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Phenotypic and Genotypic Comparison of Vibrio in Seawater Fish from Batam and Mataram, Indonesia

Journal of Agricultural Science and Technology(B) 2015年第8期5卷 561-566页

作者： Rio Aditya Kumiawan kumiasih Ministry of Marine Affairs and Fisheries Republic of Indonesia Fish Quarantine and Inspection Agency Jayapura Papua 99352Indonesia Faculty of Veterinary Medicine Gadjah Mada University Yogyakarta 55281 Indonesia

Grouper and snapper are the potential fishery commodity in Indonesia with a high economic value, as well as an export commodity. A common disease in grouper and snapper aquaculture is vibriosis. Vibriosis is a disease caused by bacteria of the genus Vibrio. The aim of study was to compare between phenotypic and genotypic identification of Vibrio isolated from Batam and Mataram, Indonesia. Bacteria were isolated from anterior kidney and eye of fish, then grown in thiosulfate-citrate-bile salts-sucrose （TCBS） and incubated in room temperature （25-28 ~C） for 24 h, and identified using morphology and biochemical test. Bacterial isolates were extracted, amplified and sequenced on 16S rRNA region. Phylogenetic tree of bacteria was constructed using neighbor-joining and maximum-parsimony methods. The phenotypic identification was found six isolates of Vibrio from Batam, such as K alginolyticus, V. carchariae, K damselae, V. fluvialis, V. furnissii and K parahaemolyticus. Three isolates were found from Mataram, such as 1I. alginolyticus, V. carchariae and V. fluvialis. Blast analysis showed isolates of V. alginolyticus_btm and V. carchariae_btm homolog to V. parahaemolyticus strain DAHMV3; isolates of V. damselae_btm and K alginolyticus_mtr homolog to V. neocaledonicus strain MS1; isolates of V. parahaemolyticus__btm and V. furnisii_btm homolog with Photobacterium damselae subsp, damselae strain： 04Ya311 and isolate of K fluvialis_mtr homolog to V. azureus strain MMRF532, respectively. All phenotypic identification was not supported by molecular identification on 16S rRNA region. It was suggested that phenotypic identification should be supported by molecular examination, especially in identification of Vibrio species.

关键词： Vibrio phenotype genotype 16S rRNA.

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Concentration of Enrofloxacin Residue from Tilapia （Oreochromis niloticus） Muscular That Infected by Aeromonas salmonicida

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Journal of Agricultural Science and Technology(A) 2014年第9期4卷 750-754页

作者： kumiasih Fera Aryanti Department of Pathology Faculty of Veterinary Medicine Gadjah Mada University 560862 Indonesia

Aim of this research was to find out the concentration of enrofloxacin residue in tilapia meat for several weeks after antibiotic treatment. Twenty seven tilapia fishes were divided into three groups. The first group was not infected and treated, the second group was infected with A. salmonicida subsp, smithia and the third group was infected with A. salmonicida subsp. achromogenes intramuscularly. Six days after infection, treatment was carried out using Baytril administered orally for the second group and intramuscularly for the third group during five days. At the 1 st, 4th and 8th week after the treatment, Three fish were taken from each group to be analyzed for its concentration of enrofloxacin residue by diffusion on Mueller Hinton Agar （MHA） method and quantitatively using high performance liquid chromatography （HPLC） method. The MHA test showed the formation of inhibition zone, at the 1 st week and 4th week after the treatment, while at 8th week after treatment did not show inhibition zone. The HPLC test on enrofloxacin residual concentration in tilapia infected with A. salmonicida subsp, smithia （second group） at the 1st, 4th and 8th week after treatment showed the average of 33.0, 6.10 and 0.0021 μg/g of enrofloxacin residue level. While in tilapia infected with A. salmonicida subsp, achromogenes and treated with enrofloxacin intramuscularly （third group） showed the average of residue level 35.79, 2.18 and 0.00065 μg/g. In conclusion, the residue of enrofloxacin was still high concentration until the fourth week after treatment in the second and third groups. Based on Indonesian National Standards and Rules, the maximum limit of enrofloxacin residue is 0.01 μg/g. The concentration of enrofloxacine residue was very low and the concentration of enrofloxacin residue collected from tilapia using orally and intramuscularly method of treatment was not different.

关键词： Residue enrofloxacin Aeromonas salmonicida subsp smithia subsp achromogenes.

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Phylogeny and Histopathology of Streptococcus iniae from Indonesia

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Journal of Agricultural Science and Technology(B) 2015年第2期5卷 135-140页

作者： Titis Candra Dewi kumiasih Surya Amanu Rini Widayati Fish Quarantine and Inspection Agency Semarang 50129 Indonesia Department of Pathology Faculty of Veterinary Medicine Gadjah Mada University Yogyakarta 560862 Indonesia Department of Microbiology Faculty of Veterinary Medicine Gadjah Mada University Yogyakarta 560862 Indonesia Department of Biochemistry Facul(y of Veterinary Medicine Gadjah Mada University Yogyakarta 560862 Indonesia

Streptococcus iniae has been detected in several regions in Indonesia, such as Bali, Jambi and Papua. Outbreak of meningoencephalitis and septicemia was still often found in Papua. The aim of study was to compare phenotype, genotype and histopathology of S. iniae from Papua. Bacteria was identified using morphological and biochemical tests. It was also extracted, amplified and sequenced to see genotypic characters. The primer used were 5＇-AGAGTTTGATCCTGGCT-3＇（24F） and 5＇-AAGGGAGGTGATCCAGCCGCA-3＇（1540R） in 16S rRNA region. Result of sequencing was analyzed by the neighbor joining and maximum parsimony methods. The identification result of isolate from Papua was S. iniae. The amplification result was a sharp band--l,500 bp band in 16S rRNA region. The phylogenetic tree showed that isolate from Papua was closely related to S. iniae strains CMS005 from Guangxi China. Bacterial clumps of S. iniae was firstly found in the blood vessel of liver at the 3rd day after infection and then caused the inflammation to spleen, heart, brain, kidney and gut at the 7th day post infection.

关键词： Phenotype genotype phylogeny.

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