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Articles by issue : Vol 4, No 1 (2010): April 2010
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Genetic Diversity of Yeasts from Fermented Orange Juice Based on PCR-RFLP and Sequence Analysis of the Internal Transcribed Spacer Regions

SOKA, SUSAN ( Universitas Katolik Indonesia Atma Jaya ) , SUSANTO, ANASTASIA ( Universitas Katolik Indonesia Atma Jaya )

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

Orange is one of the most valuable and common fruits in Indonesia. High glucose level in orange juice provides good growth conditions for yeasts. In this study, yeasts were isolated from fermented orange juice and subjected to diversity analysis. The analysis was conducted using restriction fragment length polymorphism on the internal transcribed spacer (ITS) region (including ITS1, 5.8S rRNA gene and ITS2), which was amplified using PCR with ITS1 and ITS4 primers.Restriction enzymes used in this research were HhaI, HinfI and HaeIII. A total of 24 yeast isolates were obtained from three different kinds of fermented orange juices (Indonesian Medan orange, Sunkist orange and Indonesian Pontianak orange). RFLP analysis of ITS regions revealed different amplified PCR fragment sizes and restriction profiles for each type of orange juice. However, all yeasts isolated from the same type of orange juice showed identical restriction patterns. Sequencing of ITS regions showed that three different yeast species were detected from each type of orange, e.g. Pichia veronae from Indonesian Pontianak orange, Cryptococcus albidosimilis from Sunkist orange and Issatchenkia orientalis from Indonesian Medan orange.

The Production of Nata Colored by Monascus purpureus J1 Pigments as Functional Food

PURWADARIA, TRESNAWATI, GUNAWAN, LISRINA, GUNAWAN, AGUSTIN WYDIA

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

Pigments from Monascus sp. may color nata. The mold also produces monacolin K that inhibits HMG-CoA reductase affecting the reduction of blood cholesterol. The aim of this study was to produce the colored nata containing monacolin K. The mold was isolated from commercial angkak. Potato sucrose (PS) and synthetic glucose (SG) media were used to ferment nata with Monascus purpureus J1. Fermentation of nata in PS medium produced red nata, while that in SG medium produced orange nata. The color of nata was similar to the color of supernatant. The optimum red production was obtained after five days of incubation, while the orange production increased until the 14th day. The color concentrations of the supernatant of PS medium containing nata (35.4 μg mL-1) were lower than those without nata (12.4 μg mL-1). The colors of nata looked darker than the color of the supernatant. The concentration of monacolin K in the red nata and the supernatant of PS medium were 0.6 μg mL-1 and 4.6 μg mL-1 respectively, while those in the orange nata and the supernatant of  SG  medium were 3.2 μg mL-1 and 14.6  μg mL-1 respectively. Dry matter biomass in the PS medium was lower than that in the SG medium. Even though the color of nata looked relatively stable, analyses of the nata water extract  that showed a stable condition only occurred in  freezing (-20OC) and soaking in buffer solution of pH 12; boiling, water  washing, and soaking in a solution of pH 3 and 7 reduced the pigment concentration. Monacolin K concentration was not stable for every treatment, especially for water washing and freezing. Eventhough it was not stable, the boiling nata contained red pigments and monacolin K of 19.7 μg mL-1 and 0.1 μg mL-1 respectively, which can be served as functional food.

Amantadine Resistant of Indonesian H5N1 Subtype Influenza Viruses During 2003-2008

DHARMAYANTI, NI LUH PUTU INDI, IBRAHIM, FERA, SOEBANDRIO, AMIN

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

The M2 protein of 146 avian influenza (AI) viruses data available in public database (NCBI), including 20 AI isolates used in this study, were sequenced at the M2 protein to find out the probability of mutation and the increase of resistance to amantadine after more than 5 years of their circulation in Indonesia. The results showed that during 2003-2008, around 62.58% (92/147) AI viruses in Indonesia have showed resistance to amantadine and 10 of them have dual mutations at V27A and S31N.

Effect of Oxygenated Water and Probiotic Administration on Fecal Microbiota of Rats

SURONO, INGRID SURYANTI, KHOMSAN, ALI, SOBARIAH, ENOK, NURANI, DARTI

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

Oxygenated water is  water with increased concentration of physically dissolved oxygen, and can perform all the same functions as the oxygen absorbed through the lungs. Several structures of human organs participate in the absorption and transportation of the oxygen, including the villi and cells containing mitochondrion in the small intestine as well as the lymph system.  The aim of this in vivo study were three folds, to validate the support of oxygenated water on viability of probiotic bacteria in the GUT, to suppress the fecal coliform, and to study the effect of oxygen concentration on the profile of fecal microbiota. There were one control group and three probiotic groups of 5 rats each based on strain of probiotic supplementation, control without probiotic (a0), Lactobacillus casei commercial strain (a1), Lactobacillus sp. IS-7257 (a2) and Lactobacillus sp. IS-27560 (a3). Each group was treated with three variable treatments, without oxygenated water supplementation (b0),  supplemented with oxygenated water at  50 ppm (b1), and  at  80 ppm (b2). Fecal samples were collected before (c0), after 3 days (c1), 7 days (c2)   supplementation,  followed by 3 days after returning back to normal diet (c3), analysed by culture dependent analyses for viable fecal lactic, coliform and fecal anaerobic bacteria. Supplementation of oxygenated water at 50 ppm, significantly increase fecal lactic acid bacteria of all  probiotic groups after 3 and 7 days (P<0.05);  80 ppm oxygenated water tends to lower the fecal coliform (P<0.1), while oxygenated water administration gives no effect on fecal anaerobic bacteria. As a conclusion, 50 ppm oxygenated water administration significantly  increased viable fecal lactic acid bacteria in probiotic groups. On the other hand, 80 ppm oxygenated water administration  tends to lower the fecal coliform bacteria. No effect of administration probiotic and/or oxygenated water on viability of fecal anaerobic bacteria.

The Role of the First 14 Amino Acids of Mature M1 Protein of Streptococcus pyogenes on Fibronectin-Binding Activity and Dimer Formation

KEMBAREN, ROGA FLORIDA, GANJARA, ADAM REZA, YURINA, VALENTINA, RETNONINGRUM, DEBBIE SOEFIE

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

Streptococcus pyogenes is one of the most important human pathogens which express a multi-facet of virulence factors on its cell surface.  One of the virulence factors that has been intensively-studied is the M protein that binds several human proteins.  M1 protein, a member of the M protein family, was previously found to bind human fibronectin (Fn), an activity that is responsible for bacterial internalization. A structural study showed that this protein consists of four regions: A, B, S, and C.  The study  was intended to investigate the role of the first 14 amino acid residues located at the non-helical region of M1 protein in binding Fn, and its ability to form a dimer. The DNA fragment encoding for the ABS protein lacking its first 14 amino acids (ABSD14aa) was cloned into pET-16b, overexpressed in Escherichia coli BL21(DE3), and the protein was purified by affinity chromatography. The purified protein was characterized by sodium dodecyl sulphate polyacrylamide gel electrophoresis and the Fn-binding activtiy was assayed by enzyme linked immunosorbent assay.  The result indicated that the M1 lacking its first 14 amino acids retains its dimerization and Fn-binding activities.

Establishment of Mycorrhizas of Four Different Species of Arbuscular Mycorrhizal Fungi Observed Using Petri-Dish Microcosm

MANSUR, IRDIKA, DODD, JOHN, JEFFRIES, PETER

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

The Petri-dish microcosm is a simple system to study the development of arbuscular mycorrhizal fungi (AMF) and has been shown to be suitable to observe the establishment of Acaulospora tuberculata BEG41, Gigaspora rosea BEG111, Glomus manihotis BEG112 and Scutellospora heterogama BEG40 on plant root. The non-agar based system could be easily used to observe the whole course of development of fungal-root interactions as well as the extra-radical structures of AMF without destructive sampling.  It was found that these AMF species were unique with regard to spore germination, root colonization, the architecture of the extra-radical mycelium (ERM), the spread of the ERM in the media, and sporulation.  The ERM of A. tuberculata BEG41and G. manihotis BEG112 spread mainly close to the root system, while the ERM of G. rosea BEG111 and S. heterogama  BEG40 could occupy areas unoccupied by roots.  The unique structure, the branch absorbing structure, was found in the four species of AMF, but differed in their shape and size

Genetic Diversity of Antifungi-Producing Rhizobacteria of Pseudomonas sp. Isolated from Rhizosphere of Soybean Plant

SUSILOWATI1, SUSILOWATI1, WAHYUDI, ARIS TRI, LESTARI, YULIN, WIYONO, SURYO, SUWANTO, ANTONIUS

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

Antifungi-producing rhizobacteria have been recognized playing an important role in plant disease suppression. In our laboratory, 13 indigenous soybeans´ rhizobacteria Pseudomonas sp. that showed strong growth inhibition of root pathogenic fungi, Rhizoctonia solani, Fusarium oxysporum and Sclerotium rolfsii, have been isolated from rhizosphere of soybean plant. For further understanding, the genetic diversity of the antifungi-producing Pseudomonas sp. was investigated using Amplified 16S rDNA Restriction Analysis (ARDRA) and 16S rRNA gene sequences analysis. 16S rDNA were amplified by PCR technique and digested with restriction endonuclease HaeIII, RsaI and AluI. Sequences of 16S rRNA gene were analyzed using the BLAST program for similarity searches on sequence databases. ARDRA based dendrogram analysis was carried out by neighbor-joining of TREECON 1.3b software package. ARDRA indicated the variability of Pseudomonas sp. based on the digestion sites. Dendrogram clustering analysis based on the restriction enzymes profile of the amplified rDNA distinguished Pseudomonas sp. into 7 ribotype groups. The sequences of 16S rRNA gene confirmed that the isolates belonging to Pseudomonas sp. and the phylogenetic tree formed 4 clusters. There was a quite overlap among ARDRA groups and 16S rRNA sequence clusters. This finding suggested that antifungal producing Pseudomonas sp. were present in the rhizosphere of soybean plant and the level of genetic diversity exist within these species. Sequence analysis of the 16S rRNA gene of the Pseudomonas sp. with an identical ARDRA pattern confirmed that members of an ARDRA group were closely related to each other.

Production of Exopolysaccharides by Strains of Streptococcus thermophilus

PURWANDARI, UMI, VASILJEVIC, TODOR

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

Two strains of Streptococcus thermophilus producing capsular and capsular-ropy exopolysaccharides (EPS) were examined for their growth and EPS production in M17 medium supplemented with glucose, galactose or lactose and incubated at 30, 37 or 42°C for 24 hours. Growth parameters (viable cells, OD, lactate production, pH) and EPS production were determined. Flow behavior of the EPS dispersions was assessed as a function of concentration and temperature. Culture growth during incubation was affected by types of sugar, temperature and time. Growth was enhanced by glucose, lactose and higher incubation temperature. EPS concentration in the medium was greater in the presence of glucose and galactose. Despite the restricted growth conditions, the capsular strain produced comparable levels of EPS to the capsular-ropy strain even under sub-optimal incubation temperature

Molecular Identification and Diversity of Yeasts Associated with Apis cerana Foraging on Flowers of Jatropha integerrima

BASUKRIADI, ADI, SJAMSURIDZAL, WELLYZAR, PUTRA, BANGGA BERISTAMA

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

There are only a few reports from tropical countries, and none from Indonesia, on yeasts associated with the Asiatic honeybee, Apis cerana. Here we report on yeasts associated with A. cerana foraging on flowers of Jatropha integerrima in the campus of the Universitas Indonesia, Depok, Indonesia. Yeasts were isolated from guts of 30 individual pollen-collecting bees (PCB) and nectar-collecting bees (NCB), and identified by their internal transcribed spacer (ITS) regions of their rDNA sequences. Based on ITS regions sequence data, 14 representative yeast isolates obtained from A. cerana were found to be closely related to Aureobasidium pullulans, Dothioraceae sp., Candida cf. apicola, C. cf. azyma, C. cellae, Metschnikowia sp., Kodamaea ohmeri and Yarrowia lipolytica. Undescribed yeast of the genus of Metschnikowia was also discovered in this study. At present, we assume there is association between C. cf. apicola and species closely related to C. cellae with A. cerana. Yeasts species associated with PCB differ from those found in NCB, indicating that PCB and NCB possess different and specific yeasts communities. Some yeasts species isolated from A. cerana show a low degree of similarity to their closest related species. Our study sheds light on the detection of several new taxa of yeasts associated with A. cerana.

Guide for Authors

Rusmana, Iman

Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

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