Indonesian Journal of Biotechnology
The Indonesian Journal of Biotechnology (IJBiotech) is an open access, peer-reviewed, multidisciplinary journal dedicated to the publication of novel research in all aspects of biotechnology, with particular attention paid to the exploration and development of natural products derived from tropical—and especially Indonesian—biodiversity. IJBiotech is published biannually and accepts original research articles featuring well-designed studies with clearly analyzed and logically interpreted results. A strong preference is given to research that has the potential to make significant contributions to both the field of biotechnology and society in general.
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Effect of Probiotic Lactobacillus sp. Dad13 on Humoral Immune Response of Balb/C Mice Infected with Salmonella typhimurium
Kusumawati, Ika Dyah;
Harmayani, Eni;
Asmara, Widya
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
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An indigenous strain of lactic acid bacterium (LAB) identified as Lactobacillus spp. Dad13 (Dad13), isolatedfrom traditional fermented buffalo milk, was found to be potential as probiotic. The aim of this research was to studythe effect of probiotic Dad13 on humoral immune response of Balb/C mice infected with Salmonella typhimurium. Thespecific objective was to find out the effect of different Dad13 consumption time (before and along with infection of S.typhimurium) on the humoral immune response of Balb/C mice. The experiment was conducted by in vivo trial on 20males of Balb/C mice, age of 6-8 weeks, fed with AIN-93 standard diet. The mice were assigned into 4 groups. Eachgroup received the following treatments, ie. Dad13 only, Dad13 before infection, Dad13 along with infection andSalmonella infection only. A volume of 100 μl Dad13 cell suspensions (1010 CFU/ml) were given by oral forced feedingdaily for a week, at week 3 for group before infection and at week 4 for group of Dad13 only and Dad13 along withinfection. Salmonella infection (109 CFU/ml) was given once orally at week 4 to all groups except group treated withDad13 only. The humoral immune response of Balb/C mice was detected 2 weeks after infection by measuring thetiters of IgG and IgA specific from serum and mucosal intestinal liquid samples using Enzyme-linked ImmunosorbentAssay (ELISA) method. The result indicated that humoral immune response of Balb/C mice consuming Dad13 beforeand along with Salmonella infection were significantly different (p<0.05). Dad13 consumption along with Salmonellainfection increased circulated IgG and IgA as well as secretory IgA. It can be concluded that Dad13 probiotic feedingalong with infection increased humoral immune response more significantly compared to that before infection.Key words : Probiotic, Lactobacillus sp. Dad13, Immune response, Salmonella typhimurium
In vitro Antiplasmodial Activity and Cytotoxicity of Vincadifformine and Its Semisynthetic Derivatives
M, Mustofa;
Mallié, Michèle;
Valentin, Alexis;
Lewin, Guy
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
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An indole alkaloid with aspidospemane structure possessing a potential antiplasmodial activity,vincadifformine, has been isolated from Aspidosperma pyrifolium Mart. Moreover, 10 derivatives were preparedfrom the vincadifformine. The study was conducted to evaluate the in vitro antiplasmodial and cytotoxic activity ofthe vincadifformine and their semisynthetic derivatives. The in vitro antiplasmodial activity was evaluated onPlasmodium falciparum chloroquine-resistant (FcM ) and –sensitive (Nigerian) strains after 24-h and 72-h incubation, 29while cytotoxic activity was estimated on Hela cells and Cytotoxicity Index (CI = IC on HeLa cells/IC on FcM strain) 50 50 29was calculated to evaluate the safety of tested compounds. Experiment results showed that two compounds (4 and 8)exhibited good antiplasmodial activities in comparison with parent compound, vincadifformine and other testedcompounds with IC ranging from 5.3 to 12.8 μM on FcM strain and 11.4 to 24.0 μM on Nigerian strain. In addition, 50 29the CI of two compounds were also lower after 24-h incubation (CI, 2.0 and 4.8) than that of after 72-h incubation (CI,9.5 and 11.5). Further study will be conducted to evaluate quantitative structure-activity relationship (QSAR) in orderto design new antimalarial drugs.Keywords : vincadifformine - antiplasmodial – Plasmodium falciparum – cytotoxic - HeLa
Detection of eae, bfpA, espA Genes on Diarrhoeagenic Strains of Escherichia coli Isolates
Harti, Agnes Sri;
Iravati, Susi;
Asmara, Widya
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
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The Enteropathogenic Escherichia coli (EPEC) is one of pathogenic strain of diarrheagenic E. coli group in children andinfant that occurs in developing countries. The significant virulence factors in pathogenic EPEC are eaeA (E. coli attachingeffacing), bfpA (bundle-forming pilus A) and espA (encoding secreted protein A) genes. The use of DNA probes to detect thevirulence genes in E. coli in Indonesia is not common yet. In this experiment the gene fragments of eae, bfpA, and espA were usedas probes to detect the EPEC among E. coli isolates from stool specimensin of diarrheic children attending Public Health Centersin Yogyakarta. The DNA samples were isolated from 49 diarrheagenic E. coli isolates. The DNA probes of eae, bfpA and espAwere obtained by amplification of DNA fragment of EPEC O126 using PCR technique. Furthermore, those probes were used toidentify the presence of those genes among E. coli isolates using hybridization technique. The results showed that 42 (85.7%)isolates were espA+, 25 isolates (51%) were eaeA+ (EPEC strains). Therefore among 25 isolates of EPEC, 20 isolates (80 %)among EPEC were bfpA+ (typical EPEC strains).Keywords : DNA probe, eae, bfpA, espA, EPEC.
Production of Poly-ñ-hydroxybutyrate (PHB) from Sago Starch by The Native Isolate Bacillus megaterium PSA10
Yanti, Nur Arfa;
Sembiring, Langkah;
Margino, Sebastian
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
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A new bacterial strain that produces amylase and poly-α-hidroxybutyrate (PHB) using sago starch as carbon source was characterized and identified to be member of the Bacillus megaterium group based on phenotypic characteristics and 16S rDNA gene sequences. Amylase activity was determined spectrophotometrically on the basis of substrate concentration reduction. PHB production was quantified with UV spectrophotometer. The polymer produced by B. megaterium PSA10 was identified by Fourier Transform Infrared spectroscopy (FTIR). The result of the study showed that the amylase specific activity B. megaterium PSA10 was 593,61 DUN/mg protein and PHB production from sago starch was 52,28 % of cell dry weight (CDW). FTIR analysis of the polymer indicated that the strain B.megaterium PSA10 was a potent PHB producer.
Apoptosis and Phagocytosis Activity of Macrophages Infected by Mycobacterium tuberculosis Resistant and Sensitive Isoniazid Clinical Isolates
Rachmawaty, Farida J.;
Wibawa, Tri;
Soesatyo, Marsetyawan H.N.E.
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
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Mycobacterium tuberculosis (M.tb) is the main causative pathogen that cause the pulmonary tuberculosis.Intracellular M.tb was reported able to induce macrophages apoptosis, which may have crucial role in the regulationof immun response against M.tb infection. As an intracellular bacteria, M.tb able to live and replicate withinmacrophages. Phagocytosis is the first step to achieved this condition. The induction of macrophages apoptosis byINH resistant and sensitive M.tb clinical isolates, and H37Rv was studied. The macrophages apoptosis level weremeasured using an Ag-capture ELISA for histone and fragmented DNA (Cell Death Detection ELISAplus, RocheDiagnostic GmBH). Phagocytosis activity also analyzed, after staining using fluorescence dye (AcriFluorTM, ScientificDevice Lab.). The results showed that there was no significantly different between INH resistant and sensitive M.tbclinical isolates in respect their ability to induce apoptosis. The phagocytosis activity among the clinical isolates wasshown to be strain dependent, and undistinguishable between the Mtb clinical isolates. There was no associationbetween macrophages apoptosis level and the phagocytosis activity. These data suggested that among the virulentMtb clinical isolates, the ability to induce macrophages apoptosis and phagocytosis were consistently in comparablelevelKeywords: Mycobacterium tuberculosis, apoptosis, phagocytosis, macrophages, isoniazid
Developmental Competence of Early Stage Porcine Embryos Cultured in Medium with Different Energy Substrate in vitro
Karja, Ni Wayan Kurniani;
Kikuchi, Kazuhiro;
Fahrudin, Mokhamad
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
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To elucidate the effect of energy requirement during the early embryonic development on their developmentalability to develop to blastocyst stage, in vitro fertilized (IVF) porcine one-cell embryos were cultured in modifiedNorth Carolina State University (NCSU)-37 supplemented with different energy substrate. Result indicated that thecleavage rate of embryos in Pyr-Lac and Gluc-Pyr-Lact groups was significantly higher than in those in Gluc groupand Gluc-Rib group (P < 0.05). At Day 6 of culture, the highest proportion of embryos develop to the blastocyst stagewas obtained in the presence of pyruvate-lactate only. In the medium with glucose, the addition of pyruvate-lactateor ribose slightly increased the proportion of embryos develop to the blastocyst stage, however the value were notsignificantly different form those obtained in the presence of glucose only. The mean cell number in blastocystsderived from Pyr-Lac and Gluc-Pyr-Lact groups were significantly higher than those in the Gluc group (P < 0.05).These results indicated that the presence of glucose only, as energy substrate, during the first 2 days of in vitro culture(IVC) caused a decrease in development of in vitro produced (IVP) porcine embryos to the blastocyst stage and meancell number in blastocysts .Keywords: porcine embryos-energy substrate-in vitro culture
In vitro Antiplasmodial Activity and Cytotoxicity of Vincadifformine and Its Semisynthetic Derivatives
M. Mustofa;
Michèle Mallié;
Alexis Valentin;
Guy Lewin
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
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DOI: 10.22146/ijbiotech.7562
An indole alkaloid with aspidospemane structure possessing a potential antiplasmodial activity, vincadifformine, has been isolated from Aspidosperma pyrifolium Mart. Moreover, 10 derivatives were prepared from the vincadifformine. The study was conducted to evaluate the in vitro antiplasmodial and cytotoxic activity of the vincadifformine and their semisynthetic derivatives. The in vitro antiplasmodial activity was evaluated on Plasmodium falciparum chloroquine-resistant (FcM29) and –sensitive (Nigerian) strains after 24-h and 72-h incubation, while cytotoxic activity was estimated on Hela cells and Cytotoxicity Index (CI = IC50 on HeLa cells/IC50 on FcM29 strain) was calculated to evaluate the safety of tested compounds. Experiment results showed that two compounds (4 and 8) exhibited good antiplasmodial activities in comparison with parent compound, vincadifformine and other tested compounds with IC50 ranging from 5.3 to 12.8 µM on FcM29 strain and 11.4 to 24.0 µM on Nigerian strain. In addition, the CI of two compounds were also lower after 24-h incubation (CI, 2.0 and 4.8) than that of after 72-h incubation (CI, 9.5 and 11.5). Further study will be conducted to evaluate quantitative structure-activity relationship (QSAR) in order to design new antimalarial drugs.
Detection of eae, bfpA, espA Genes on Diarrhoeagenic Strains of Escherichia coli Isolates
Agnes Sri Harti;
Susi Iravati;
Widya Asmara
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
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DOI: 10.22146/ijbiotech.7563
The Enteropathogenic Escherichia coli (EPEC) is one of pathogenic strain of diarrheagenic E. coli group in children and infant that occurs in developing countries. The significant virulence factors in pathogenic EPEC are eaeA (E. coli attaching- effacing ), bfpA (bundle-forming pilus A) and espA (encoding secreted protein A) genes. The use of DNA probes to detect the virulence genes in E. coli in Indonesia is not common yet. In this experiment the gene fragments of eae, bfpA, and espA were used as probes to detect the EPEC among E. coli isolates from stool specimensin of diarrheic children attending Public Health Centers in Yogyakarta. The DNA samples were isolated from 49 diarrheagenic E. coli isolates. The DNA probes of eae, bfpA and espA were obtained by amplification of DNA fragment of EPEC O126 using PCR technique. Furthermore, those probes were used to identify the presence of those genes among E. coli isolates using hybridization technique. The results showed that 42 (85.7%) isolates were espA+, 25 isolates (51%) were eaeA+ (EPEC strains). Therefore among 25 isolates of EPEC, 20 isolates (80 %) among EPEC were bfpA+ (typical EPEC strains).
Production of Poly-α-hydroxybutyrate (PHB) from Sago Starch by The Native Isolate Bacillus megaterium PSA10
Nur Arfa Yanti;
Langkah Sembiring;
Sebastian Margino
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
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DOI: 10.22146/ijbiotech.7559
A new bacterial strain that produces amylase and poly-α-hidroxybutyrate (PHB) using sago starch as carbon source was characterized and identified to be member of the Bacillus megaterium group based on phenotypic characteristics and 16S rDNA gene sequences. Amylase activity was determined spectrophotometrically on the basis of substrate concentration reduction. PHB production was quantified with UV spectrophotometer. The polymer produced by B. megaterium PSA10 was identified by Fourier Transform Infrared spectroscopy (FTIR). The result of the study showed that the amylase specific activity B. megaterium PSA10 was 593,61 DUN/mg protein and PHB production from sago starch was 52,28 % of cell dry weight (CDW). FTIR analysis of the polymer indicated that the strain B.megaterium PSA10 was a potent PHB producer.
Apoptosis and Phagocytosis Activity of Macrophages Infected by Mycobacterium tuberculosis Resistant and Sensitive Isoniazid Clinical Isolates
Rachmawaty, Farida J.;
Wibawa, Tri;
Soesatyo, Marsetyawan H.N.E.
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada
Show Abstract
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Full PDF (43.035 KB)
Mycobacterium tuberculosis (M.tb) is the main causative pathogen that cause the pulmonary tuberculosis.Intracellular M.tb was reported able to induce macrophages apoptosis, which may have crucial role in the regulationof immun response against M.tb infection. As an intracellular bacteria, M.tb able to live and replicate withinmacrophages. Phagocytosis is the first step to achieved this condition. The induction of macrophages apoptosis byINH resistant and sensitive M.tb clinical isolates, and H37Rv was studied. The macrophages apoptosis level weremeasured using an Ag-capture ELISA for histone and fragmented DNA (Cell Death Detection ELISAplus, RocheDiagnostic GmBH). Phagocytosis activity also analyzed, after staining using fluorescence dye (AcriFluorTM, ScientificDevice Lab.). The results showed that there was no significantly different between INH resistant and sensitive M.tbclinical isolates in respect their ability to induce apoptosis. The phagocytosis activity among the clinical isolates wasshown to be strain dependent, and undistinguishable between the Mtb clinical isolates. There was no associationbetween macrophages apoptosis level and the phagocytosis activity. These data suggested that among the virulentMtb clinical isolates, the ability to induce macrophages apoptosis and phagocytosis were consistently in comparablelevelKeywords: Mycobacterium tuberculosis, apoptosis, phagocytosis, macrophages, isoniazid