Peromonas Hydrophila Lab Report

2. MATERIALS AND METHODS

2.1 The isolation of Aeromonas hydrophila from various environments using phenospecies method.

There were three groups of bacteria used in the present research. The first group was isolated from the gut of diseased fish which obtained from Balai Riset Perikanan Budidaya Air Tawar (Research Institute of Freshwater Aquaculture) Sempur Bogor and Sukabumi. The second group was isolated from healthy fish intestine. The third group was isolated from the fish ponds from West Java Indonesia cities such as Tasikmalaya, Garut, Bandung, Indramayu and Sukabumi. Each colony of A. hydrophila grew white and the middle part was bright yellow in the Rimler-Shoots medium + novobiocin (SNI 7303, 2009) and was sub-cultured on TSA slant agar and incubated at 28°C for 24 hours. The obtained pure cultures were identified using the phenospecies and genospecies methods. The phenospecies characterization of A. hydrophila was performed in accordance with the Indonesian National Standard (SNI 7303, 2009), using Gram staining, motility test, oxidase test, blood agar test, oxidative-fermentative test and Rimler-Shoots+novobiocin test.

2.2 The isolation of

hydrophila ATCC 7699. While the amplicon resulted from the primer Cascón showed various results and sizes. DNA Amplification results using two pairs of primers showed different results from the same isolate. Figure 2 shows examples of various amplification results for the two tested pairs of primers. The seven isolates tested, i.e. A. hydrophila ATCC 7699, SfB, SfN, SfL, SfM, SfP, and PfKT9 had 685 bp amplicon; however, when amplified using the second primers, only the control isolate, SfB and PfKT9 had 760 bp amplicon. The isolate SfN had two amplifiers, which were different from the control isolate. Isolate SfL and SfP had single amplicon, but was different from the control, while the isolate SfM did not have