Isolation and Identification of Aeromonas spp. from Diseased African Catfish (Clarias sp.) in Ngawi Regency

African catfish (Clarias sp.) is one of important freshwater fish which prefer consumed by people due to many advantages. Aeromonas sp. bacteria are dangerous patogen for freshwater fishes. This objective of the research was to isolate, identify and determine pathogenicity of Aeromonas sp. bacteria from African catfish from District Ngawi. The samples of catfish with 20 - 25 cm length showing clinical signs disease were obtained from three sub districts. Bacteria were isolated from kidney and inoculated into GSP medium. Characterization and identification through morphology of bacterial colonies, cells and biochemical test. Postulate Koch was conducted to verify abaility to couse disease. Pathogenicity was analyzed by determination of value of Lethal Dosage-50 on catfish on 7 - 9 cm length. The result showed that the disease symptoms of fish infected by the bacteria were skin ulcer, abdominal swelling and kidney damage. Fifteen bacterial isolates were collected which five, six and four isolates were from Kecamatan Karang Jati, Geneng dan Paron.sub-district respectively. The result showed 12 isolates (GKJ1, GKJ3, GKJ4, GGN1, GGN2, GGN3, GGN4, GGN5, GGN6, GPR2, GPR3 and GPR4) were identified as Aeromonas hydrophila. Three isolates (GKJ2, GKJ5 and GPR1) were identified as A. salmonicida. Isolate of A. hydrophila GKJ1, GKJ4, GGN2, GGN5, GPR2 and GPR4 were virulent to African catfish with LD50 values of 1,55 x 105, 3,89 x 105, 7,24 x 105, 2,39 x 105, 6,61 x 104 and 1,95 x 104 cfu/fish

Cloning ORF2 Membrane Protein of Koi Herpesvirus Lake Toba, Indonesian Isolate

Koi herpesvirus (KHV) caused significant morbidity and mortality in koi and common carp. KHV which showed strong antigenic property implied that KHV virion or proteins may be used as antigen to raise antibody or vaccine to increase the resistance. The objectives of this research were to (i) clone KHV membrane protein ORF2, (ii) analysis on immunogenicity, and (iii) genetic tracing. Based on genbank data, one pair of primers was designed to amplify KHV ORF2. The KHV ORF2 can be amplified using infected fish DNA which originally from Toba Lake, Sumatera, Indonesia. The KHV ORF2 composed of 699 nucleotides encoded for 292 amino acids. BLAST analysis showed that KHV ORF2 had 100% homology with KHV-J and KHV0301 strains from Japan; 98 and 91% homology on nucleotides and amino acids respectively with both KHV-U strain from Unites State and KHV-I strain from Israel. KHV in Indonesia was most likely to have originated from Japan via spreading directly or not directly to China or Hongkong. Based on T- and B-cell epitopes prediction, membrane protein ORF2 was proposed has a potency to be used in development vaccine and immunodetection. Key words: genetic tracing, koi herpesvirus (KHV), membrane protein, ORF

Cloning ORF2 Membrane Protein of Koi Herpesvirus Lake Toba, Indonesian Isolate

Koi herpesvirus (KHV) caused significant morbidity and mortality in koi and common carp. KHV which showed strong antigenic property implied that KHV virion or proteins may be used as antigen to raise antibody or vaccine to increase the resistance. The objectives of this research were to (i) clone KHV membrane protein ORF2, (ii) analysis on immunogenicity, and (iii) genetic tracing. Based on genbank data, one pair of primers was designed to amplify KHV ORF2. The KHV ORF2 can be amplified using infected fish DNA which originally from Toba Lake, Sumatera, Indonesia. The KHV ORF2 composed of 699 nucleotides encoded for 292 amino acids. BLAST analysis showed that KHV ORF2 had 100% homology with KHV-J and KHV0301 strains from Japan; 98 and 91% homology on nucleotides and amino acids respectively with both KHV-U strain from Unites State and KHV-I strain from Israel. KHV in Indonesia was most likely to have originated from Japan via spreading directly or not directly to China or Hongkong. Based on T- and B-cell epitopes prediction, membrane protein ORF2 was proposed has a potency to be used in development vaccine and immunodetection

Distribution of Ammonium-Oxidizing Bacteria in Sediment with Relation to Water Quality at the Musi River, Indonesia

The Musi River is located in the southern Sumatra, Indonesia. Most of activities, i.e. agricultural, industrial, and urban activities are considered as being major sources of chemicals and nutrients with their waste products effluent into the river. Nitrification, the microbial oxidation of ammonia to nitrite and nitrate, occurs in a wide variety of environments and naturally remove anthropogenic N pollution. The purpose of this research was to determine of distribution of ammonium-oxidizing bacteria (AOB) in sediment with relation to water quality at the Musi river area. This study was conducted on rainy and dry season 2016 at five sampling sites from the freshwater to seawater at high and low tide conditions, the sampling sites are station St1 (Gandus), station St2 (Palembang city), station St3 (Upang), station St4 (Sungsang), and station St5 (Sea). Sediment samples were collected from the surface layer by using an Ekman grab. Some water quality such as salinity, temperature, pH, and dissolved oxygen (DO) were directly analyzed in the field, while other water quality such as NH4-N, NO2-N, and NO3-N were analyzed in the laboratory. The Density of AOB was determined by the most probable number of (MPN) method. The PCA was used to correlate variations of the AOB with physicochemical properties using software Xlstat. The results showed that the physicochemical properties had a range of salinity of 0 to 20 ppt, temperature of 29.21 to 31.82oC, pH of 4.88 to 7.93, DO of 3.44 to 11.33 mg/l, NH4-N in sediment of 0.04 to 0.87 mg/l, NO2-N in sediment of 0.01 to 1.77 mg/l, NO3-N in sediment of 0.09 to 2.08 mg/l. The density of AOB ranged from 7.2 x 102 to 6.1 x 103 cells/g sediment. Principal component analyses showed that temperature, pH, DO, and concentrations of nutrient contributed to density of AOB

Study of the Effects of Carboxymethyl Chitosan on the Non-specific Defense System in the Carp (Cyprinus Carpio)

Carp (Cyprinus carpio) is a freshwater fish with a high economic value, but very susceptible to diseases. One of effort to increase the productivity is by enhancing non-specific defense system. The purpose of this study is to determine the effect of carboxymethyl chitosan on enhancement non-specific defense system of carp. Carboxymethyl chitosan was obtained by alkylation process in which monochloroacetic acid in alkaline conditions was added. Carboxymethyl chitosan was given to carps at dosages of 30 μg/g, 75 μg/g and 105 μg/g, by intra muscular injection respectively. Seven and 14 days after administration of carboxymethyl chitosan, measurements of non-specific immune system parameters were done. The results showed that, administration of carboxymethyl chitosan on carps affected the phagocytic activity and lymphocytes counts. However, carboxymethyl chitosan did not give any effect to NBT activity, hematocrit, number of erythtocytes and leukocytes, monocytes and neutrophil counts in blood as well

PENGEMBANGAN METODE LOOP-MEDIATED ISOTHERMAL AMPLIFICATION OF DNA DAN APLIKASINYA UNTUK DETEKSI KOI HERPES VIRUS PADA BEBERAPA JENIS IKAN

The purposes of this experiment were to establish loop-mediated isothermal amplification of DNA ( LAMP) method and its application to observe the presence and duration of koi herpes virus ( KHV) in freshwater fishes after infection. The infection was carried out using 4-6 cm length of java barb (Barbodes gonionotus), grass carp (Ctenoparingodon idella), gold fish (komet) (Carassius auratus), tambaqui (Colossoma macropomum) and koi (Cyprinus carpio koi) fishes. Fishes were infected intraperitoneally with KHV inoculums and 2 fishes were sampled everyday. DNA was extracted from gill and used for diagnosis with LAMP assays. The result of this experiment showed LAMP assay can be established and gave 100 times more sensitive than conventional PCR assay. Koi was confirmed as a host of KHV. Java barb, tambaqui, gold fish and grass carp can serve as vector for KHV and the presence of KHV was detected in java barb and tambaqui for 4 days, goldfish and grass carp for 5 days after infection

Physiological, biochemical and HSP70 and HSP90 gene expression profiles of tropical abalone Haliotis squamata in response to Vibrio alginolyticus infection

Vibrio spp. have been known responsible for fish diseases in marine and brackish‐water systems in the tropics regions. Heat shock proteins are a highly conserved protein group that is known for its rapid response to environmental stresses, including infection. This study aimed to investigate physiological and biochemical responses of tropical abalone Haliotis squamata to Vibrio alginolyticus infection. Abalones were infected with V. alginolyticus by intramuscular injection at a dose of 105,106,107 cfu/abalone. The expression of HSP70 and HSP90 genes, the activity of superoxide dismutase, phenol oxidase and catalase enzymes, histology, falling and mortality were observed at 12, 24, 48, 72, and 96 hours post‐infection (hpi). The different expression of HSPs was found in this study. While the expression of HSP70 was downregulated after infection, the expression of HSP90 was upregulated at 12 hpi and followed by downregulated after 24 hpi for 106 cfu infection, but expressed at a normal level for 105 infection treatment. The expression ofsuperoxide dismutase and catalase increased within 12 hpi, and the expression of phenol oxidase increased after 24 hpi. V. alginolyticus is virulent with LD50 of less than 105 cfu on H. squamata with an average weight of 5.13 g, and caused enlargement of hemolymph sinus and development intraepithelial and intramuscular abscesses

Analysis of Htra Gene from Zebrafish (Danio Rerio)

HtrA which is characterized by the combination of a trypsin-like catalytic domain with at least one C-terminal PDZ domain is a highly conserved family of serine proteases found in a wide range of organisms. However the identified HtrA family numbers varies among spesies, for example the number of mammalian, Eschericia coli, fruit fly-HtrA family are 4, 3 and 1 gene respectively. One gene is predicted exist in zebrafish. Since no complete information available on zebrafish HtrA, in this paper zebrafish HtrA (zHtrA) gene was analyzed. The zHtrA is belonged to HtrA1 member and predicted encodes 478 amino acids with a signal peptide, a IGF binding domain, a Kazal-type inhibitor domain in the up stream of HtrA-bacterial homolog. At the amino acid sequence the zHtrA1 showed the 69%, 69%, 68%, 54% and 54% with the rat HtrA1, mouse HtrA1, human HtrA1, human HtrA3 and mouse HtrA4 respectively. The zHtrA1 is firstly expressed at 60 hpf and mainly in the vertebral rudiments in the tail region