. TRIYANTO
Department of Fisheries, Faculty of Agriculture, Gadjah Mada University

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Selective Media for In Vitro Activity Evaluation of Bacterial Biocontrol Against Pathogenic Vibrio ISNANSETYO, ALIM; MUHTADI, .; ISTIQOMAH, INDAH; NITIMULYO, KAMISO HANDOYO; TRIYANTO, .
HAYATI Journal of Biosciences Vol 18, No 3 (2011): September 2011
Publisher : Bogor Agricultural University, Indonesia

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Abstract

In vitro activity test is a critical evaluation to screen the potential biocontrol agent. We developed a selective medium for quantitative in vitro activity evaluation of bacterial biocontrol agents against pathogenic Vibrio in aquaculture. Sensitivity test of bacterial biocontrol and Vibrio spp. to nine antibiotics showed that oxytetracycline inhibited the growth of Vibrio spp., but did not inhibit the growth of the bacterial biocontrol. This selective inhibition activity of oxytetracycline suggested this antibiotic might be supplemented to establish a selective medium. The MIC of oxytetracycline against V. harveyi, V. parahaemolyticus, and V. algosus were 120, 250, and 120 mg/ml, respectively. These concentration did not inhibit the growth of bacterial biocontrols. Therefore, oxytetracycline was supplemented at 250 µg/ml in Zobell agar medium. This medium was used as a selective medium to enumerate the density of bacterial biocontrol. In vitro activity test of bacterial biocontrol (RLP1) against V. parahaemolyticus showed that strain RLP1 at density of 104 and 106 cells/ml was able to kill V. parahaemolyticus during 6 h incubation. At lower density, 102 cells/ml, this bacterial biocontrol agent was able to kill the pathogenic Vibrio during 12 h incubation. This study discovered a selective medium for the bacterial biocontrol and Vibrio spp. and provided the results of its application in the evaluation of in vitro activity of a bacterial biocontrol agent against V. parahaemolyticus. The results also revealed that strain RLP1 is a potential bacterial biocontrol against vibriosis in marine aquaculture.
In vitro activity test is a critical evaluation to screen the potential biocontrol agent. We developed a selective medium for quantitative in vitro activity evaluation of bacterial biocontrol agents against pathogenic Vibrio in aquaculture. Sensitivity test of bacterial biocontrol and Vibrio spp. to nine antibiotics showed that oxytetracycline inhibited the growth of Vibrio spp., but did not inhibit the growth of the bacterial biocontrol. This selective inhibition activity of oxytetracycline sug ISNANSETYO, ALIM; MUHTADI, .; ISTIQOMAH, INDAH; NITIMULYO, KAMISO HANDOYO; TRIYANTO, .
HAYATI Journal of Biosciences Vol 18, No 3 (2011): September 2011
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Original Source | Check in Google Scholar

Abstract

In vitro activity test is a critical evaluation to screen the potential biocontrol agent. We developed a selective medium for quantitative in vitro activity evaluation of bacterial biocontrol agents against pathogenic Vibrio in aquaculture. Sensitivity test of bacterial biocontrol and Vibrio spp. to nine antibiotics showed that oxytetracycline inhibited the growth of Vibrio spp., but did not inhibit the growth of the bacterial biocontrol. This selective inhibition activity of oxytetracycline suggested this antibiotic might be supplemented to establish a selective medium. The MIC of oxytetracycline against V. harveyi, V. parahaemolyticus, and V. algosus were 120, 250, and 120 mg/ml, respectively. These concentration did not inhibit the growth of bacterial biocontrols. Therefore, oxytetracycline was supplemented at 250 ?g/ml in Zobell agar medium. This medium was used as a selective medium to enumerate the density of bacterial biocontrol. In vitro activity test of bacterial biocontrol (RLP1) against V. parahaemolyticus showed that strain RLP1 at density of 104 and 106?cells/ml was able to kill V. parahaemolyticus during 6 h incubation. At lower density, 102 cells/ml, this bacterial biocontrol agent was able to kill the pathogenic Vibrio during 12 h incubation. This study discovered a selective medium for the bacterial biocontrol and Vibrio spp. and provided the results of its application in the evaluation of in vitro activity of a bacterial biocontrol agent against V. parahaemolyticus. The results also revealed that strain RLP1 is a potential bacterial biocontrol against vibriosis in marine aquaculture.
Gene Cloning and Protein Expression of Koi Herpesvirus ORF25 Murwantoko, .; Fusiyanto, Cahya Kurnia; Triyanto, .
HAYATI Journal of Biosciences Vol 23, No 3 (2016): July 2016
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Koi herpesvirus (KHV) caused significant morbidity and mortality in koi and common carp (Cyprinus carpio). Glycoprotein has been used for vaccine development as sub unit vaccine against many viruses.?KHV ORF25?is one of koi herpesvirus genes which encode a glycoprotein. The objectives of this research are to clone gene?KHV ORF25and express its protein. The common carp showing necrosis and white patches of gill which was collected from Magelang was used in this research. Primers were designed to amplify partial ORF25 based on KHV J strain.?KHV ORF25?was successfully amplified and cloned in pET32a. Sequence analysis showed that this?KHV ORF25?has 99% homology with the sequences of KHV genotype KHV-J, KHV-I, and KHV-U. This ORF was predicted has 3, 23, and 8 B-cell epitopes based on Emini scale, Karplus and Schulz scale, and ElliPro respectively. The KHV ORF25 recombinant protein has been successfully produced in?Escherichia coli?as an insoluble protein with approximately 45?kDa in size. The high protein production was achieved when the protein induction was done at bacterial density at OD600?as 1.0 with 1-mM isopropyl ?-D-1-thiogalactopyranoside (IPTG) and incubated at 37?C for 18?hours. The protein predicted has immunogenicity and the potency as a vaccine is needed to be evaluated.