Achmad Dinoto
Research Center for Biology, Indonesian Institute of Sciences (LIPI). Jalan Raya Jakarta-Bogor KM 46, Cibinong 16911, Jawa Barat, Indonesia

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16s rRNA Identification of Pediococcus spp. from Broiler and Studies of Adherence Ability on Immobilized Mucus Damayanti, Ema; Yusiati, Lies Mira; Dinoto, Achmad
Indonesian Journal of Biotechnology Vol 17, No 2 (2012)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (309.077 KB)

Abstract

The objectives of this research were to study taxonomical status of lactic acid bacteria (LAB) isolated from broiler and adherence ability on mucus in vitro. Molecular analysis was performed by analyzing 16S rRNA gene using universal primer. The adherence assay on mucus was carried out using microplate method with total plate count (TPC), absorbance (A550) and confirmed by scanning electron microscopy (SEM). The results of this studies revealed that three of LAB isolates have closed relation to Pediococcus acidilactici (99.9%) species.Three isolates of P. acidilactici have adherence ability on broiler mucus higher than that on porcine mucin with an adherence percentage of 55.5% versus 50.8% and absorbance A550 of 0.061 versus 0.051, respectively. The highest adherence ability showed by P. acidilactici R02 with adherence percentage was 59.3% and absorbance A550 = 0.068. Adherence on mucus were affected by the addition of 3 g/l of gastric juice and 0.3% (b/v) of bile salt. Adherence analysis using SEM also showed that the adherence on broiler mucus was higher than the adherence on porcine mucin. Altogether this adherence studies, suggest that three isolates of P. acidilactici LAB were capable of colonizing host intestinal mucus in vitro as important property to be promising probiotic bacteria for broiler.Key words : adherence, broiler, Pediococcus, mucus, 16S rRNA
CHARACTERISTICS OF LACTIC ACID BACTERIA ISOLATED FROM GASTROINTESTINAL TRACT OF CEMANI CHICKEN AND THEIR POTENTIAL USE AS PROBIOTICS Jannah, Siti Nur; Dinoto, Achmad; Wiryawan, Komang Gede; Rusmana, Iman
Media Peternakan Vol. 37 No. 3 (2014): Media Peternakan
Publisher : Faculty of Animal Science, Bogor Agricultural University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (5594.722 KB) | DOI: 10.5398/medpet.2014.37.3.182

Abstract

The aims of this study were to screen and characterize lactic acid bacteria (LAB) isolated from gastrointestinal (GI) tract of Cemani chicken, one of Indonesian local chicken and to investigate their potential use as probiotics. LAB were isolated from GI tract using MRSA and GYPA media and incubated anaerobically. Selected LAB were determined their probiotic properties with several assays. Identification of selected LAB was based on 16S rDNA sequences, morphological and biochemical characteristics. Ninety five bacteria  were isolated and characterized as lactic acid bacteria (Gram positive, catalase negative, non sporeforming and acid producing). Twenty four isolates of LAB demonstrated antimicrobial activity against Escherichia coli JCM 1649 and Salmonella enteritidis B2586, and three selected isolates, i.e. CCM011, CSP004, and CVM002 showed the highest inhibition activity. The isolates had characters of high cell surface hydrophobicity and inter-isolate coaggregation ability of LAB, high survival at low pH, high  phytase and protease activity (but no amylase and lipase activity), weak coaggregation with pathogen and no resistance to the examined antibiotics. The isolates were identified based on sequence analysis of 16S rRNA gene as Lactobacillus salivarius, however, each isolate had different profiles of sugar fermentation. Therefore the three LAB isolates had potential application as probiotics for chicken.Key words: Cemani chicken, gastrointestinal tract, lactic acid bacteria, probiotic
16s rRNA Identification of Pediococcus spp. from Broiler and Studies of Adherence Ability on Immobilized Mucus Damayanti, Ema; Yusiati, Lies Mira; Dinoto, Achmad
Indonesian Journal of Biotechnology Vol 17, No 2 (2012)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (309.077 KB) | DOI: 10.22146/ijbiotech.7854

Abstract

The objectives of this research were to study taxonomical status of lactic acid bacteria (LAB) isolated from broiler and adherence ability on mucus in vitro. Molecular analysis was performed by analyzing 16S rRNA gene using universal primer. The adherence assay on mucus was carried out using microplate method with total plate count (TPC), absorbance (A550) and confirmed by scanning electron microscopy (SEM). The results of this studies revealed that three of LAB isolates have closed relation to Pediococcus acidilactici (99.9%) species.Three isolates of P. acidilactici have adherence ability on broiler mucus higher than that on porcine mucin with an adherence percentage of 55.5% versus 50.8% and absorbance A550 of 0.061 versus 0.051, respectively. The highest adherence ability showed by P. acidilactici R02 with adherence percentage was 59.3% and absorbance A550 = 0.068. Adherence on mucus were affected by the addition of 3 g/l of gastric juice and 0.3% (b/v) of bile salt. Adherence analysis using SEM also showed that the adherence on broiler mucus was higher than the adherence on porcine mucin. Altogether this adherence studies, suggest that three isolates of P. acidilactici LAB were capable of colonizing host intestinal mucus in vitro as important property to be promising probiotic bacteria for broiler.Key words : adherence, broiler, Pediococcus, mucus, 16S rRNA
Variasi Intraspesies Lactobacillus plantarum (Orla-Jensen) Bergey et al. Asal Sayur Asin Berdasarkan Analisis Molekuler ., Sulistiani; ., Abinawanto; Sukara, Endang; Dinoto, Achmad; Mangunwardoyo, Wibowo
JURNAL BIOLOGI INDONESIA Vol 11, No 1 (2015): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v11i1.2162

Abstract

The current study is the first report on intraspecies analysis of L. plantarum from sayur asin in Indonesia using molecular approach. Three molecular techniques, i.e., restriction fragment length polymorphism (RFLP) 16S-23S rDNA intergenic spacer region (ISR), random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and enterobacterial repetitive intergenic consensus (ERIC-PCR) were used to determine the intraspecies diversity of L. plantarum responsible for spontaneous fermentation in sayur asin. These methods were aimed to discriminate 46 isolates of L. plantarum isolated from sayur asin, including the type strain. PCR amplification of the 16S-23S rDNA ISR revealed two-bands profile of 800 and 600 bp specific to lactobacilli. RAPD-PCR and ERIC-PCR were very valuable in discriminating genetic polymorphism among L. plantarum isolates by producing bands ranged from 4-10 bands (360-2620 bp) and 6-12 bands (160-2900 bp), respectively. Dendograms generated from UPGMA cluster analysis based on RAPD-PCR and ERIC-PCR data showed that all isolates were grouped into three major clusters with 74% and 68.6% genetic similarity thresholds, respectively.The study indicated that strains belong to L. plantarum isolated from sayur asin were divided into three genotypic groups. Keywords: ERIC-PCR, Intraspecies, Lactobacillus plantarum, RAPD-PCR, RFLP 16S-23S rDNA ISR 
PERUBAHAN KADAR KOLESTEROL SERUM PADA TIKUS SETELAH MENGKONSUMSI MALTOOLIGOSAKARIDA YANG DISINTESIS SECARA ENZIMATIK MENGGUNAKAN AMILASE Bacillus licheniformis BLl [The Change of Serum Cholesterol Level in Rats after Consuming Maltooligosaccharide Synthesized by Enzimatic Reaction of Bacillus licheniformis BLl Amylase] Dinoto, Achmad; Rahayu, Rita Dwi; Satyaningtijas, Aryani S
BERITA BIOLOGI Vol 10, No 1 (2010)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1007.167 KB) | DOI: 10.14203/beritabiologi.v10i1.2046

Abstract

To investigate the impact of consuming enzymatically synthesized maltooligosaccharide using Bacillus licheniformis BLl amylase on reduction of serum cholesterol level, in preclinical study groups of Sprague Dawley rats were fed for 3 weeks with four different test diets: basal diet (BD), high serum cholesterol-stimulating basal diet (CD), high serum cholesterol-stimulating basal diet supplemented with 3% (w/w feed) commercial maltooligosaccharide (CI), and high serum cholesterol-stimulating basal diet supplemented with 3% (w/w feed) maltooligosaccharide synthesized by enzimatic reaction of B. licheniformis BLl amylase (CB).Our result shows a reduction of serum cholesterol level up to 39.1% in rats CB after three weeks consuming maltooligosaccharide synthesized using B. licheniformis BLl amylase. In addition, significantly increased blood glucose level was not found in this study as an impact of consuming maltooligosaccharide synthesized using B. licheniformis BLl amylase.
VARIASI GENETIK Lactobacillus fermentum Beijerink ASAL SAYUR ASIN BERDASARKAN ANALISIS RFLP 16S-23S rDNA ISR, RAPD-PCR DAN ERIC-PCR [Genetic Variation of Lactobacillus fermentum Beijerink Origin Sayur Asin Based on RFLP 16S-23S rDNA ISR, RAPD-PCR and ERIC-PCR Analysis] Sulistiani, Sulistiani; Mangunwardoyo, Wibowo; Abinawanto, Abinawanto; Sukara, Endang; Dinoto, Achmad; Salamah, Andi
BERITA BIOLOGI Vol 16, No 2 (2017)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3242.558 KB) | DOI: 10.14203/beritabiologi.v16i2.2772

Abstract

Molecular analysis of Lactobacillus fermentum isolates is essential to understand their genetic variation in relations to their roles in sayur asin fermentation process. Combination of three molecular techniques which is restriction fragment length polymorphism (RFLP) of 16S23S rDNA intergenic spacer region (ISR), random amplified polymorphic DNA (RAPD-PCR) and an enterobacterial repetitive intergenic consensus (ERIC-PCR) analysis were performed to discriminate 19 representative isolates of L. fermentum isolated from sayur asin. The result showed that L. fermentum strain D11 is distantly related to other isolates based on RFLP using HhaI restriction enzyme and RAPDPCR analyses. In addition, both of RAPD-PCR and ERIC-PCR successfully determined the genetic variation among L. fermentum strains by exhibiting distinct 4-8 bands (800-2080 bp) and 4-10 bands (280-3050 bp), respectively. A dendogram generated from UPGMA cluster analysis of both RAPD-PCR and ERIC-PCR data showed two distinct genotypic groups exist among L. fermentum isolated from sayur asin in Indonesia.
Pemisahan Eksopolisakarida (Eps) Sebagai Metabolit Bakteri Usus untuk Aditif Makanan dalam Biomassa Pati Sagu {Metroxylon sp.) dan Glukosa melalui Sistem Mikrofiltrasi Sel Berpengaduk (Separation of Exopolysaccharides (Eps) As Colon Bacteria Metabolismfor FoodAdditive in Sago Starch Biomass (Metroxylon sp.) and Glucose through Membrane Cell Microfiltration System) Susilowati, Agustine; Aspiyanto, Aspiyanto; Dinoto, Achmad; Lotulunga, Puspa D.
JURNAL PANGAN Vol 21, No 4 (2012): PANGAN
Publisher : Perum BULOG

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1224.818 KB) | DOI: 10.33964/jp.v21i4.203

Abstract

Kultur bakteri usus Lactobacillus sp. FU-0811 dan Enterobacter sp. FU-0813 yang ditumbuhkan pada medium berupa biomassa pati sagu (Metroxylon sp.) menghasilkan eksopolisakarida (EPS) yang berpotensi sebagai aditif makanan (pengental, pengemulsi, penstabil, pembawa). Penggunaan pati sagu merupakan alternatif biomassa selain glukosa. Melalui pemisahan dengan sistem membran mikrofiltrasi (MF) 0,15 pm berpengaduk diharapkan EPS dan metabolit lainnya diperoleh dengan konsentrasi lebih optimal. Penelitian ini bertujuan untuk mengetahui pengaruh pemisahan EPS hasil pertumbuhan bakteri usus Lactobacillus sp. FU-0811 dan Enterobacter sp. FU-0813 masing-masing dalam media biomassa pati sagu dan sebagai pembanding digunakan biomassa glukosa pada kondisi proses pemurnian tetap (kecepatan putar sel pengaduk 400 rpm, suhu ruang dan tekanan proses 40 psia) terhadap metabolit dengan komposisi terbaiksebagai bahan food aditif. Hasil penelitian menunjukkan bahwajenis biomassa, jenis mikroba dan sistem membran MF berpengaruh terhadap tingkat pemisahan metabolit. Retentat atau konsentrat hasil pemisahan memiliki komposisi dan jumlah mikroba lebih baik daripada permeat atau ekstrak untuk kedua jenis bahan tersebut. Proses membran MF terhadap biomassa glukosa dengan Lactobacillus sp. FU-0811 dan Enterobacter sp. FU-0813 masing-masing mampu menahan EPS sebagai gula reduksi dalam retentat atau konsentrat berturut-turut 73,73 persen dan 47,33 persen, serta pada biomassa pati sagu berturut-turut 95,5 persen dan 83,435 persen apabila dibandingkan dengan total gula reduksi dalam permeat dan retentat atau konsentrat pada masing-masing biomassa. Hasil idensifikasi metabolit melalui LC-MS memperlihatkan bahwa intensitas senyawa monosakarida dalam retentat atau konsentrat lebih tinggi daripada yang terdapat di dalam permeat.Colon bacteria culture of Lactobacillus sp. FU-0811 and Enterobacter sp. FU-0813 grown on biomass ofsago (Metroxylon sp.) produced exopolysaccharides (EPS) that have an important potential useas food additive (thickener, emulsifier, stabilizer and carrier). The useofsago starch isan alternative biomass beside glucose. By applying the stirred microfiltration membrane (0.15 pm ofpore size) cell, the biomass was separated to get EPS and other metabolites with more optimal concentration. The goal of this experiment was to find out separation effect ofEPS as a result of the growth ofcolon bacteria of Lactobacillus sp. FU-0811 and Enterobacter sp. FU-0813 in the biomass ofsagostarch. Asa comparison, glucose was used on fixed condition ofpurification process (rotation speed ofstirrer cell of400 rpm, room temperature and pressure of 40 psia) and the best composition of metabolite as food additive agent. The result showed that the type biomass, microbe, and MF membrane system influenced on the level of metabolite separation. The retentate or the concentrate of separation had better composition and microbial count than that of the permeate orextract for both biomasses. The process of MF membrane on glucose biomass with Lactobacillus sp. FU-0811 and Enterobacter sp. FU-0813 were subsequently able toretain EPSas reducing sugar in the retentate orconcentrate by 73.73 percent and 47.33percent, and the biomass ofsago starch by95.5percent and 83.435 percent when compared to total ofreducing sugar in permeate and retentate or concentrate for each biomass. The result ofmetabolite identification through LC-MS instrument displayed that greater intensity of monosaccharide compound was found in the retentate or concentrate than that of in the permeate. 
VARIASI GENETIK LACTOBACILLUS FERMENTUM BEIJERINK ASAL SAYUR ASIN BERDASARKAN ANALISIS RFLP 16S-23S RDNA ISR, RAPD-PCR DAN ERIC-PCR Sulistiani, Sulistiani; Mangunwardoyo, Wibowo; Abinawanto, Abinawanto; Sukara, Endang; Dinoto, Achmad; Salamah, Andi
BERITA BIOLOGI Vol 16, No 2 (2017)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v16i2.2772

Abstract

Molecular analysis of Lactobacillus fermentum isolates is essential to understand their genetic variation in relations to their roles in sayur asin fermentation process. Combination of three molecular techniques which is restriction fragment length polymorphism (RFLP) of 16S23S rDNA intergenic spacer region (ISR), random amplified polymorphic DNA (RAPD-PCR) and an enterobacterial repetitive intergenic consensus (ERIC-PCR) analysis were performed to discriminate 19 representative isolates of L. fermentum isolated from sayur asin. The result showed that L. fermentum strain D11 is distantly related to other isolates based on RFLP using HhaI restriction enzyme and RAPDPCR analyses. In addition, both of RAPD-PCR and ERIC-PCR successfully determined the genetic variation among L. fermentum strains by exhibiting distinct 4-8 bands (800-2080 bp) and 4-10 bands (280-3050 bp), respectively. A dendogram generated from UPGMA cluster analysis of both RAPD-PCR and ERIC-PCR data showed two distinct genotypic groups exist among L. fermentum isolated from sayur asin in Indonesia.
PERUBAHAN KADAR KOLESTEROL SERUM PADA TIKUS SETELAH MENGKONSUMSI MALTOOLIGOSAKARIDA YANG DISINTESIS SECARA ENZIMATIK MENGGUNAKAN AMILASE BACILLUS LICHENIFORMIS BLL Dinoto, Achmad; Rahayu, Rita Dwi; Satyaningtijas, Aryani S
BERITA BIOLOGI Vol 10, No 1 (2010)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v10i1.2046

Abstract

To investigate the impact of consuming enzymatically synthesized maltooligosaccharide using Bacillus licheniformis BLl amylase on reduction of serum cholesterol level, in preclinical study groups of Sprague Dawley rats were fed for 3 weeks with four different test diets: basal diet (BD), high serum cholesterol-stimulating basal diet (CD), high serum cholesterol-stimulating basal diet supplemented with 3% (w/w feed) commercial maltooligosaccharide (CI), and high serum cholesterol-stimulating basal diet supplemented with 3% (w/w feed) maltooligosaccharide synthesized by enzimatic reaction of B. licheniformis BLl amylase (CB).Our result shows a reduction of serum cholesterol level up to 39.1% in rats CB after three weeks consuming maltooligosaccharide synthesized using B. licheniformis BLl amylase. In addition, significantly increased blood glucose level was not found in this study as an impact of consuming maltooligosaccharide synthesized using B. licheniformis BLl amylase.
VARIASI INTRASPESIES LACTOBACILLUS PLANTARUM (ORLA-JENSEN) BERGEY ET AL. ASAL SAYUR ASIN BERDASARKAN ANALISIS MOLEKULER ., Sulistiani; ., Abinawanto; Sukara, Endang; Dinoto, Achmad; Mangunwardoyo, Wibowo
JURNAL BIOLOGI INDONESIA Vol 11, No 1 (2015): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v11i1.2162

Abstract

The current study is the first report on intraspecies analysis of L. plantarum from sayur asin in Indonesia using molecular approach. Three molecular techniques, i.e., restriction fragment length polymorphism (RFLP) 16S-23S rDNA intergenic spacer region (ISR), random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and enterobacterial repetitive intergenic consensus (ERIC-PCR) were used to determine the intraspecies diversity of L. plantarum responsible for spontaneous fermentation in sayur asin. These methods were aimed to discriminate 46 isolates of L. plantarum isolated from sayur asin, including the type strain. PCR amplification of the 16S-23S rDNA ISR revealed two-bands profile of 800 and 600 bp specific to lactobacilli. RAPD-PCR and ERIC-PCR were very valuable in discriminating genetic polymorphism among L. plantarum isolates by producing bands ranged from 4-10 bands (360-2620 bp) and 6-12 bands (160-2900 bp), respectively. Dendograms generated from UPGMA cluster analysis based on RAPD-PCR and ERIC-PCR data showed that all isolates were grouped into three major clusters with 74% and 68.6% genetic similarity thresholds, respectively.The study indicated that strains belong to L. plantarum isolated from sayur asin were divided into three genotypic groups. Keywords: ERIC-PCR, Intraspecies, Lactobacillus plantarum, RAPD-PCR, RFLP 16S-23S rDNA ISR