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Transposition and expression of GEP gene in the genome of Vibrio harveyi to monitor its adherence in shrimp larvae Antonius Suwanto
BIOTROPIA Vol. 13 No. 1 (2006)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (534.675 KB) | DOI: 10.11598/btb.2006.13.1.74

Abstract

Expression of green fluorescent protein encoded by GFP gene in Vibrio harveyi was investigated to understand the ability of the gene as a molecular marker for adherence of this pathogenic Vibrio in shrimp larvae. The GFP gene was inserted into pUC18Not and pUTmini-Tn5 to generate a recombinant plasmid pWGO2 and pWGO3, respectively, which was transferred into the three isolates of V. harveyi employing diparental mating. Recombinant E. coli carrying pWGO2 and pWGO3 resulted in green-fluorescent colonies and cells due to the production of GFP. However, al1 of mini-Tn5, including mini-Tn5-gfp were not successful1y transferred to V. harveyi. Therefore, we used mini-Tn10 (pLOFKm-gfp) for inserting of gfp gene into V. harveyi genome. Although we could obtain relatively high (l0 pangkat -8) transconjugans employing Tn10, only one of Tnl0 derived isolate of V. harveyi G3 (G3-Tn1Ogfp) showed gfp expression and was further employed for adherence assay. Fluorescent 03-TnlOgfp cells could be observed inside the digestive tract of shrimp larvae and could be distinguished from vibrio that naturally exist in shrimp larvae.
GENETIC DIVERSITY OF AMPICILLIN-RESISTANT Vibrio ISOLATED FROM VARIOUS STAGES OF TIGER SHRIMP LARVAE DEVELOPMENT WIDANARNI WIDANARNI; ANTONIUS SUWANTO
BIOTROPIA - The Southeast Asian Journal of Tropical Biology No. 15 (2000)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (219.218 KB) | DOI: 10.11598/btb.2000.0.15.160

Abstract

This research was carried  out to  study  genetic  diversity of ampicillin-resistant Vibrio  from various  stages of tiger  shrimp larvae  (Penaeus Monodon) development from,Tambak Inti Rakyat hatchery, near Labuan, West Java, Indonesia. A total of 25 ampicillin-resistant Vibrio isolates were isolated using thiosulphate  citrate bile-salt  sucrose  agar (TCBS-Agar) and seawater  complete agar (SWC-Agar). Physiological and biochemical characterization showed that the isolates could be grouped into only two species, i.e. V. harveyi from the egg stage; and V. metschnikovii from larvae and post-larval stage (i.e nauplius, zoea, mysis, PLi, PL5, PL,0, and PL,5). These isolates were also present in their respective rearing water  of  each stage  and  some natural feed.  Schizotyping analysis employing restriction endonuclease Noll (5'-GC4GGCCGC) indicated that the isolates could be grouped into  at least  13 different  genotypes.  Therefore,  schizotyping  was  more discriminative than physiological characterization. This study showed that particular groups of Vibrio colonized all stages of shrimp larvae and demonstrated closed phylogenetic relationship. These groups of Vibrio might be  the dominant microbiota which could suppress the development of other Vibrio including the pathogenic Vibrio. Key words : Shrimp/ampicillin-resistant K/fcno/schizotyping
A NOVEL INTEGRON IN THE GENOME OF ESCHERICHIA COLI ISOLATED FROM INDONESIAN MONITOR LIZARD (VARANUS SPP). ESTI PUSPITASARI; ANTONIUS SUWANTO; AMARILA MALIK; WALTER ERDELEN
BIOTROPIA - The Southeast Asian Journal of Tropical Biology No. 16 (2001)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (97.942 KB) | DOI: 10.11598/btb.2001.0.16.163

Abstract

The genotype of antibiotic resistance  in natural isolates of Escherichia coli was determined  through integron detection and  characterization of the associated antibiotic  resistance. E. coli SG2 isolated from Varanus salvator  of Java demonstrated  resistance  to spectinomycin (50ng/ml)  and streptomycin (SOng/ml). Integron detection indicated  that eight isolates out of nine E. coli  isolates possessed a conserved segment of the integron. Amplification of  the inserted cassette of the integron in this SG2 isolate yielded a 1-kb DNA fragment. Sequence analyses indicated that this fragment was homologous with aad gene, which confirmed  the resistance  to spectinomycin/streptomycin. This is the first report on the presence of integron in the E. coli isolated from the environment. Key words: Integron / antibiotic resistance / Escherichia coli
CHARACTERIZATION OF THREE BENZOATE DEGRADING ANOXYGENIC PHOTOSYNTHETIC BACTERIA ISOLATED FROM THE ENVIRONMENT DWI SURYANTO; ANTONIUS SUWANTO; ANJA MERYANDINI
BIOTROPIA - The Southeast Asian Journal of Tropical Biology No. 17 (2001)
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Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (152.215 KB) | DOI: 10.11598/btb.2001.0.17.166

Abstract

Three  anoxygenic photosynthetic bacteria, DS-1, DS-4 and Cas-13, have been examinated  for  themorphological and physiological  properties. All strains were rod-shape cells with  a swollen terminal  endGram  negative, motile, non-halophilic, non-alkalophilic  and non-acidophilic,  and capable of utilizinbenzoate aerobically and photo-anaerobically. Sequence analysis of part of 16S rRNA genes showed that DS1 and Cas-13 were closely related to Rhodopseudomonas palustris Strain 7 with a similarity of 97%, whereaDS-4 may not be closely related to the former two strains with a similarity of 78% based on the constructephylogenic  tree. Spectral analysis indicated that the three  bacteria  had  bacteriochlorophyl  a  and normaspirilloxanthin series. Growth in medium enriched with vitamin and supplemented with benzoate as their sole C-sources wabetter than in medium without vitamin. Benzoate degradation in medium with vitamin was accelerated. Thability  to grow on benzoate without added vitamins indicated  that  the bacteria were able to synthesize  theown vitamins. Key words: anoxygenic photosynthetic bacteria/ benzoate degradation/ 16S rRNA gene.
CHARACTERIZATION OF XYLANASE FROM A XYLANOLYTIC- THERMOPHILIC BACTERIUM ISOLATED FROM GUNUNG PANCAR HOT SPRING, WEST JAVA ANTONIUS SUWANTO; DWI SURYANTO; ANJA MERYANDINI
BIOTROPIA - The Southeast Asian Journal of Tropical Biology No. 17 (2001)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2001.0.17.167

Abstract

A xylanolytic-thermophilic bacterium (IT-08) was isolated from Gunung Pancar Hot Spring after two days of enrichment in Modified Thermus Medium (MTM) supplemented with 0.5% oat spelt xylan. 16S-rRNA sequence analysis indicated that IT-08 resembles Bacillus thermoleovorans, a species of thermophilic bacteria. When grown on xylan containing media, IT-08 produces a thermoactive xylanase. Xylanase from IT-08 was active at temperatures between 40 and 100°C, at pH values between 4.0 and 9.0 with optimum values obtained at 80°C and pH 6.0, respectively. SDS-PAGE and zymogram analysis demonstrated that a crude xylanase complex of IT-08 comprised two active bands with molecular masses of 78 and 60 kDa. Keywords: xylanase / xylanolytic-thermophilic bacteria
ADHERENCE AND PATHOGENICITY ASSAY OF VIBRIO HARVEYI IN TIGER SHRIMP (PENAEUS MONODON) LARVAE FOR SCREENING BIOCONTROL AGENT ANTONIUS SUWANTO; YUSMINAH HALA; RIDWAN AFFANDI; MUHAMMAD ZAIRIN Jr.
BIOTROPIA - The Southeast Asian Journal of Tropical Biology No. 18 (2002)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2002.0.18.171

Abstract

Rifampicin-resistant marker was employed as a reporter to detect the adherence and colonization of V. harveyi  in shrimp larvae. Vibrio harveyi P1B  and YA32.2 were isolated from dead shrimp larvae in Besuki, Northern Coast of East Java, while V. harveyi HB3, was isolated from pristine sea water in Pacitan, Southern Coast of East Java. Vibrio metschnikovii used as biocontrol agent was isolated from healthy shrimp larvae in Serang, West Java. Spontaneous mutation was conducted to generate V. harveyi P1B, YA32.2 and HB3 resistant to rifampicin. These mutants exhibited similar survival ability to their parental (wild type) strains. Significant larval mortality was observed in shrimp larvae inoculated with YA32.2 than that of larvae inoculated with P1B. Larvae  inoculated with HB3 showed the lowest mortality. Bacterial cell count of Vibrio Rf*  in dead  larvae were 103-104 cells/larvae. Isolates of Vibrio metschnikovii Z and M as biocontrol candidates effectively reduced the growth and adherence ability of YA32.2 to shrimp larvae. Larval mortality in rearing water inoculated simultaneously with YA32.2 and V. metschnikovii was lower than the one inoculated with YA32.2 alone. Therefore, Vibrio metschnikovii Z or M could be developed as an effective probiotic or biocontrol agent for V. harveyi in shrimp hatcheries. Key words :   Biological control/Vibrio metschnikovii/shrimp \arvae/Penaeus mwu«fon/pathogenicity assay/Vibrio harveyi
HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS ANTONIUS SUWANTO; AMARILA MALIK; BUDI TJAHJONO; ROB HARLING
BIOTROPIA - The Southeast Asian Journal of Tropical Biology No. 20 (2003)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (405.968 KB) | DOI: 10.11598/btb.2003.0.20.182

Abstract

A transcriptional fusion for an Aeromonas caviae chitinase gene was constructed under the control of a constitutive promoter of the kanaraycin resistance gene (PKmR). The construct was inserted into a medium copy number broad host range plasmid vector to yield recombinant plasmid pAM340, which harbored transcriptional fusion PKmR- chi. Another transcriptional fusion, Ptac-chi, in a recombinant plasmid pAM630, was conducted as comparison. Triparental mating of E. coli  carrying the recombinant plasmids with Pseudomotws fluorescens  5100, a phyllosphere bacterium, was performed. Pseudomonas fluorescens  5100 exconjugants were examined for constitutive expression of chitinase employing a spectrophotometric assay; they showed stronger chitin degradation activity than Escherichia coli transformants. Using a fungal antagonism plate assay, this chitinolytic P. fluorescens, however, could not inhibit selected phytopathogenic fungi. Keywords:    Aeromonas   caviae/  chitinase   gene/transcriptional   fusion/PKm'V   Vtac-chilPseudomonas fluorescens
SOLATION AND CHARACTERIZATION OF A NOVEL BENZOATE- UTILIZING Serratia marcescens DWI SURYANTO; ANTONIUS SUWANTO
BIOTROPIA - The Southeast Asian Journal of Tropical Biology No. 21 (2003)
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Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (424.953 KB) | DOI: 10.11598/btb.2003.0.21.184

Abstract

A new benzoate-utilizing strain, Serratia marcescens DS-8, isolated from the environment was characterized. The strain was enterobacilli, Gram negative, mesophilic, non halophilic, and aerobic bacterium that showed motile ovale-rod shaped cells. The isolate produced extracellular chitinase, protease, and prodigiosin (a red pigment produced by several Serratia strains yielding bright red or pink colonies). A physiological assay using Microbact* test showed that the strain was closely related to Klebsiella ozaenae  (49.85%) and Serratia liquefaciens  (24.42%), respectively. However, 16S rRNA sequence analysis indicated that the strain was closely related to  S. marcescens  DSM 30121 with similarity level of 98%. DS-8 strain was able to synthesize its own vitamins. Optimum growth in benzoate was obtained at pH between 7-8.5 and NaCl concentration of 1-1.5% (w/v). The isolate could grow in benzoate-containing medium up to 10 mM. Other carbon sources that could support the growth of DS-8 were casamino acid, glutamate, glucose, acetate, potato starch, and ethanol. Keywords: Serratia marcescens/aromatic degradation/168 rRNA sequence
ISOLATION AND CHARACTERIZATION OF MANNANOLYTIC THERMOPHILIC BACTERIA FROM PALM OIL SHELL AND THEIR MANNANASE ENZYME PRODUCTION PROPERTIES ANTONIUS SUWANTO; SUMARDI; MAGGY T HENAWIDJAJA; T RESNAWATI P URWADARIA
BIOTROPIA - The Southeast Asian Journal of Tropical Biology No. 25 (2005)
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Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2005.0.25.211

Abstract

A mannanolytic thermophilic bacterium (L-07) was isolated from palm oil shell after 2 days ofenrichment in liquid medium supplemented with 1% palm kernel meal as mannan source. Sequenceanalysis of 16S-rRNA indicated that L-07 was similar (98%) to  Geobacillus stearothermophilus, aspecies of thermophilic aerobic bacteria. We found that  G. stearothermophilus L-07 producedextracellular β-1,4-mannanases, but no β-manosidase and α-galactosidase activities. The growth of L-07reached its maximum (3.0 x 106 cell/ml) at 12-20 hours, while the highest  β-mannanase activity (0.52U/ml) was observed in culture medium after 36 hours of cultivation at 60oC. The medium containinglocust bean gum was the best for producing extracellular β-1,4-mannanases compared with kolang kaling,konjak, and palm kernel meal. SDS-PAGE and zymogram analysis demonstrated that crude mannanasecomplex of L-07 from locust bean gum containing medium comprised three active bands with molecularweight of 85, 73 and 50 kDa.   Keywords :  Extracellular enzyme/mannanase/Geobacillus stearothermophilus
DYNAMICS OF MICROBIAL COMMUNITY DURING TEMPEH FERMENTATION Antonius Suwanto
BIOTROPIA - The Southeast Asian Journal of Tropical Biology Vol. 28 No. 1 (2021): BIOTROPIA Vol. 28 No. 1 April 2021
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (524.271 KB) | DOI: 10.11598/btb.0.0.0.820

Abstract

Tempeh is sliceable soybean-cake fermented by Rhizopus oligosporus. Various bacteria were detected in tempeh employing cultivation technique with limited information about their origin or sources. The present study aimed to examine the source/s of bacterial community in tempeh by combining metagenomics analysis and cultivation technique. Samples were obtained from a number of steps in tempeh production employing two-times boiling of soybean (WJB tempeh production). All samples were plated on Enterobacteriaceae and Lactic Acid Bacteria medium. Total DNA were extracted directly from tempeh for metagenomics analysis, employing High-Throughput Sequencing (HTS) and cloned 16S rRNA genes. Firmicutes and Proteobacteria were the predominant and second dominant bacteria existed in fresh tempeh (FT) obtained by metagenomics analysis. In contrast, cultivation technique showed that Proteobacteria was the predominant phylum, suggested that most of the Firmicutes were not culturable. FT was dominated by Lactobacillus and Acetobacter. Both FT and soaking water (SW) were dominated by same species of Lactobacillus, i.e. L. delbreuckii and L. mucosae, indicated that SW was probably the source of bacterial community established in the final product of fermentation. Predominant bacteria in starter culture (SC), Acinetobacter, was not detected in FT, indicating that bacteria in SC might not play significant role in bacterial community development in FT.
Co-Authors . EFRIWATI . Melani . NURHAIMI-HARIS A A Hermosaningtyas A A Hermosaningtyas, A A ADI YULANDI Agus PURWANTARA Agus Purwito Agustin Wydia Gunawan Agustina, Delia Alfred Michael ALINA AKHDIYA RUSMANA AMARILA MALIK AMARILA MALIK Andi Khaeruni Andi Khaeruni Andreas Adhi Satya Andreas Adhi Satya ANJA MERYANDINI Anja Meryandini ARI SUSILOWATI Arifudin Rafif , Muhammad Ghildan Arild Ranlym Arifin Arina Amalia Putri Aris Tjahjoleksono Aris Tri Wahyudi ARTINI PANGASTUTI Asmini Budiani Basil J NIKOLAU BIBIANA W LAY BUDI TJAHJONO BUDI TJAHJONO Budi Tjahjono Budi Tjahjono Budinarta, Widyah C Hanny Wijaya CAHYA PRIHATNA Cahya Prihatna Cahya Prihatna CECILIA ANNA SEUMAHU CECILIA ANNA SEUMAHU DEBORA HADISUSANTO Delia Agustina Desniar - - DIAH ISKANDRIATI DIANA ELIZABETH WATURANGI Djoko Santoso Dondin Sajuthi DWI SURYANTO Dyah Kusuma Anggraini Edi Husen Edi Husen EDI HUSEN ERNIN HIDAYATI Esti Puspitasari ESTI PUSPITASARI ESTI PUSPITASARI Esti Utarti Eunice Limantara EVELINE AYU Felicia Felicia Felicia Kartawidjajaputra Ferymon Mahulette Ferymon Mahulette Ferymon Mahulette, Ferymon G. A. Wattimena Gayuh Rahayu Griselda Griselda Griselda Herman Natadiputri Griselda, Griselda Hajrial ASWIDINNOO HAJRIAL ASWIDINNOOR Hariyatun Hariyatun Hariyatun, Hariyatun Hermosaningtyas, Anastasia Aliesa Iman Rusmana Inez H.S. Loeddin Suharsono It Jamilah IVAN HANJAYA JOANITA SADELI KATHARINA JESSICA Kusharyoto, Wien Lilis Nuraida Linda Wati LISTYA UTAMI KARMAWAN Ludovika Jessica Virginia MAGGY T HENAWIDJAJA Maggy T Suhartono Maggy T Suhartono MAGGY T. SUHARTONO Maggy T. Suhartono MAGGY T. SUHARTONO MAGGY THENAWIJAYA SUHARTONO Maria Dita Febriani Lumban Gaol Maria Indah Purnamasari Maria Indah Purnamasari Maria Indah Purnamasari Maria Sugiharti Meity S. Sinaga MEITY SURADJI SINAGA Muhamad Azwar Syah Muhammad Agus Muljanto MUHAMMAD ZAIRIN Jr. Ni Nyoman Tri Puspaningsih NISA RACHMANIA MUBARIK NURITA TORUAN-MATHIUS NURUL AINI PRIHASTO SETYANTO Prihasto Setyanto Prihatna , Cahya Puspitasari, Esti QURROTA A’YUN R. Sapto Hendri Boedi Soesatyo Raden Ajie Syahbarie RAHAYU WIDYASTUTI RASTI SARASWATI RASTI SARASWATI Rasti Saraswati Rasti Saraswati RATIH DEWI HASTUTI RATNA SETYANINGSIH RATNA SETYANINGSIH RIDWAN AFFANDI Rika Indri Astuti ROB HARLING ROHANI CINTA BADIA GINTING Rustam, Yepy Hardi Satya, Andreas Adhi Sheila Sutanto Sheila Sutanto, Sheila SUMARDI Suryo Wiyono SUSAN SOKA SUSILOWATI1 SUSILOWATI1 T RESNAWATI P URWADARIA Tan Watumesa Agustina TARUNI SRI PRAWAST MIEN KAOMINI ANY ARYANI DEDY DURYADI SOLIHIN Tati Barus TEMMY DESILIYARNI TRESNAWATI PURWADARIA VICKY MEICY WALTER ERDELEN WIDANARNI WIDANARNI Widanarni Widanarni Wien Kusharyoto Yasinta Ratna Esti Wulandari Yepy Hardi Rustam Yogiara Yogiara YULIN LESTARI Yusminah Hala