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Articles by issue : Vol 8, No 2 (2014): June 2014
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Articles
Diversity of Lactic Acid Bacteria Isolated from Indonesian Traditional Fermented Foods

MUSTOPA, APON ZAENAL, FATIMAH, FATIMAH

Microbiology Indonesia Vol 8, No 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

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Abstract

The diversity of lactic acid bacteria was evaluated from Indonesian fermented foods such as dadih (buffalo fermented milk), tempoyak (fermented durian), bekasam (fermented meat), and tape ketan (fermented glutinous rice). Lactic acid bacteria were enumerated using selective media and characterised based on a genotypic methods such as rep- PCR and RAPD-PCR, as well as 16S rRNA gene sequencing of representative strains. Forty-six colonies had successfullybeen isolated from Indonesian fermented foods. The great majority of these colonies originated from dadih (43.48%), tempoyak (39.13%), bekasam (13.04%) and tape (4,3%). The 46 isolates were characterised based on a genotypic methods such as RAPD and rep-PCR as well as 16S rRNA gene sequencing of representative strains. The rep-PCR result yielded seven clusters (I-VII) at a similarity level of 75-88% and RAPD-PCR used LB2 primer, M13 primer and primer A, B, C. The RAPD result using LB2 primer yielded eight clusters (I-VIII) at a similarity level of 82-91%. Identification using 16S rRNA showed that the majority strains as Lactobacillus plantarum, Lactobacillus fermentum and Pediococcus pentosaceus strains.

Determination of blaVIM and blaIMP Resistant Genes to Meropenem on Pseudomonas Aeruginosa Isolates in Hospitalization Bronkopneumonia Patient at Internal Medicine HCU of RSUP DR. M. Djamil Padang

NOVELNI, RINGGA, MARLINA, MARLINA, RAVEINAL, RAVEINAL

Microbiology Indonesia Vol 8, No 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

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Abstract

In this research, sample were taken prospectively to the hospitalization patients at the Internal Medicine HCU of RSUP DR. M. Djamil Padang. This research was begun with bacteria isolating of P. aeruginosa of the sputum samples of patients who suffered bronkopneumonia. The isolation were started with samples cultivation to the Cetrimide Agar media which was a selective media for bacteria of P. aeruginosa. To determined the species of the bacteria, it were identified with Gram staining, TSIA test, citric test, urease test, MR/VP test and detection of 16S rRNA genes. The isolation and identification result showed that from 20 sputum samples of patients there were just 10 (50%) samples were positive for P. aeruginosa bacteria. The positive isolates of P. aeruginosa were continued   to detection of blaVIM and blaIMP.resistant genes to meropenem respectively. The result showed that Metallo-β-Lactamase (MBLs) enzyme positively for all of P. aeruginosa were resistant to meropenem.

Cloning and Gene Expression of AnsZ Encoding L-Asparaginase Enzyme from Local Bacillus sp.

AZARA, RIMA, HELIANTI, IS, KUSNADI, JONI, YUNIANTA, YUNIANTA

Microbiology Indonesia Vol 8, No 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

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Abstract

L-asparaginase is an enzyme that catalyzes the hydrolysis of L-asparagine into L-aspartic acid and ammonia. In medical aspect, L-asparaginase especially those came from E. coli and Erwinia chrysanthemi used as chemotherapy agent of acute lymphoblastic leukemia (ALL). However, new potential organisms possessing L-asparaginase production capacity with a similar therapeutic effect are still required . In Bacillus subtilis strain 168, there are two kinds of L-asparaginase gene, AnsA and AnsZ. The study of the later L-asparaginase (AnsZ) has not been conducted intensively. The aim of this study is, first, to isolate this gene of L-asparaginase (AnsZ) from local Bacillus sp. and then to express this gene in Escherichia coli. Using PCR-cloning method, an open reading frame (ORF) containing 1128 bp was obtained. The ORF has 99% homology with sequence of L-asparaginase from Bacillus subtilis Bsn5. The gene then was subcloned into pET 21d (+) with his6-tag in the C-terminal of the gene product and expressed in E.coli BL21. L-asparaginase activity analyses showed that recombinant E. coli containing recombinant plasmid with open reading frame (ORF) L-asparaginase (AnsZ) from Bacillus subtilis had higher activity than that is not containing ORF L-asparaginase (AnsZ). Purification with HisPur TM Ni-NTA Purification Kit increased the specific activity of L-asparaginase (AnsZ) enzyme to 29 fold.

Effect of Micro-encapsulated Synbiotic at Different Frequencies for Luminous Vibriosis Control in White Shrimp (Litopenaeus vannamei)

MUNAENI, WAODE, YUHANA, MUNTI, WIDANARNI, WIDANARNI

Microbiology Indonesia Vol 8, No 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

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Abstract

The aim of this study was to evaluate the effect of micro-encapsulated synbiotic application at different frequencies for luminous disease control in white shrimp (Litopenaeus vannamei). The luminous disease is caused by Vibrio harveyi. In this experiment, a synbiotic which was a combination of the probiotic Bacillus sp. NP5 RfR and the oligosaccharides from sweet potato (Ipomea batatas L.) jago variety was apllied. The synbiotic was encapsulated by spray drying method. The in vivo experiment was conducted by supplementing the shrimp’s diet with the micro-encapsulated synbiotic for 40 days. Treatments included the administration micro-encapsulated synbiotic in different frequencies i.e. once a week (A), twice a week (B), daily (C), and without micro-encapsulated synbiotic (control treatment). The control treatment consisted of positive (K+) and negative (K-) controls. After 30 days period, all of the shrimp were challenged by intramuscular injection of pathogenic V. harveyi RfR at a concentration of 106 CFU ml-1 except the negative control. The treatment C resulted in significantly higher survival rate (SR), specific growth rate (SGR), and immune responses than those of the controls, whereas the feed conversion ratio (FCR) was lower than the controls. In addition, the population of Bacillus sp. NP5 RfR and total bacterial count (TBC) in the intestines increased, whereas the population of V. harveyi RfR and the total vibrio count (TVC) were lower compared to the controls.

Exploration, Isolation and Quantification of β-carotene from Bacterial Symbion of Acropora sp.

WUSQY, NAELY K., LIMANTARA, LEENAWATY, KARWUR, FERRY F

Microbiology Indonesia Vol 8, No 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

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Abstract

In the microbial world, pigments are one of the most conspicuous traits. Marine bacteria associated with Acropora sp. collected from Taka Cemara, Karimunjawa Islands were screened for the production of a yellow pigment. The isolation of bacterial symbionts from Acropora sp. on Zobell 2216E medium resulted in one bacterium, KJ5, positively synthesized carotenoids. By reverse phase HPLC analysis, one peak of the pigment types was identified as a β-carotene peak which appeared at 60.24 min. Then, sample of the β-carotene was collected and identified according to their spectral characteristics and compared with the published data in different types of solvent. Based on the HPLC analysis, the total β-carotene contents were calculated by converting the broad absorption of β-carotene. Molecular identification of the bacterium KJ5 using 16S rDNA showed that bacterium KJ5 was closely related to Erythrobacter flavus with 96% homology value.

Population and Diversity of Endophytic Bacteria Associated with Medicinal Plant Curcuma zedoaria

SULISTIYANI, TRI RATNA, LISDIYANTI, PUSPITA, LESTARI, YULIN

Microbiology Indonesia Vol 8, No 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

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Abstract

Traditionally Curcuma zedoaria (white turmeric) known as herbal medicine which possessing many biological activities. Many endophytic bacteria live in association with their host and may play an important biological roles. The main interest of this study was to investigate the endophytic bacterial diversity associated with white turmeric. White turmerics were collected from three locations in Bogor, West Java, Indonesia. The isolation of endophytic bacteria was carried out using 4 kinds media (Nutrient Agar (NA), NA contained white turmeric extract (NAT), Water Yeast Extract Agar (WYEA), WYEA contained white turmeric extract (WYEAT)), and 2 methods of spread plate and plant piece methods. The identification of selected isolates was conducted by molecular analysis based on 16S DNA. The suitable media and method of isolation endophytic bacteria were NA and spread plate method. A total of 207 bacterial colonies were isolated from rhizomes, stems, and leaves and 73 endophytic bacteria were selected based on morphological characteristics. From them, 32% isolates from Bojong Gede, 22% isolates from Cibinong and 46% isolates from Dramaga were obtained. Endophytic bacteria were predominated 38% in the rhizomes, 32% of stems, and 30% of leaves. Based on 16S rDNA sequence analysis, the isolates were belonging to the cluster Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria, with twenty three different genera includes Stenothropomonas, Pseudomonas, Enterobacter, Providencia, Klebsiella, Dickeya, Pantoea, Bacillus, Acinetobacter, Citrobacter, Mycobacterium, Cellulomonas, Microbacterium, Methylobacterium, Penylobacterium, Roseomonas, Agrobacterium, Bosea, Xanthobacter, Rhizobium, Burkholderia, Ralstonia, and Alcaligenes. The plant location, age, part of plant, media and method of isolation seem to influence the endophytic bacterial communities.

Role of Chloramphenicol Acetyltransferase (CAT) Enzyme for Early Detection of Chloramphenicol Resistant Salmonella typhi

NURTJAHYANI, SUPIANA DIAN

Microbiology Indonesia Vol 8, No 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

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Abstract

Microbial resistance to antibiotic is a major problem in the treatment of infectious diseases. The purpose ofthis study was to identification the role of CAT enzyme for early detection of chloramphenicol resistant. Observation was performed on isolated from Dr. Sutomo Hospital, Surabaya Health Laboratory, and Microbiology Laboratory-Airlangga University, Surabaya in 2009 and had been subcultured.The sub-culture was then tested for its susceptibility to chloramphenicol by using anti-chloramphenicol acetyl transferase (CAT) antibody from Sigma (catalog number C.9336). Susceptibility test using dilution and diffusion methods proved that resistant could change to sensitive and vice versa. It seems that the CAT enzyme can bind anti-CAT so reversing the resistant into sensitive and conversely, sensitive into resistant one.