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ALIM ISNANSETYO
Department of Fisheries, Faculty of Agriculture, Universitas Gadjah Mada. Jl. Flora, Bulaksumur, Sleman 55281, Yogyakarta, Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology

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An antibacterial compound purified from a tropical coastal plant, Diospyros maritima ALIM ISNANSETYO; DESY PUTRI HANDAYANI; INDAH ISTIQOMAH; ADITYA ARIF; TAKUSHI KANEKO
Biodiversitas Journal of Biological Diversity Vol. 23 No. 1 (2022)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d230117

Abstract

Abstract. Isnansetyo A, Handayani DP, Istiqomah I, Arif A, Kaneko T. 2021. An antibacterial compound purified from a tropical coastal plant, Diospyros maritima. Biodiversitas 23: 135-142. This study aimed to isolate, purify, and characterize an antibacterial compound from Diospyros maritima and evaluate its antibacterial activity. Diospyros maritima bark was collected from Kei Kecil Island, extracted with ethanol, separated with hexane and chloroform, and purified by column chromatography. The isolated compound was tested for antibacterial activity by paper disk diffusion on the double-layer agar medium. The minimum inhibitory concentration (MIC) was determined by the microdilution method in a 96-well microplate. The pure compound inhibited Staphylococcus aureus and Aeromonas hydrophila at MICs of 0.625 and 5 µg/mL, respectively. The molecular weight of the compound was 188 m/z based on GC-MS analysis. The 1H-NMR spectrum indicated two protons which appeared as a multiplet at 7,687.59 ppm, one proton appeared as a singlet at 6.87 ppm and one proton appeared as a doublet at 7.26 ppm. The 13C-NMR spectrum showed eleven carbon signals, eight signals for the aromatic ring (186.06, 162.38, 151.17, 137.41, 134.49, 133.76, 17.75, and 16.51 ppm) and two signals at 191.91 and 191.05 ppm. The spectrometric data showed that the isolated compound was plumbagin (1,4-naphthalenedione-5-hydroxy-2-methyl). This finding opens up a new perspective for D. maritima as a source of lead compounds for the development of antibacterial substances.
The efficacy of Aeromonas hydrophila GPl-04 feed-based vaccine on African catfish (Clarias gariepinus) Dini Siswani Mulia; TRI UTOMO; ALIM ISNANSETYO
Biodiversitas Journal of Biological Diversity Vol. 23 No. 3 (2022)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d230339

Abstract

Abstract. Mulia DS, Utomo T, Isnansetyo A. 2022. The efficacy of Aeromonas hydrophila GPl-04 feed-based vaccine on African catfish (Clarias gariepinus). Biodiversitas 23: 1505-1510. Aeromonas hydrophila is a pathogenic bacterium to African catfish (Clarias gariepinus). Vaccination is one of the strategic efforts to control this type of bacterial attack. The objective of this study was to determine the effectiveness of A. hydrophila GPl-04 feed-based vaccine for African catfish. This study used A. hydrophila GPl-04 strain isolate as vaccine material. Sample used was African catfish measuring 10-13 cm length, weighing 16-25 g, taken from aquaculture ponds in the Purbalingga area, Central Java, Indonesia. This study applied experiments under the completely randomized design (CRD) method with 5 treatments and 3 replications. The treatments consisted of P0: feed without vaccine (control); P1: feed with 10 mL/100 g dose of vaccine given for 10 days; P2: feed with 10 mL/100 g dose of vaccine given for 15 days; P3: feed with 15 mL/ 100 g dose of vaccine given for 10 days; and P4: feed with 15 mL/ 100 g dose of vaccine given for 15 days. The challenge test was carried out on week 3, by injecting 0.1 mL of A. hydrophila suspension at 105CFU/mL dose per fish. The main parameters included the fish antibody titer, survival rate (SR), relative percent survival (RPS), mean time to death (MTD), and growth rate. Supporting parameters included water quality such as water temperature, water pH, and dissolved O2 levels. The main parameter data were analyzed using analysis of variance (ANOVA) and Duncan multiple range test (DMRT) at 5% test level, and the supporting parameter was descriptively quantitative analyzed. The results showed A. hydrophila GPl-04 feed-based vaccine could increase antibody titer, SR, RPS, and MTD of African catfish (P<0.05). In addition, vaccination did not adversely affect the growth. The A. hydrophila GPl-04 feed-based vaccine was effective in protecting African catfish from A. hydrophila attack where 10 mL/100 g dose, given for 10 days (P1) was indicated as the most effective compared to other vaccination treatments.
Antibiotic resistance of emerging pathogenic bacteria of hybrid grouper farming in Indonesia INDAH ISTIQOMAH; ALIM ISNANSETYO; MURWANTOKO MURWANTOKO; DESY PUTRI HANDAYANI; YANI NUR’AINI LESTARI; ARIEF TASLIHAN; I GUSTI NGURAH PERMANA; ENDANG WIJAYANTI
Biodiversitas Journal of Biological Diversity Vol. 24 No. 4 (2023)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d240465

Abstract

Abstract. Istiqomah I, Isnansetyo A, Murwantoko, Handayani DP, Lestari YN, Taslihan A, Permana IGN, Wijayanti E. 2023. Antibiotic resistance of emerging pathogenic bacteria of hybrid grouper farming in Indonesia. Biodiversitas 24: 2493-2501. A hybrid grouper is among the main coastal farming species in Asian countries, yet vibriosis is still a significant issue. Therefore, a knowledge of pathogenic strains responsible for recent cases is essential for developing appropriate control measures. The purpose of the study was to describe emerging bacterial pathogens associated with hemorrhagic disorder, skin wounds, and mortality in cantang hybrid grouper farms in Indonesia. A total of 30 bacterial strains were recovered from the kidney of diseased grouper on TCBS agar. Based on the ability to induce clinical signs and mortality of grouper juveniles owing to intraperitoneal bacterial infection at 106 CFU/fish, the bacteria were divided into three groups: pathogenic (n=11), low-pathogenic (n=2), and non-pathogenic (n=17). The pathogenic strains were putatively identified as Photobacterium damselae subsp. Damselae (n=8), Vibrio alginolyticus (n=1), Vibrio harveyi (n=2), and Vibrio azureus (n=2) based on the biochemical characteristics and 16S rRNA and DNA gyrase B subunit gene sequences. The pathogenic bacteria were sensitive to erythromycin, gentamycin, oxytetracycline, and kanamycin. However, it still has various resistant patterns to ampicillin, fosfomycin, chloramphenicol, streptomycin, rifampicin, and enrofloxacin. This study demonstrated the emergence of pathogenic P. damselae subsp. damselae, Vibrio alginolyticus, V. harveyi, and V. azureus in grouper aquaculture in Indonesia with multiple antibiotic resistance, which needed to further studies.
Antibacterial activity of mangrove plant extract of Rhizophora apiculata in inhibiting the growth of various strains of Aeromonas hydrophila DINI SISWANI MULIA; SUCI DWI RAHAYU; AMAN SUYADI; IKHSAN MUJAHID; ALIM ISNANSETYO
Biodiversitas Journal of Biological Diversity Vol. 24 No. 9 (2023)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d240921

Abstract

Abstract. Mulia DS, Rahayu SD, Suyadi A, Mujahid I, Isnansetyo A. 2023. Antibacterial activity of mangrove plant extract of Rhizophora apiculata in inhibiting the growth of various strains of Aeromonas hydrophila. Biodiversitas 24: 4803-4810. Aeromoniasis is a bacterial disease caused by Aeromonas hydrophila. Synthetic antibiotics are often used to treat this disease. However, using antibiotics for too long with excessive doses can cause side effects for the environment, fish, and other aquatic biota. One of the natural ingredients with the potential as an antimicrobial is the mangrove plant Rhizophora apiculata. This study aims to determine the antimicrobial activity of mangrove plant extract R. apiculata in inhibiting the growth of various strains of A. hydrophila. The study used a completely randomized design experimental method with a factorial pattern with 3 factors (plant part, bacterial strain, and extract concentration) with 48 treatments and 3 replications. Methanol (polar) and n-hexane (non-polar) were used as extract solvents. The parts of the plant used are leaves and stems. The bacterial strains used were GPl-04, GB-01, GK-01, GJ-01, GL-01, and GL-02. The concentration of mangrove extract used was 0, 10, 20, and 30%. The inhibition zone diameter measured the antimicrobial activity using a paper disc (Kirby Bauer method). The data analysis used was the non-parametric Kruskall-Wallis test and qualitative descriptive. The results showed that R. apiculata methanol extract's leaves and stems contain bioactive compounds of flavonoids, alkaloids, and tannins, while the n-hexane extract contains terpenoids. Leaves methanol extract R. apiculata exhibited better than stems, namely the inhibition zone diameter of 2.87 mm, at a concentration of 20% against the GJ-01 bacterial strain, while the methanol extract of stem R. apiculata showed the best antimicrobial activity, namely the diameter of the inhibition zone of 2.10 mm, at a concentration of 30% against the bacterial strain GL-02. Leaves n-hexane extract R. apiculata had the best antimicrobial activity at a concentration of 10% against the bacterial strain GL-01, with an inhibition zone diameter of 2 mm. The n-hexane extract of stems R. apiculata showed antimicrobial activity at a 30% concentration of 1.23 mm in the same strain. Mangroves plant R. apiculata have bioactive compounds that can be used as natural bactericides to control A. hydrophila bacterial disease.
Phenotypic and genotypic characteristics of pathogenic bacteria Aeromonas veronii bv veronii causes disease in gourami (Osphronemus gouramy) DINI SISWANI MULIA; ZAHROTUN NISA; SUWARSITO SUWARSITO; CAHYONO PURBOMARTONO; ALIM ISNANSETYO; NUNAK NAFIQOH; INA SALWANY MD YASIN; MOHAMAD AZZAM-SAYUTI
Biodiversitas Journal of Biological Diversity Vol. 25 No. 12 (2024)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d251248

Abstract

Abstract. Mulia DS, Nisa Z, Suwarsito, Purbomartono C, Isnansetyo A, Nafiqoh N, Yasin ISM, Azzam-Sayuti M. 2024. Phenotypic and genotypic characteristics of pathogenic bacteria Aeromonas veronii bv veronii causes disease in gourami (Osphronemus gouramy). Biodiversitas 25: 5103-5111. Gourami (Osphronemus gouramy) is a type of freshwater fish with excellent potential, but its cultivation is often disturbed by bacterial diseases caused by Aeromonas spp. The presence of virulent genes often influences the pathogenicity of Aeromonas spp. and treatment with antibiotics causes bacteria resistance. Therefore, this study aimed to determine the phenotypic and genotypic characteristics of Aeromonas veronii bv veronii causing disease in gourami (O. gouramy). A purposive sampling method was used to obtain specific diseased gourami. Phenotypic characteristics were determined morphologically and biochemically, while genotypic characteristics were based on 16S rDNA and virulent genes. A total of 10 virulent genes were used, including the aerA/haem, alt, ast, act, flaA, lafA, fstA, ahp, ela, and lip genes. Subsequently, antibiotic susceptibility tests were carried out using 10 µg gentamycin disk, 10 µg bacitracin, 30 µg tetracycline, and 30 µg chloramphenicol. Data on phenotypic and genotypic identification, the detection of virulence genes, and antimicrobial susceptibility tests were analyzed descriptively and qualitatively. The results showed that isolate BmSG-03 was closely related to A. veronii bv veronii strain ATCC 35624 with a similarity rate and query of 99.58% and 100%, respectively. The A. veronii bv veronii BmSG-03 isolate was detected to contain the aerA/haem, lafA, and ela genes, while the alt, ast, act, flaA, fstA, ahp, and lip genes were not detected. This isolate was resistant to bacitracin, tetracycline, and gentamycin but susceptible to chloramphenicol. Moreover, further investigations were recommended on genotypic characteristics of A. veronii bv veronii resistance gene and other antibiotic tests due to limited reports in Indonesia. For effective disease control, there should be a comprehensive database on the characteristics of the pathogen to obtain more effective and appropriate mitigation efforts.
Co-Authors ADITYA ARIF AMAN SUYADI ARIEF TASLIHAN CAHYONO PURBOMARTONO DESY PUTRI HANDAYANI DINI SISWANI MULIA DINI SISWANI MULIA Dini Siswani Mulia ENDANG WIJAYANTI I GUSTI NGURAH PERMANA IKHSAN MUJAHID INA SALWANY MD YASIN INDAH ISTIQOMAH MOHAMAD AZZAM-SAYUTI MURWANTOKO MURWANTOKO NUNAK NAFIQOH SUCI DWI RAHAYU SUWARSITO SUWARSITO TAKUSHI KANEKO TRI UTOMO YANI NUR’AINI LESTARI ZAHROTUN NISA