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Skrining Bakteri yang Berasosiasi dengan Spons Jaspis sp. Sebagai Penghasil Senyawa Antimikroba Abubakar, Hermawaty; Wahyudi, Aris Tri; Yuhana, Munti
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 16, No 1 (2011): Jurnal Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

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Abstract

Organisme bentik laut seperti spons, seringkali hidup berasosiasi dengan bakteri yang menghasilkan senyawa antimikroba.  Penelitian  ini  bertujuan  untuk  mengetahui  kemapuan  antagonis  isolat-isolat  bakteri  yang berasosiasi dengan spons Jaspis sp. terhadap beberapa bakteri patogen, dengan metode skrining secara kualitatif. Sebanyak 32 (45,71%) dan 20 (29,41%) isolat yang berasal dari bagian mesohyl dan permukaan Jaspis sp. menunjukkan kemampuan antimikroba, karena mampu menghambat pertumbuhan Staphylococcus aureus, Vibrio harveyii, Escherichia coli, Pseudomonas aerogenosa, EPEC K-11, Candida albicans, and C. tropicalis. Uji fenotipik dilakukan pada beberapa isolat dengan aktivitas antimikroba terbaik, yaitu SAB E-8, SAB E-33, SAB E-35, SAB E-38, SAB E-40 dan SAB S-43. Hasil pewarnaan Gram menujukkan isolat  SAB E-8, SAB E35, and SAB E-40 adalah Gram negatif, sedangkan isolat SAB E-33, SAB E-38, and SAB S-43 adalah gram positif yang dilanjutkan dengan identifikasi parsial (pengecatan gram dan uji katalase) untuk kelompok Bacillus.Kata kunci: Bakteri, Asosiasi, Jaspis sp., antimikroba Living benthic marine organisms such as sponges are frequently assosiated with as bacteria that may be produce antimicrobial compounds. This study aims to determine antagonistic of bacterial isolates that associated sponge Jaspis sp., with a qualitative screening method. Screening of bacteria from marine sponge Jaspis sp. which have bility to produce antibacterial subtances was investigated. There are 32 (45,71%) and 20 (29,41%) isolates from mesohyl and surface sponge respectively. Those isolated bacterial showed the antibacterial activity against Staphylococcus aureus, Vibrio harveyii, Escherichia coli, Pseudomonas aerogenosa, EPEC K-11, Candida albicans, and C. tropicalis. However, use of a few additional simple phenotypic tests for those isolate can be used to differentiate among isolates. The simple phenotypic test divided two ways based on staining gram. Gram negative bacteria were desingned SAB E-8, SAB E-35, and SAB E-40 and gram positive bacteria were desingned SAB E-33, SAB E-38, and SAB S-43. Parsial identification that directed to Bacillus was used for positive gram bacteria, involve gram staining, endospora staining and katalase test. Key words: Bacteria, Assosiation, Jaspis sp, antimicrobe
Biocontrol agents in aquaculture: Production and their application Yuhana, Munti
Jurnal Akuakultur Indonesia Vol 9, No 1 (2010): Jurnal Akuakultur Indonesia
Publisher : Jurnal Akuakultur Indonesia

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Abstract

The use of biocontrol agents have become commonly accepted as a ´natural weapon´ in aquaculture, either as an ecological bioremediator for low environmental quality, as well as the health promoter for cultured organisms. Biocontrol agents, which consist of beneficial microorganisms, are commonly applied as biosupplements in feeds. They are potential in replacing the use of antibiotic in inhibiting the pathogenic organisms. Therefore, their application has revealed the potential as an effective strategy to reduce the over use of antibiotics in controlling the pathogenic agents, avoid the spreading the drug resistance, or environmental deterioration of the negative effect by killing useful microorganisms. The development of suitable technology for microbial agents production, viability and stability, is a key area of research for industrial production. Production of biocontrol agents should be based on the microbial criteria, and the ability to withstand stress during processing and storage of products is crucial. This review makes an overview of biocontrol agents selection studies including techniques for isolation/identification, selection, production and its application in order to be accepted as a valuable product in aquaculture. Key words: Biocontrol agents, beneficial microorganisms, aquaculture.   ABSTRAK Pemanfaatan agen biokontrol telah dapat diterima secara luas di dunia akuakultur sebagai senjata alami, baik sebagai bioremediator ekologis untuk memperbaiki kualitas lingkungan yang rendah maupun sebagai promoter/pendukung kesehatan untuk organisme-organisme yang dibudidayakan. Agen-agen biokontrol yang terdiri atas berbagai mikroorganisme yang menguntungkan umumnya diaplikasikan sebagai biosuplemen dalam pakan. Mikroorganisme tersebut potensial dalam menggantikan penggunaan antibiotik dalam proses penghambatan terhadap organisme patogenik. Oleh karena itu, aplikasinya yang sangat potensial dapat berguna sebagai strategi efektif untuk mereduksi penggunaan berlebih dari antibiotik dalam pengendalian agen patogenik, mencegah penyebaran sifat resistensi terhadap obat-obatan, atau mencegah kerusakan lingkungan dari kematian mikroorganisme yang bermanfaat. Pengembangan teknologi yang sesuai untuk produksi, viabilitas dan stabilitas dari agen biokontrol tersebut, merupakan kunci dari penelitian untuk produksi masal pada skala industri. Proses produksi dari agen biokontrol harus berdasar pada kriteria mikrobiologis. Sifat-sifat ketahanannya terhadap stress selama pemrosesan maupun penyimpanan produk adalah penting. Pada makalah ini disajikan teknik-teknik untuk isolasi, identifikasi, produksi dan aplikasi agen biokontrol untuk dapat diterima sebagai produk yang bernilai ekonomis dalam akuakultur. Kata kunci: Agen biokontrol, mikroorganisme yang menguntungkan, akuakultur.
Existing of White Spot Syndrome Virus (WSSV), Taura Syndrome Virus (TSV) and Infectious Hypodermal Haematopoitic Necrosis Virus (IHHNV) in White Shrimp Litopenaeus vannamei Reared at Intensive Tambak System in Bakauheni, Lampung Selatan Dwinanti, S.H.; Yuhana, Munti
Jurnal Akuakultur Indonesia Vol 8, No 2 (2009): Jurnal Akuakultur Indonesia
Publisher : Jurnal Akuakultur Indonesia

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Abstract

Development of white shrimp, Litopenaeus vannamei culture to increase production yield should take attention in prevention of viral outbreak which  is a main factor caused harvesting failure.  Viruses that usually infect shrimp are white spot syndrome virus (WSSV), taura syndrome virus (TSV) dan infectious hypodermal hematopoietic necrosis virus (IHHNV).  To prevent virus infection and reduce risk of fail to harvest, an early warning and observation system of availability of pathogen should be taken in order to know the shrimp condition during culture process.  This study was performed to examine existing of WSSV, TSV and IHHNV in shrimp reared in tambak by PCR and histopathology methods. Samples were taken every two weeks from tambak culture at Bakauheni, Penengahan, Lampung Selatan.  Generally, WSSV, TSV, and IHHNV were found in shrimp from tambak and water around tambak.  Virus infection was first detected in shrimp derived from tambak on 66 days after crop and still exists until 107 days after crop.  Hepatopancreatic cell disorder  caused by the three viruses were found in cell nuclei that became bigger, necrosis in cytoplasm and inclusion body at nuclei.  The existing of virus in outside of tambak indicated that virus was horizontally transmitted. Keywords: WSSV, TSV, IHHNV, white shrimp   ABSTRAK Perkembangan budidaya udang vaname Litopenaeus vannamei dalam usaha meningkatkan hasil produksi harus memperhatikan keberadaan penyakit viral yang menjadi penyebab utama kegagalannya. Virus yang biasa menyerang vaname antara lain white spot syndrome virus (WSSV), taura syndrome virus (TSV) dan infectious hypodermal hematopoietic necrosis virus (IHHNV). Untuk mengantisipasi penyebaran virus dan mengurangi resiko kegagalan produksi diperlukan usaha pencegahan dengan melakukan peringatan dini (early warning) dan pemantauan tambak terhadap keberadaan patogen tersebut selama masa budidaya. Penelitian ini dilakukan untuk mengetahui keberadaan WSSV, TSV dan IHHNV yang menginfeksi udang vaname dengan analisa polymerase chain reaction (PCR) dan histopatologis. Pengambilan sampel dilakukan sebanyak delapan kali setiap dua minggu di tambak intensif udang vaname Litopenaeus vannamei di Bakauheni, Penengahan, Lampung Selatan. Secara umum, WSSV, TSV dan IHHNV telah ditemukan pada tambak serta di perairan sekitar lokasi tersebut. Infeksi virus pertama kali terdeteksi di tambak ketika udang berumur 66 hari setelah penebaran dan terdeteksi pula pada waktu pengamatan selanjutnya sampai udang berumur 107 hari setelah penebaran. Kerusakan akibat serangan ketiga jenis virus tersebut terjadi pada bagian inti sel yang mengalami pembesaran, nekrosis pada sitoplasma dan badan inklusi yang menekan inti sel. Pola transmisi virus yang terjadi adalah secara horizontal yang didukung oleh terdeteksinya virus tersebut pada udang di luar tambak.   Kata kunci : WSSV, TSV, IHHNV, udang vaname
Free-Living Ice-Nucleating Active Bacteria from High Mountain Lake Habitats YUHANA, MUNTI; HANSELMANN, KURT
Microbiology Indonesia Vol 2, No 3 (2008): December 2008
Publisher : Indonesian Society for microbiology

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Abstract

We collected the culturable heterotrophic bacteria from oligotrophic high mountain lake habitats and tested their capability to induce ice formation. Direct plating was carried out using low-nutrient medium at a temperature of between 3 and 4°C. As many as 84 isolates were recovered from glacial ice and natural biofilm growing on granite rocks surface. Six out of 84 isolates were capable of expressing the ice-nucleation phenotype. After autoclaving the cell suspension at 121°C for 15 min, isolate J78 was still able to retain the ability for ice formation. Heat-stable ice nuclei produced by ice-nucleating active bacteria have potential applications in biotechnology. Characterization of INA bacteria was performed employing live-dead Gram staining and molecular methods. Universal primers for Bacteria (S-D-Bact-0008-b-S-20 and S-D-Bact-1524-a-A-18) were used for PCR to amplify almost the full length of the 16S rRNA genes of selected INA isolates. Restriction fragment length polymorphism analysis resulted in 2 unique patterns, as represented by J43 and J83, respectively. Based on DNA sequencing of 16S rRNA gene, isolate J43 (GeneBank accession no. AJ864852) was closely related to Pseudomonas mephitica (99.2% sequence similarity) and Janthinobacterium lividum (99% similarity), whereas isolate J83 (GeneBank accession no. AJ864859) showed 100% sequence identity to Pseudomonas fluorescens.
Efficacy of DNA vaccine encoding koi herpesvirus glycoprotein GP-25in common carp juvenile by immersion Nuswantoro, Soko; Alimuddin, .; Yuhana, Munti; Santika, Ayi; Nuryati, Sri; Zainun, Zakki; Mawardi, Mira
Jurnal Akuakultur Indonesia Vol 11, No 1 (2012): Jurnal Akuakultur Indonesia
Publisher : Jurnal Akuakultur Indonesia

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Abstract

Koi herpesvirus (KHV) is a herpesvirus that particularly infects and causes mass mortality to koi and common carp. Therefore, the protection of common carp from KHV infection is urgently needed. In this study, we developed an application of DNA vaccine encoding KHV glycoprotein-25 by immersion method to increase survival of common carp against KHV infection. A total of 400 common carp juveniles at 30-day-old were immersed in 1-L water containing 1.3×108CFU/mL of the killed Escherichia coli cells carrying DNA vaccine. Three frequencies and three duration of fish immersion were tested, namely: 1×30 minutes, 1×60 minutes, 1× 90 minutes, 2×90 minutes and 3×90 minutes by interval of 24 hours. Reversetranscription polymerase chain reaction analysis showed that DNA vaccine was successfully expressed in the vaccinated fish. Fish at twenty eight days post vaccination were challenged by injecting 10-4 mL of KHV per fish. The result showed that vaccination by 1×30 minutes immersion allowed 61% of fish survived, and this was significantly higher (p<0.05) compared to control (without vaccination), but it was similar among vaccination treatments (p>0.05). The relative percent survival of vaccinated fish were also similar among treatments (p>0.05). DNA vaccination has increased fish survival about two fold higher compared to unvaccinated fish control (26.67%). Thus, DNA vaccination was effectively delivered by immersion for 1×30 minutes, and this technique can be useful to level up the resistance of common carp juveniles against KHV infection. Keywords: DNA vaccine, KHV, glycoprotein, immersion, common carp
The Use of Synbiotics to Prevent IMNV and Vibrio harveyi Co-Infection in Litopenaeus vannamei OKTAVIANA, ADNI; WIDANARNI, .; YUHANA, MUNTI
HAYATI Journal of Biosciences Vol 21, No 3 (2014): September 2014
Publisher : Bogor Agricultural University, Indonesia

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Abstract

This study evaluated the effects on viral immune responses and bacterial co-infection, of different feeding frequencies of a synbiotic supplemented diet given to Pacific white shrimp (L. vannamei). A synbiotic-supplemented diet was formulated from probiotic Vibrio alginolitycus SKT-bR and prebiotics from sweet potato (Ipomoea batatas L.) oligosaccharide. Pacific white shrimp were fed the synbiotic diet at different frequencies, i.e. daily (P1), twice a week (P2), and once a week (P3) for a 30 day pre-challenge test. After the 30 day feeding period, the shrimps were challenged by intramuscular injection of Infectious Myonecrosis Virus (IMNV) and Vibrio harveyi. The results showed that shrimp treated with a synbiotic-supplemented diet showed  significantly higher growth performance than control groups (P<0.05). Shrimp treated under regime P1 showed the highest values for phenoloxidase (PO) and respiratory burst (RB) parameters compared to shrimp given the other treatments. Following the challenge test, higher survival rate were seen in the P1 treatment group, in comparison to the positive control, and the P1 treatment group showed the highest values in total haemocyte count (THC), PO, and RB.
Effect of Micro-encapsulated Synbiotic at Different Frequencies for Luminous Vibriosis Control in White Shrimp (Litopenaeus vannamei) MUNAENI, WAODE; YUHANA, MUNTI; WIDANARNI, WIDANARNI
Microbiology Indonesia Vol 8, No 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

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Abstract

The aim of this study was to evaluate the effect of micro-encapsulated synbiotic application at different frequencies for luminous disease control in white shrimp (Litopenaeus vannamei). The luminous disease is caused by Vibrio harveyi. In this experiment, a synbiotic which was a combination of the probiotic Bacillus sp. NP5 RfR and the oligosaccharides from sweet potato (Ipomea batatas L.) jago variety was apllied. The synbiotic was encapsulated by spray drying method. The in vivo experiment was conducted by supplementing the shrimp’s diet with the micro-encapsulated synbiotic for 40 days. Treatments included the administration micro-encapsulated synbiotic in different frequencies i.e. once a week (A), twice a week (B), daily (C), and without micro-encapsulated synbiotic (control treatment). The control treatment consisted of positive (K+) and negative (K-) controls. After 30 days period, all of the shrimp were challenged by intramuscular injection of pathogenic V. harveyi RfR at a concentration of 106 CFU ml-1 except the negative control. The treatment C resulted in significantly higher survival rate (SR), specific growth rate (SGR), and immune responses than those of the controls, whereas the feed conversion ratio (FCR) was lower than the controls. In addition, the population of Bacillus sp. NP5 RfR and total bacterial count (TBC) in the intestines increased, whereas the population of V. harveyi RfR and the total vibrio count (TVC) were lower compared to the controls.
Bacillus NP5 Improves Growth Performance and Resistance Against Infectious Myonecrosis Virus in White Shrimp (Litopenaeus vannamei) (Bacillus NP5 Meningkatkan Pertumbuhan dan Ketahanan Terhadap Infeksi Virus Myonecrosis pada Udang Putih (L. vannamei) Widanarni, Widanarni; Yuhana, Munti; Muhammad, Arief
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 19, No 4 (2014): Jurnal Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

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Abstract

Infectious Myonecrosis (IMN) merupakan salah satu penyakit yang sering menyerang udang vaname. Probiotik banyak digunakan pada budidaya udang karena terbukti mampu mengurangi serangan penyakit pada udang. Penelitian ini bertujuan untuk menguji pengaruh pemberian probiotik Bacillus NP5 melalui pakan terhadap kinerja pertumbuhan, respons imun, dan resistensi udang vaname terhadap infeksi Infectious Myonecrosis Virus (IMNV). Udang vaname Litopenaeus vannamei (2.41±0.07 g ekor-1) diberi pakan yang disuplementasi probiotik Bacillus NP5 dengan dosis yang berbeda, 102 CFU.g-1 (A), 104 CFU.g-1 (B), 106 CFU.g-1 (C), dan kontrol tanpa suplementasi probiotik (kontrol negatif, KN; kontrol positif, KP) selama 30 hari dan dengan tiga ulangan untuk masing-masing dosis, kemudian KP, perlakuan A, B, dan C diuji tantang secara intramuskular dengan IMNV (100 µl.ekor-1). Hasil penelitian ini menunjukkan bahwa udang vaname yang diberi pakan dengan suplementasi probiotik mempunyai laju pertumbuhan harian (LPH), rasio konversi pakan (RKP), dan respons imun yang lebih tinggi. Udang tersebut juga mempunyai total hemocyte count (THC) dan resistensi terhadap IMNV yang lebih tinggi dibandingkan kontrol positif. Konsentrasi probiotik 106 CFU.g-1 memberikan hasil terbaik dalam meningkatkan pertumbuhan, respon imun, dan resistensi udang vaname terhadap infeksi IMNV. Kata kunci: probiotik, Bacillus NP5, Litopenaeus vannamei, pertumbuhan, IMNV Infectious Myonecrosis (IMN) is one of the most prevalent white shrimp diseases. Probiotics are widely used in shrimp cultivation because they have been proven to reduce shrimp disease outbreak. This study aimed to observe the effect of oraly administered probiotic Bacillus NP5 on the white shrimps growth performance, immune response, and resistance to Infectious Myonecrosis Virus (IMNV) infection. White shrimp Litopenaeus vannamei (2.41±0.07 g individual-1) were fed with a feed supplemented with different doses of the probiotic Bacillus NP5, i.e. 102 CFU.g-1 (A), 104 CFU.g-1 (B), 106 CFU.g-1 (C), and control without any probiotic (negative control, KN; positive control, KP) for 30 days and with three replications for each dose, then KP, treatment A, B, and C were challenged intramuscularly with IMNV (100 µl.shrimp-1). The results of the study showed that white shrimp fed with the supplemented probiotic had higher Daily Growth Rate (DGR), Feed Conversion Ratio (FCR), and immune response. They also had the higher Total Hemocyte Count (THC) and resistance to IMNV than the positive control. Probiotic with concentration of 106 CFU.g-1 gave the highest value on enhancing growth, immunity, and resistance of white shrimp towards IMNV infection. Key words: probiotic, Bacillus NP5, Litopenaeus vannamei, growth, IMNV
Detection of Luminous Vibrio harveyi in Penaeid Shrimp Through Nested PCR Using Haemolysin Gene Primer SETIAWAN, WAWAN ABDULLAH; WIDYASTUTI, UTUT; YUHANA, MUNTI
HAYATI Journal of Biosciences Vol 22, No 2 (2015): April 2015
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Whiteleg shrimp (Litopenaeus vannamei)  is one of the most important aquaculture commodity in Indonesia. However, the luminous disease primarily caused by Vibrio harveyi bacteria still becomes an obstacle in penaeid shrimp farming, especially in shrimp hatchery. This study was aimed to identify the presence of V. harveyi in L. vannamei through nested PCR using haemolysin gene primer. First, initial primers were designed using V. harveyi VIB 391 haemolysin gene sequence (accession number: DQ640264), flanking the position 133 to 756. This primer pairs were used to identify haemolysin gene in both V. harveyi MR5339 and V. harveyi 275 strain. Sequencing results from each sample showed 99% similarity with haemolysin gene sequence in Genebank. Furthermore, the sequence of V. harveyi MR5339 haemolysin gene was used to design the nested PCR primers. The first primer pairs of nested PCR have successfully amplified the haemolysin gene fragment of all V. harveyi strains samples from position 52 to 405. The second primer pairs of nested PCR have amplified position 204 to 405 where it can detect all of V. harveyi strains used as sample sources in this study. The application of nested PCR technique in this study was able to identify V. harveyi strains at serial dilution of cells density as low as 100 cfu/mL, which is equal to a single cell or at DNA concentration up to 101 fg/µL.
Effects of Microencapsulated Synbiotic Administration at Different Dosages against heavy co-infection of White Spot Disease (WSD) and Vibrio harveyi in Pacific White Shrimp (Litopenaeus vannamei) Yunarty, Yunarty; Yuhana, Munti; Widanarni, Widanarni
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 21, No 4 (2016): Jurnal Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

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White spot disease (WSD) is one of infectious disease in shrimp caused by white spot syndrome virus (WSSV). This study aimed to determine the dosage immunological effects and growth performances of microencapsulated synbiotic (Bacillus NP5 and mannan oligosaccharide) at different dosages on Pacific white shrimp.  The microencapsulated synbiotic   was administered as feed supplementation  against the co-infection of   WSSV and Vibrio harveyi. Synbiotic was encapsulated by spray drying method, further feed supplemented to Pacific white shrimp for 30 days at a  dosages of 0.5% (A), 1% (B), 2% (C) and control treatments, i.e. without any microencapsulated synbiotic administration as positive control (D) and negative control (E). The challenge test was performed on day 30 after feeding supplementation, then the experimental shrimps were injected by WSSV intramuscularly   at the infective dosage of 104 copies.-ml-1. Afterwards,   24 hours after WSSV injection the shrimps were immersed in water contained cells suspension of V. harveyi  at the cells population dosage of 106 CFU-.ml-1. All synbiotic treatments showed better results with the values of Total Haemocyte Count (THC), Phenoloxidase (PO) and Respiratory Burst (RB), were higher (P<0.05) compared to positive control. The specific growth rates (SGR) of A, B and C showed higher than both controls of D and E. The feed conversion ratio (FCR) value of synbiotic treatments were lower (P<0.05) than both controls. However, the administration of microencapsulated synbiotic have not been able to prevent heavy impact of WSSV and V. harveyi co-infection due to lower SR and mortality pattern which continued to increase.  Keywords: Synbiotic, Litopenaeus vannamei, WSSV, Vibrio harveyi, co-infection