptovac vaccine (40.00% and 25.00%), and the lowest survival rate was found in control group (20.00%). Vaccination is better than the non vaccinated.
Specific Immune Response Kinetics and Mortality Patterns of Tilapia Oreochromis niloticus on Post-Cocktail Vaccination Period against the Infection of Aeromonas hydrophila and Streptococcus agalactiae Sukenda, Sukenda; Sumiati, Tuti; Nuryati, Sri; Lusiastuti, Angela Mariana; Hidayatullah, Dendi
Journal Omni-Akuatika Vol 13, No 2 (2017): Omni-Akuatika November
Publisher : Fisheries and Marine Science Faculty - Jenderal Soedirman University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (214.583 KB) | DOI: 10.20884/1.oa.2017.13.2.279

Abstract

ABSTRACTFish vaccination aims to induce a specific immune response indicated by an increase of antibodies in vaccinated fish. However, in accordance with time the presence of antibodies will continue to decline. The purpose of this study was to determine the kinetics of specifik immune response and trend mortality against Aeromonas hydrophila and Streptococcus agalactiae on tilapia following vaccination with cocktail vaccine. Fish vaccinated through immersion for 30 minutes in a solution of diluted vaccine. Challenge test was performed for three periods, on day 22, 50, and 78 post-vaccination, fish were challenged with single infection of A. hydrophila 108 cfu. mL-1 and S. agalactiae 104 cfu. mL-1 and co-infection of both bacteria by intraperitoneal. During rearing, the blood fish were taken for determining of serum antibodies, and its  measured by ELISA. The results showed that the concentration of specific antibodies vaccinated fish were significantly higher than the control. The basal antibody levels of A. hydrophila before vaccination were higher than S. agalactiae with OD of 0.104 and 0.069 respectively. The maximum  antibody  response  was  reached  within  70  days  of  the  A. hydrophila OD= 0.264 and 56 days against S. agalactiae OD= 0.188. The mortality rate in the control group was significantly higher than vaccinated on all types and each challenge test period. The trend of mortality due to a single infection of A. hydrophila and co-infections occur more quickly than by S. agalactiae. Lowest mortality occurred in the vaccinated group at 50 day tested challenge.Keywords: kinetics antibody, Aeromnas hydrophila, Streptococcus agalactiae, Oreochromis niloticus
SUSCEPTIBILITY OF CYPRINID AND NON-CYPRINID FISH CELL LINES TO KOI HERPESVIRUS (KHV) Sumiati, Tuti; Sunarto, Agus
Indonesian Aquaculture Journal Vol 4, No 2 (2009): (December 2009)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (562.776 KB) | DOI: 10.15578/iaj.4.2.2009.131-137

Abstract

Koi herpesvirus (KHV) is an emerging virus that infects koi and common carp (Cyprinus carpio) with mortality up to 95% within 7 days. The disease is rapidly spreading worldwide including to Indonesia. However, it has only been documented in koi and common carp. The aim of this research was to evaluate the susceptibility of fish cell cultures originated from cyprinid and non-cyprinid fish to KHV. Koi Fin (KF-1) and Koi Tail (KT-2) cell lines derived from koi carp and SSN-1 cells originated from fry of striped snakehead were used in this study. The cells were inoculated with tissue extract of KHV-infected koi carp (experiment 1) and virus stock of KHV (experiment 2). The cultures were incubated at 22oC and the onset and type of cytophatic effect (CPE) were observed for 21 days post inoculation. The results of experiment 1 showed that CPE was observed in KT-2 at day 6 post inoculation. In the experiment 2, however CPE was observed in KF-1 and KT-2 cells at day 4 post infection. CPE was not observed in SSN-1 of either experiment 1 or experiment 2. CPE was characterized by extensive vacuolization of the infected cells. Polymerase chain reaction (PCR) assay of cell and tissue culture supernatants confirmed that KF-1 and KT-2 showing CPE were indeed infected with KHV. The results indicated that KF-1 and KT-2 cells were susceptible and SSN-1 was resistant to KHV. The implication of these findings was also discussed in the paper.
THE EFFECTS OF STORAGE TEMPERATURE AND TYPES OF MEDIA ON THE SURVIVAL RATE OF S. agalactiae BACTERIOPHAGE Lusiastuti, Angela Mariana; Sumiati, Tuti; Priadi, Adin
Indonesian Aquaculture Journal Vol 3, No 2 (2008): (December 2008)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (96.544 KB) | DOI: 10.15578/iaj.3.2.2008.147-149

Abstract

The successful effect of phages in phage therapy has been proven by an increase of the presence of phages in survivors. Seven previously isolated phages were used to examine the effects of storage temperature and types of media on their survival rate. Phages PSaT-18, PSaT-19, PSaT-20 and PSaT-21 isolated from a Streptococcosis-infected Tilapia (Oreochromis niloticus) were used in this study. Phages PSaG-1, PSaG-2 and PSaG-3 were isolated from a diseased Gouramy (Osphronemus gouramy). All of the phages were stored in refrigerator at temperature of 8oC and kept in two types of media, Brain Heart Infusion Broth (BHIB) and BHIB enriched with glycerol. To determine the lytic activity of the phages, 20 μL of each phage suspension was poured onto BHIA plate and incubated at 25oC for 24 to 48 hours. The result showed that phages PSaT-18, PSaT-19, PSaT-20 and PSaT-21 have lytic activity against S. agalactiae and S. iniae than phages PSaG-1, PSaG-2, and PSaG-3. Phages PSaT-18, PSaT-19, PSaT-20, and PSaT-21 always survived up to three weeks and phage PSaT-18 in BHIB have the highest potential as a candidate of phage therapy for Streptococcosis.
EFIKASI VAKSIN IN-AKTIF BAKTERI Aeromonas hydrophila-AHL0905-2 (HYDROVAC) dan Streptococcus agalactiae-N14G (STREPTOVAC) UNTUK PENCEGAHAN PENYAKIT BAKTERIAL PADA IKAN BUDIDAYA AIR TAWAR Taukhid, Taukhid; Purwaningsih, Uni; Sugiani, Desy; Sumiati, Tuti; Lusiastuti, Angela Mariana
Jurnal Riset Akuakultur Vol 10, No 4 (2015): (Desember 2015)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (112.754 KB) | DOI: 10.15578/jra.10.4.2015.541-551

Abstract

Penelitian bertujuan untuk mengetahui efektivitas penggunaan vaksin hydrovac dan streptovac untuk pencegahan penyakit bakterial, motile aeromonad septicaemia (MAS) dan streptococcosis pada beberapa jenis ikan budidaya air tawar. Pengujian dilakukan pada skala laboratorium dan lapang. Jenis ikan uji yang digunakan adalah ikan lele, nila, dan gurami. Vaksinasi ikan dilakukan melalui teknik perendaman dengan dosis dan periode sesuai instruksi penggunaan yang tertera pada label produk kedua jenis vaksin tersebut. Efektivitas vaksin dievaluasi berdasarkan pendekatan nilai persen sintasan dan selanjutnya dihitung nilai relative percentage survival (RPS). Hasil penelitian diketahui bahwa nilai RPS vaksin hydrovac pada skala laboratorium pada ikan lele, nila, dan gurami masing-masing sebesar 85,45%; 65,78%; dan 52,28%. Nilai RPS yang dicapai oleh vaksin streptovac terhadap ikan nila sebesar 54,53%. Sementara, nilai RPS vaksin hydrovac pada skala lapang untuk jenis ikan lele, nila, dan gurami masing-masing 70,15%; 52,43%; dan 42,43%; sedangkan nilai RPS yang dicapai oleh vaksin streptovac adalah 40,41%.
ISOLASI KOI HERPESVIRUS (KHV) DARI BEBERAPA ORGAN TARGET DENGAN MENGGUNAKAN KULTUR SEL KT-2 Sumiati, Tuti; Sunarto, Agus
Jurnal Riset Akuakultur Vol 7, No 1 (2012): (April 2012)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1208.144 KB) | DOI: 10.15578/jra.7.1.2012.93-100

Abstract

Kasus kematian massal pada ikan mas dan koi (Cyprinus carpio) yang disebabkan oleh koi herpesvirus (KHV) terjadi sejak tahun 2002 dan masih berlangsung hingga sekarang. Pemilihan sampel yang tepat sangat penting untuk mendeteksi dan mengidentifikasi penyakit KHV tersebut. Tujuan dari penelitian ini adalah untuk mengetahui jaringan yang menjadi target infeksi KHV dengan cara isolasi virus menggunakan kultur sel KT-2. Kultur sel diinokulasi dengan ekstrak jaringan organ target (otak, mata, insang, ginjal, limfa, hati, jantung, dan usus, serta gabungan insang, ginjal, dan limfa) dan diinkubasi pada suhu 25oC selama 14 hari. Kerusakan sel terjadi pada kultur sel yang diinokulasi dengan ekstrak dari jaringan insang, ginjal dan gabungan organ insang, ginjal, dan limfa. Uji PCR dari media kultur dan sel yang mengalami CPE menunjukkan bahwa CPE disebabkan oleh KHV.
ISOLASI BAKTERIOFAGA ANTI Streptococcus agalactiae DARI IKAN NILA (Oreochromis niloticus) Lusiastuti, Angela Mariana; Purwaningsih, Uni; Sumiati, Tuti
Jurnal Riset Akuakultur Vol 5, No 2 (2010): (Agustus 2010)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (201.907 KB) | DOI: 10.15578/jra.5.2.2010.237-243

Abstract

Infeksi Streptococcus merupakan salah satu penyakit serius pada ikan yang disebabkan oleh bakteri gram positif. Infeksi oleh streptococcus beta-hemolitik paling sering dilaporkan menginfeksi ikan. Di antara streptococci beta-hemolitik, Streptococcus iniae penyebab septicemia, meningoencefalitis, dan kematian pada ikan budidaya. Selain itu, Streptococcus agalactiae juga menyebabkan streptococcosis parah pada ikan nila. Alternatif yang bisa digunakan untuk terapi infeksi streptococcosis adalah dengan penggunaan bakteriofaga yang merupakan virus yang hidup pada bakteri. Tujuan penelitian ini adalah isolasi bakteriofaga S. agalactiae sebagai kandidat agen terapi yang memberikan efek protektif melawan infeksi streptococcosis. Faga diisolasi dari Brain Heart Infusion Agar (BHIA) yang sudah ditanami dengan 15 isolat S. iniae dan S. agalactiae. Isolat S. iniae dan S. agalactiae diisolasi dari ikan sakit dengan gejala klinis Streptococcosis. Setelah itu diidentifikasi dengan pewarnaan Gram, tes katalase, pertumbuhan pada agar darah dan API 20 Strep System. Pertumbuhan faga ditunjukkan dengan adanya zona lisis pada tempat yang ditetesi dengan sampel cairan usus dari ikan nila sehat. Faga yang tumbuh lalu dikoleksi secara steril, disentrifus dan supernatannya difiltrasi dengan membran filter 0,45 µm dan disimpan pada suhu 4oC. Dari 15 isolat S. iniae dan S. agalactiae hanya satu isolat yaitu PSaT-18 yang menunjukkan zona lisis seperti yang ditunjukkan pada cawan petri isolat kontrol S. iniae. Zona lisis tersebut timbul akibat adanya faga yang memberikan proteksi terhadap S. iniae dan S. agalactiae. Zona lisis yang tidak jernih disebabkan konsentrasi faga yang terlalu rendah akibat dilakukan pengenceran pada proses filtrasi. Faga yang diperoleh selanjutnya akan dilakukan uji in vitro dan in vivo untuk mengetahui efektivitasnya.Streptococcal infection is a serious disease in fish caused by gram positive bacteria. The causative agent is Streptococcus b-hemolytic. Streptococcus iniae, a b-hemolytic bacterium is the main causative agent of septicemia, meningoencephalitis, and fish mortality. Moreover, the other causative agent is S. agalactiae. Bacteriophages which are viruses that live on bacteria can be selected and used as therapy for Streptococcosis. The aim of this research is to isolate bacteriophage of S. agalactiae as therapeutic agent candidate giving protecting effect for Streptococcosis. Phages was isolated from Brain Heart Infusion Agar (BHIA) were obtained from cultures of S. iniae and S. agalactiae. Strains S. iniae and S. agalactiae were isolated from Streptococcosis fish. The predominant types of colonies were subcultured and subjected to biochemical and physiological tests such as Gram staining, catalase test, hemolytic activity in blood agar and API 20 Strep System. Phages were isolated by using a double agar layer method. A zona lysis with plaques was removed from the plate, centrifuged and the supernatant was filtered through a 0.45 µm membrane filter and stored at 4oC. From 15 isolates of S. iniae and S. agalactiae, only one isolate PSaT-18 showed lysis zone. The lysis zone developed because the phages give protection to S. agalactiae. The lysis zone is not clear because the phage concentration is too low and the dilution in filtration process is too high. Later, these phages were used for invitro and invivo test.
SEDIAAN VAKSIN Mycobacterium fortuitum ISOLAT LOKAL YANG EFEKTIF UNTUK PENCEGAHAN PENYAKIT Mycobacteriosis PADA IKAN GURAMI, Osphronemus gouramy Purwaningsih, Uni; Taukhid, Taukhid; Lusiastuti, Angela Mariana; Sugiani, Desy; Sumiati, Tuti
Jurnal Riset Akuakultur Vol 10, No 3 (2015): (September 2015)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (353.914 KB) | DOI: 10.15578/jra.10.3.2015.423-433

Abstract

Mycobacteriosis merupakan penyakit yang bersifat kronis progresif yang rentan menyerang ikan gurami, dengan tingkat prevalensi mencapai 30%-80%. Berbagai upaya telah dilakukan untuk mencegah dan mengendalikan penyakit tersebut dengan menggunakan antibiotik, bahan kimia maupun terapi herbal namun belum memberikan hasil yang optimal. Vaksinasi diharapkan mampu menjadi solusi alternatif dan aplikatif untuk pencegahan penyakit Mycobacteriosis pada ikan gurami. Penelitian ini bertujuan untuk memperoleh jenis sediaan vaksin M. fortuitum yang tepat untuk mencegah penyakit Mycobacteriosis pada ikan gurami. Isolat M. fortuitum kode 31 digunakan sebagai isolat kandidat vaksin. Inaktifasi vaksin dilakukan dengan sonikasi dan neutral buffer formalin 3%. SDS PAGE terhadap sediaan sel utuh dan broth menunjukkan jumlah pita protein yang lebih variatif. Berdasarkan uji innocuity dan uji sterility terhadap berbagai sediaan vaksin menunjukkan bahwa vaksin terbukti aman dan tidak menyebabkan efek samping pada ikan gurami. Peningkatan titer antibodi terjadi 14 hari pasca vaksinasi. Titer antibodi pada perlakuan vaksin menunjukkan perbedaan yang nyata (P<0,05) dibanding kontrol pasca uji tantang. Kematian ikan pasca uji tantang dengan menggunakan bakteri M. fortuitum 106 cfu/mL menunjukkan pola kematian yang bersifat kronis. Kematian mulai terjadi setelah hari ke-19 pasca uji tantang. Ikan gurami yang divaksinasi dengan vaksin sel utuh M. fortuitum menunjukkan hasil terbaik dengan tingkat sintasan sebesar 83,33% dan relative percent survival (RPS) sebesar 66,67%.