GENETIC DIVERSITY OF AMPICILLIN-RESISTANT Vibrio ISOLATED FROM VARIOUS STAGES OF TIGER SHRIMP LARVAE DEVELOPMENT

This research was carried  out to  study  genetic  diversity of ampicillin-resistant Vibrio  from various  stages of tiger  shrimp larvae  (Penaeus Monodon) development from,Tambak Inti Rakyat hatchery, near Labuan, West Java, Indonesia. A total of 25 ampicillin-resistant Vibrio isolates were isolated using thiosulphate  citrate bile-salt  sucrose  agar (TCBS-Agar) and seawater  complete agar (SWC-Agar). Physiological and biochemical characterization showed that the isolates could be grouped into only two species, i.e. V. harveyi from the egg stage; and V. metschnikovii from larvae and post-larval stage (i.e nauplius, zoea, mysis, PLi, PL5, PL,0, and PL,5). These isolates were also present in their respective rearing water  of  each stage  and  some natural feed.  Schizotyping analysis employing restriction endonuclease Noll (5'-GC4GGCCGC) indicated that the isolates could be grouped into  at least  13 different  genotypes.  Therefore,  schizotyping  was  more discriminative than physiological characterization. This study showed that particular groups of Vibrio colonized all stages of shrimp larvae and demonstrated closed phylogenetic relationship. These groups of Vibrio might be  the dominant microbiota which could suppress the development of other Vibrio including the pathogenic Vibrio. Key words : Shrimp/ampicillin-resistant K/fcno/schizotypin

PERKEMBANGAN Aeromonas Hydrophila PADA BERBAGAI MEDIA KULTUR

Bakteri A. hydrophila merupakan bakteri patogen fakultatif anaerob dan umum ada di setiap perairan. Bersifat oportunistik atau tidak menjadi berbahaya jika dalam kondisi budidaya yang baik, akan tetapi bila kondisi budidaya buruk maka akan dapat menyebabkan kematian massal baik ukuran benih maupun induk dalam waktu yang relatif singkat sehingga mengakibatkan kerugian yang cukup besar. Namun demikian keberadaan bakteri patogen ini pada berbagai media hidup belum teramati dengan baik. Tujuan penelitian ini adalah untuk mengetahui kepadatan A. hydrophila pada berbagai media hidup. Penelitian ini bersifat eksperimental laboratoris yang dilakukan dalam tiga tahap penelitian. Tahap pertama adalah pengujian perkembangan A. hydrophila secara in vitro pada berbagai media kultur agar/broth. Tahap kedua adalah pengujian perkembangan A. hydophila pada media budidaya tanpa ada ikan dan tahap ketiga adalah perkembangan A.hydrophila di media budidaya ikan mas (Cyprinus carpio). Hasil yang didapatkan dalam penelitian ini adalah bahwa fase eksponensial A. hydrophila pada media kultur TSB terjadi pada jam ke-16 hingga jam ke-22 dengan peningkatan jumlah mencapai puncaknya pada jam ke-22 dengan kepadatan bakteri mencapai 1.66x10 cfu/mL. Faktor yangmenyebabkan peningkatan kepadatan A. hydrophila disebabkan adanya nutrien yang ada di media budidaya. Puncak perkembangan/ peningkatan kepadatan A.hydrophila pada media budidaya ikan Mas (Cyprinus carpio) adalah hari ke-8 yakni mencapai 5x10 cfu/mL dan mengakibatkan SR ikan Mas selama masa penelitan hanya 36,67%. Kata kunci: Perkembangan Aeromonas hydrophila, media agar, media budiday

Growth performance, immune response, and resistance of Nile tilapia fed paraprobiotic Bacillus sp. NP5 against Streptococcus agalactiae infection

ABSTRACT   This study was aimed to evaluate the effectiveness of Bacillus sp. NP5 paraprobiotic administration through commercial feed on growth performance, immune response, and resistance of Nile tilapia against Streptococcus agalactiae infection. Bacillus sp. NP5 paraprobiotic was produced through heat-inactivation at 95°C for 1 h, then performed a viability test on tryptic soy agar (TSA) media and incubated for 24 hours. Paraprobiotics could be used whether the bacteria did not grow on the TSA media. This study used a completely randomized design, containing three treatments with five replications, i.e. 1% (v/w) probiotic addition, 1% (v/w) paraprobiotic addition, and no addition of probiotic or paraprobiotic (control). The experimental fish were reared for 30 days. On day 31 of rearing, fish were challenged with S. agalactiae (107 CFU/mL) through intraperitoneal injection route, while the negative control was injected with PBS. This study results significantly improved growth performances and immune responses (P<0.05), compared to control after 30 days of probiotic and paraprobiotic Bacillus sp. NP5 administration. After challenge test, increased immune responses in probiotic and paraprobiotic of Bacillus sp. NP5 treatment had higher survival rates (P<0.05) than positive control. The administration of Bacillus sp. NP5 probiotic and paraprobiotic through commercial feed were effective in increasing growth performance, immune response, and resistance of Nile tilapia against S. agalactiae infection. Keywords: Bacillus sp. NP5, Nile tilapia, paraprobiotic, Streptococcus agalactiae   ABSTRAK   Penelitian ini bertujuan mengevaluasi efektivitas pemberian paraprobiotik Bacillus sp. NP5 melalui pakan dalam meningkatkan kinerja pertumbuhan, respons imun, dan resistansi ikan nila terhadap infeksi Streptococcus agalactiae. Proses pembuatan bakteri paraprobiotik yaitu Bacillus sp. NP5 diinaktivasi panas pada suhu 95°C selama 1 jam, dilanjutkan dengan pengujian viabilitas dengan menyebarkannya pada media tryptic soy agar kemudian diinkubasi selama 24 jam. Jika bakteri tidak tumbuh, maka paraprobiotik dapat digunakan. Penelitian ini menggunakan rancangan acak lengkap dengan tiga perlakuan dan lima ulangan, yaitu penambahan probiotik 1% (v/w), penambahan paraprobiotik 1% (v/w), dan tanpa penambahan probiotik atau paraprobiotik (kontrol). Ikan perlakuan dipelihara selama 30 hari dan pada hari ke-31, ikan diuji tantang dengan S. agalactiae (107 CFU/mL) melalui injeksi intraperitoneal, sementara perlakuan kontrol negatif diinjeksi dengan PBS. Hasil penelitian setelah 30 hari pemberian probiotik dan paraprobiotik Bacillus sp. NP5 menunjukkan kinerja pertumbuhan dan respons imun yang meningkat signifikan (P<0.05) dibandingkan dengan kontrol. Pascauji tantang, peningkatan respons imun pada perlakuan probiotik dan paraprobiotik Bacillus sp. NP5 menunjukkan tingkat kelangsungan hidup yang lebih tinggi (P<0.05) dibandingkan kontrol positif. Pemberian probiotik dan paraprobiotik Bacillus sp. NP5 melalui pakan dapat meningkatkan kinerja pertumbuhan, respons imun, dan resistansi ikan nila terhadap infeksi Streptococcus agalactiae. Kata kunci: Bacillus sp. NP5, ikan nila, paraprobiotik, Streptococcus agalactia

The presence of disease in growing spiny lobster Panulirus spp. in floating net cages at Tanjung Putus Island, Lampung

Spiny lobster is a member of crustaceans that has high economic value. The demand for export markets in Asian, European and American countries as well as locally, is quite high with an expensive selling price. However, there are obstacles that often occur in spiny lobster rearing cultivation, namely infectious diseases that will cause high mortality. The purpose of this study was to investigate infectious diseases in rearing spiny lobster Panulirus spp. that were kept in floating net cages at Tanjung Putus, Lampung. The research method involved sampling five spiny lobsters in floating net cages, observation of clinical symptoms, bacterial isolation, identification of bacteria, detection of Milky Hemolymph Disease of Spiny Lobster (MHD-SL) through polymerase chain reaction (PCR) analysis and histopathology. The sampling location was at PT. Saibatin Perikanan Indonesia. The bacteria Vibrio parahaemolyticus and Vibrio alginolyticus were detected. Five lobsters were infected by MHD-SL, characterized by milky white hemolymph. All five lobster samples were positively infected with Milky Hemolymph Disease of Spiny Lobster (MHD-SL) through PCR analysis. Histopathological observations showed pathological microanatomical changes in the lobster hepatopancreas tissue, indicating that the tissue changes, in the form of encapsulation and infiltration were due to Rickettsia-like bacteria (RLB) infection. The management of spiny lobster stocking density, maintenance biosecurity, and increased lobster immunity must be implemented in order to prevent MHD-SL disease in floating net cages.   Keywords: bacteria, histopathology, MHD-SL, Panulirus spp.   ABSTRAK   Lobster merupakan anggota crustacea yang memiliki nilai ekonomis yang tinggi. Permintaan pasar ekspor pada negara Asia, Eropa, dan Amerika maupun lokal cukup tinggi dengan nilai jual yang mahal. Akan tetapi, terdapat kendala yang sering terjadi pada budidaya pembesaran lobster yaitu infeksi penyakit yang akan menyebabkan tingginya mortalitas Tujuan penelitian ini adalah untuk menginvestigasi penyakit infeksi pada pembesaran lobster Panulirus spp. yang dipelihara di keramba jaring apung Tanjung Putus, Lampung. Metode penelitian ini meliputi pengambilan sampel lobster sebanyak lima ekor dalam karamba jarring apung (KJA), pengamatan gejala klinis, isolasi bakteri, identifikasi bakteri, deteksi Milky Hemolymph Disease of Spiny Lobster (MHD-SL) melalui analisis Polymerase Chain Reaction (PCR), dan histopatologi. Lokasi pengambilan sampel di PT. Saibatin Perikanan Indonesia, Lampung. Terdapat bakteri Vibrio parahaemolyticus dan V. alginolyticus. Lima ekor lobster terinfeksi oleh MHD-SL yang ditandai dengan hemolim berwarna putih susu. Lima sampel lobster positif terinfeksi penyakit Milky Hemolymph Disease of Spiny Lobster (MHD-SL) melalui analisis PCR. Hasil pengamatan histopatologi terdapat perubahan mikroanatomi patologi pada jaringan hepatopankreas lobster menunjukkan bahwa adanya perubahan jaringan berupa enkapsulasi dan infiltrasi karena adanya infeksi Rickettsia-like bacteria (RLB). Pengelolaan kepadatan penebaran lobster, biosekuritas pemeliharaan, dan peningkatan imunitas lobster harus ditingkatkan untuk mencegah penyakit MHD-SL di KJA.   Kata kunci: bakteri, histopatologi, MHD-SL, Panulirus spp

Total Hemocytes, Glucose Hemolymph, and Production Performance of Spiny Lobster Panulirus Homarus Cultured in the Individual Compartments System

Cannibalism can cause high mortality in a lobster culture. To prevent cannibalism, an artificial shel-ter is needed like the individual compartments system (ICS) to ensure that there is no contact bet-ween lobsters. This study aimed to evaluate the application of ICS on total hemocytes glucose hemo-lymph, production performance, and determine the shape of more effective ICS to minimize stress level of lobster. The treatments used for various application shape of ICS was tubular ICS, triangle ICS, square ICS, and shelter PVC pipes as control. Lobster maintenance was conducted in outdoor for 60 days. The results indicated that physiological responses with square ICS treatment were more effective to reduce stress level compared with other treatments (p<0.05). Application of ICS was more optimal to improved SR (p<0.05) and growth of lobster (p>0.05). Overall, treatment of square ICS was opti-mal related to production performance i.e. SR 88.89±5.24%, SGR 0.61±0.49%/day, with harvest size of total length 137.31±8.11 mm/ind, weight 58.83±4.78 g/ind, and FCR 22.71±1.72. Application of ICS in lobster culture was effective to reduce stress level as indicated from total hemocytes and gluco-se hemolymph, and support optimal production performance. Lobster culture using a square ICS sha-pe was effective to minimize stress level, compared with tubular ICS and triangle ICS

CONTROL OF Vibrio alginolyticus INFECTION IN ASIAN SEA BASS Lates calcarifer USING AMBON BANANA PLANT POWDER Musa paradisiacal THROUGH THE FEED

Asian sea bass is one of economically important commodities in aquaculture. However, its culture often challenged by vibriosis infection which resulted in lower production. The study aimed to evaluate the effectiveness of the ambon banana plant powder application in feed as an effort to control the disease caused by Vibrio alginolyticus infection. Asian sea bass with total length of 7.48 ± 0.45 cm. Five different treatments were applied in this experiment, i.e. positive control, negative control, prevention, curative, and antibiotic application. The challenge test is carried out on the 15th day. Challenge test was performed with intramuscularly injection of V. alginolyticus at the cell density of 106 CFU/fish. The results showed the amount of feed consumption after the challenge test was highest in the curative and antibiotic control. However, these results were not significantly different (P>0.05) among preventive and curative treatments but were significantly different from controls (P<0.05). Fish treated with supplemented feed showed the highest total erythrocytes, total leukocytes, hemoglobin, respiratory burst, and lysozyme activity compared to controls. IL-1b gene expression increased after the challenge test with the highest level of expression in the curative treatment. It can be concluded that the administration of ambon banana plant powder (3 g/100 g diet) could control of V. alginolyticus infection in Asian sea bass and resulted 70.00% survival rate on prevention and 83.33% on curative treatments

Isolation and Characterization of Lactic Acid Bacteria from Inasua

Inasua is a traditionally product of wet salt fish fermentation produced by Teon, Nila and Serua (TNS) Communities in Central Maluku, Indonesia. The community made this fermented fish to anticipate the lean time when fisherman could not go to sea.  The  fish that used as inasua raw material is demersal fishes that live around coral reefs, such as Samandar fish (Siganatus guttatus), Gala-gala fish (Lutjanus sp.) and Sikuda fish (Lethrinus ornatus). The objective of the research was to isolate and characterize of bacterial indigenous in  Inasua from three producers in Seram Island. The measurement of pH from inasua samples were 5.9, 5.0 and 5.8, respectively. The highest number of lactic acid bacteria was found from  Gala – gala inasua was 2,5x107 cfu/g sample. Isolation of all isolates bacteria from inasua showed that a total of 7 isolates of bacteria was obtained  from Samadar inasua, 9 isolates from  Gala-gala inasua, and 7 isolates from  Sikuda inasua.  From a total of 23 isolates, only 6 isolates had characteristic as lactic acid bacteria that were Gram  positive, negative catalase, and cocci shape. The microscopic characteristics  of the isolates are coccid in pairs or uniforms which combine to form tetrads. Carbohydrate utilization test  of selected isolate by using API 50 CHB kit indicated that 13 carbohydrates are fermented by these isolates  after incubation for 48 hours. The research  was concluded that the dominant bacteria in inasua sample  is  cocci-lactic acid bacteria.Keywords : fermented fish, inasua, lactic acid bacteria, MRSA mediu

POTENCY OF VIBRIO ISOLATES FOR BIOCONTROL OF VIBRIOSIS IN TIGER SHRIMP (PENAEUS MONODON) LARVAE

This study was carried out to obtain Vibrio isolates able to function as biocontrol of vibriosis in shrimp hatchery. Thirty one Vibrio isolates were isolated from tiger shrimp larvae and hatchery environments, i.e. Labuan, Pangandaran, and Lampung, Indonesia. Pathogenic  V. harveyi MR5339 was obtained from Maros, South-Sulawesi and was made as a rifampicin resistant mutant (RFR) to screen for those 31 Vibrio isolates in in vitro assays and to allow us to monitor their presence in shrimp larvae and larval rearing water. Almost all Vibrio  isolates could inhibit the growth of pathogenic V. harveyi MR5339 RFR. SKT-b isolate from Skeletonema was the most effective to inhibit the growth of V. harveyi MR5339 Rf* and significantly reduced larval mortality in pathogen challenge assays. These prospective biocontrol bacteria, at concentration of 10" CFU/ml, did not show pathogenicity to shrimp larvae. SKT-b was Gram negative, short rod-shape, exhibited yellow colonies on TCBS and swarming on SWC-agar media, motile, utilized glucose and sucrose but not lactose: produced extra-cellular protease and amylase, but did not produce chitmase. Partial sequencing of 16S-rRNA gene SKT-b showed SKT-b similarity to Vibrio alginofyticus. Keywords: shrimp larvae / biocontrol bacteria / vibriosis