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Journal : Indonesian Journal of Biotechnology

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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Abstract

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
Effect of Oxidative Stress on AhpC Activity and Virulence in katG Ser315 Thr Mycobacterium tuberculosis Mutant Rintiswati, Ning; Wibawa, Tri; Asmara, Widya; Soebono, Hardyanto
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

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Abstract

AbstractMycobacterium tuberculosis strains resistance to INH is mainly caused by the alteration in several genesencoding the molecular targets. Mutation of katG at codon 315 especially Ser315Thr are responsible forINH resistance in a large proportion of TB cases. The aim of this study is to evaluate the influence of stressoxidative on AhpC activity of katG Ser315Thr of M.tuberculosis, and to find out the relation of AhpC and thevirulence of this mutant. The study design was laboratoric experimental, subjects of study were M.tuberculosisINH resistance strains, and the treatment were serial dose of H2O2. Eighty five M.tuberculosis INH resistantclinical strain were screened for mutation of katGSer315Thr by PCR/RFLP and characterized on the basis ofphenotypic properties (catalase activity and AhpC activity). AhpC activity of katG Ser315Thr M.tuberculosisstrains in response to oxidative stress condition was evaluated by culturing the strains on liquid culturemedium containing 1mM H2O2. To ascertain role of AhpC in the virulence of katGSer315Thr mutant strains, themutants were infected into human macrophages culture, and several indicator of virulence were observed (i.e:replication competence, and apoptosis induction on human macrophages). The results showed that katG Ser315Thr were identified in 23 (27,05%) of 85 INH resistance strains, all mutant strains had decrease of catalaseactivity. AhpC activity of katG Ser315Thr of M.tuberculosis increased significantly with increase of hydrogenperoxide dose. In addition , it has been shown that increased AhpC activity related to replication ability ofmutant, and reduction of apoptosis macrophages induction significantly. We conclude that the productionof AhpC of katG Ser315Thr M.tuberculosis induced by oxidative stress. There was a role of AhpC in virulenceof the M.tuberculosis katG Ser315Thr strains by replication capability and macrophages apoptosis.Keywords : katG Ser315Thr Mycobacterium tuberculosis- oxidative stress - AhpC - virulence
Cytotoxic Activity of Tegari (Dianella nemorosa Lam.) Methanol Extract Against HeLa Cells Karim, Aditya Krishar; ., Sismindari; Asmara, Widya; ., Istriyati
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Abstract

Dianella nemorosa Lam. also known as tegari belonging to the Liliaceae family. This plant has been utilized for Papua traditional medicine as well as anticancer agent. This research examined potential cytotoxic activity of tegari (D. nemorosa) leaves extract against cervical cancer cell line (HeLa). Methanol extract was obtained by extracting the leaves powder using methanol. Extract was then applied into HeLa cell line to find out the cytotoxic activity. MTT [3-(4,5-dimetilthiazol-2-il)2,5-difeniltetrazolium bromida) assay was used to measure the cytotoxic activity. The result indicated that D. nemorosa leaves extract possessed cytotoxic activity in HeLa cell line with IC50 values were 685,69 µg/ml, 506,43 µg/ml and 708 µg/ml at the incubation period of 24, 48 and 72 h respectively. The strongest cytotoxic was showed by methanol extract incubated in 48 h.
Isolation and Purifi cation of Chitinase Bacillus sp. D2 Isolated from Potato Rhizosfer Margino, Sebastian; Behar, Chatarina; Asmara, Widya
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Abstract

Potato Cyst Nematodes (Globodera rostochiensis) is one of the important potato’s pests and caused economic looses up to 70% in the several centrals of potato plantations in Indonesia. Potato Cyst Nematodes (PCN) shell component of egg shell containing chitin (inner layer) and vitelline/protein (outer layer), so the purpose of research was to fi nd out of chitin degrading bacteria for controlling of egg’s PCN by cutting of their life cycle. The results showed that Bacillus sp. D2 isolated from potato rhizosphere could produce extra cellular chitinase in the medium containing of 0.20% colloidal chitin and fermented for 72 hours. Result of chitinase purifi cation using ammonium sulphate precipitation and DEAE-Cellulose ion-exchange chromatography showed a specifi c activity 2691,052 U/mg and analyzing using SDS-PAGE 12.5% resulted in molecular weight 30 kDa. The apparent Km and Vmax of chitinase towards colloidal chitin were 2 mg/ml and 2.2 μg/h, respectively.  
Phylogenetic relationship of Gram Negative Bacteria of Enterobacteriaceae Family in the Positive Widal Blood Cultures based on 16S rRNA Gene Sequences Darmawati, Sri; Sembiring, Langkah; Asmara, Widya; Artama, Wayan T.; Kawaichi, Masashi
Indonesian Journal of Biotechnology Vol 19, No 1 (2014)
Publisher : Universitas Gadjah Mada

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Abstract

The purpose of this study was to analyze the phylogenetic relationship of Gram negative bacteria (3strains of Salmonella typhi, 1 strain of Escherichia coli, 1 strain of Serratia marcescens, and 3 strains of Enterobactercloacae) of Enterobacteriaceae family in positive Widal blood cultures based on 16S rRNA gene sequences. Theresults respectively showed that each two 16S rRNA gene clones of Serratia marcescens KD 08.4 had a closerelationship with 16S rRNA gene of Serrratia marcescens ATCC 13880 (similarity: 99.53-99.8%), Eschericia coliBA 30.1 with Eschericia coli ATCC 11775T (similarity: 99.38-99.67%), Salmonella typhi BA 07.4, Salmonella typhiKD 30.4, and Salmonella typhi SA 02.2 with Salmonella typhi ATCC 19430T (similarity: 99.4-100%) as well as theisolates of Enterobacter cloacae SA 02.1, Enterobacter cloacae BA 45.4.1, one 16S rRNA gene clone of Enterobactercloacae TG 03.5 with Enterobacter cloacae ATCC 23373 (similarity: 99.0-99.87%).
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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Abstract

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.
Antifungal Production of a Strain of Actinomycetes spp Isolated from the Rhizosphere of Cajuput Plant: Selection and Detection of Exhibiting Activity Against Tested Fungi A, Alimuddin; Widada, Jaka; Asmara, Widya; M, Mustofa
Indonesian Journal of Biotechnology Vol 16, No 1 (2011)
Publisher : Universitas Gadjah Mada

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Abstract

Actinomycetes are bacteria known to constitute a large part of the rhizosphere microbiota. Their isolation is an important step for screening of new bioactive compounds. Culturable actinomycetes populations from cajuput plant rhizosphere soils in Wanagama I Forest UGM Yogyakarta were collected to study about their antifungal activity. Among 17 of a total 43 isolates that showed activity were screened for producing antifungi substances. Screening for antifungal activity of isolates were performed with dual culture bioassay in vitro. One isolate that was designated as Streptomyces sp.GMR-22 was the strongest against all tested fungi and appeared promising for a sources of antifungal. Culture’s supernatant and mycelia were extracted with chloroform, ethyl acetate and methanol, respectively. Antifungal activity of crude extracts was tested by diffusion method against tested fungi. The result indicates that isolates of actinomycetes from cajuput plant rhizosphere could be an interesting sources of antifungal bioactive substances.
Purification and Characterization of Streptomyces sp. IK Chitinase Margino, Sebastian; Nugroho, Agustinus Joko; Asmara, Widya
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

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Abstract

Streptomyces sp. IK isolated from compost inoculants, could produce extra cellular chitinase in a medium containing 0.2% (w/v) colloidal chitin, fermented for 96 hours at 30oC. The enzyme was purified by a combination of ammonium sulphate precipitation and DEAE-Cellulose anion-exchange chromatography. On SDS-polyacrylamide gel electrophoresis analysis, the purified enzyme showed a mass of 71 kDa. Chitinase was optimally active at pH of 6.7 and at 37oC. Km value and Vmax of the protein for colloidal chitin were 2.92 mg/ml and 4.26 ìg/h, respectively.Key words : chitinase, Streptomyces, purification, characterization
Human Origin Lactobacillus casei Isolated from Indonesian Infants Demonstrating Potential Characteristics as Probiotics in vitro ., Widodo; Taufiq, Tiyas Tono; Aryati, Ety; Kurniawati, Asih; Asmara, Widya
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Abstract

The aim of this experiment was to isolate and identify Lactic Acid Bacteria (LAB) from infant faeces and subsequent evaluation of its potential probiotics. LAB was isolated from faeces of infants who consumed breast milk as the only source of diet on L-cysteine-supplemented MRS Agar, and incubated on 37oC for 48 hours. Colonies grew on this media were then identifi ed based on morphological, physiological and molecular approaches. Morphological and physiological identifi cations based on Gram staining, shape, motility, spore formation, catalase, CO2 and NH3 production, and the ability to grow on temperature at 10oC and 45oC. Molecular identifi cation based on the amplifi cation of 16S rRNA gene. The potential application of selected isolates for probiotics was evaluated based on the ability to grow on media with low pH and the addition of 0.5% bile salts, the ability to inhibit the growth of pathogenic Bacillus cereus and Eschericia coli, and in vitroadherence ability. On the basis of morphological, physiological and molecular analysis of 16S rRNA gene, it was concluded that the selected isolate 1AF was a strain of Lactobacillus casei. Evaluation of probiotic in vitro showed that 60.4% of cells were resistant to pH 2.0 for 90 minutes. Survival of isolate 1AF after growing at 0.5% bile salts was 70.8%. The selected isolate 1AF showed the ability to inhibit the growth of Eschericia coli and Bacillus cereus with inhibitory zone of 12.00±1,00 and 15.33±1.53 mm, respectively. In vitro study on the adherence value of isolate to solid plate was found at 46.5%. It is concluded that Lactobacillus casei isolate 1AF is a potential candidate as probiotics and subject to further in vivo evaluation.
Chemosystematic of Enterobacteriaceae Familia Obtained from Blood Cultures Based on Total Protein Profiles Darmawati, Sri; Sembiring, Langkah; Asmara, Widya; Artama, Wayan T.; Anwar, Syaiful
Indonesian Journal of Biotechnology Vol 18, No 1 (2013)
Publisher : Universitas Gadjah Mada

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Abstract

The purpose of this study was to determine the chemosystematic of 14 strains of bacteria in blood cultures from Semarang using 1 reference strain S. typhi NCTC 786, based on the total protein profi les with the similarity relationship analysis based on Simple Matching Coeffi cient (SSM) analysis and algorithm methodof unweighted pair group with averages (UPGMA) presented in a dendrogram. The results showed that thechemosystematic based on the total protein profi les using SDS-PAGE method can classify the member ofbacterial strains of each species. The Clusters respectively consist of 4 strains of S. typhi (similarity: 89.7%),2 strains of Ser. marcescens (similarity: 89.7%), two strains of E. coli, and one strain of Salmonella ssp, S. typhi NCTC 786 (similarity: 100%). Those three incorporated clusters had the similarity value of 75.3%. Those four strains of Ent. cloacae composed in one cluster (similarity: 100%) are incorporated in a cluster consisting of one strain of Kleb. pneumoniae (similarity: 92.9%). Both clusters were incorporated in a cluster consisting of S. typhi NCTC 786 (similarity: 67.9%).Key words: Enterobacteriaceae, chemosystematic, blood cultures, protein profile
Co-Authors . Sismindari ., Wdjijono A. Alimuddin Adi Heru Sutomo Aditya Krishar Karim Aditya Krishar Karim AETH. Wahyuni AETH. Wahyuni, AETH. Ag. Yuswanto Agnes Sri Harti Agnes Sri Harti Agnesia Endamg Tri Hastuti Wahyuni Agnesia Endang Tri Hastuti Wahyuni Agus Eko Srihanto Agus Eko Srihanto, Agus Eko Agustinus Joko Nugroho Agustinus Joko Nugroho, Agustinus Joko Aisida, Sufinatin Alma Linggar Jonarta Ambar Pertiwiningrum Andini, Syarifah Anwar Rosyidi April H Wardhana Arfian Rahma Shanti Artama , Wayan T. Arum Setiawan Arum Setiawan Asih Kurniawati Bambang Hariono Bambang Sumiarto Bambang Sutrisno Bancin, Girang Stevani Banun Kusumawardani Boy Bachtiar Br Sembiring, Samariana Budi Mulyaningsih Budi Mulyono BUDI SETIADI DARYONO Budi Setiadi Dayono Charles Rangga Tabbu Charles Rangga Tabbu Chatarina Behar Chatarina Behar, Chatarina Darmawati , Sri Darmayanti, Hervina Puspita Dewi Noor Hidayati Dewi Seswita Zilda Dito Anggoro Djaswadi Dasuki Doddi Yudhabuntara Dwi Aji Nugroho, Dwi Dyah Haryuningtyas Dyah Haryuningtyas Dyah Irnawati Eko Agus Srihanto Eko Agus Srihanto Eko Agus Srihanto, Eko Agus Elfayetti Elfayetti Elsa Kardiana Eni Harmayani Eni Harmayani Eni Yuniastuti Enny Yusuf Wachidah Yuniwarti Ety Aryati Ety Aryati, Ety Febriani Putri, Devita Fiska Salsabilla Gaol, Riski Fanni Lumban Gintung Patantis Gintung Patantis Gusti Ayu Yuniati Kencana Hafiza, Rohmania Zuhro Hamzah Muhammad Mardi Putra hanifah hanifah Hanny Hafiar Hardyanto Soebono Hardyanto Subono Harefa, Meilinda Suriani Harefa, Sri Aswinda Hari Eko Irianto Hasibuan, Novita Hasibuan, Novita Annisah Heru Susetya Hutauruk, Mutiara Cristeofani I Gusti Ketut Suarjana I Gusti Made Krisna Erawan I Gusti Ngurah Kade Mahardika I Wayan Suardana I. Istriyati Ida Tjahajati Iisnawati, Iisnawati Ika Dyah Kusumawati Ika Dyah Kusumawati, Ika Dyah Indwiani Astuti Istriyati ., Istriyati Istriyati Istriyati Istriyati Istriyati Istriyati, . Istriyati, . Iwan Dwiprahasto JAKA WIDADA Juni Handajani Kardians, Elsa Karna Wijaya Kerin Sisca Octaviani Luahambowo Khairunnisa Khairunnisa Khairunnisa Nasution Khrisdiana Putri Khrisdiana Putri, Khrisdiana Koichi Akiyama Krisdiana Putri Kumalasari, Eliana Dwi Langkah Sembiring LANGKAH SEMBIRING Langkah Sembiring Lina Sri, Widyawati Liza Angeliya Luahambowo, Kerin Sisca Octaviani M. Haryadi Wibowo M. Mustofa Mammed Sagi Mammed Sagi Mangkoewidjojo , Soesanto Marla Anggita Marpaung, Viviana Marsetyawan Soesatyo Marsetyawan Soesatyo Masashi Kawaichi Masashi Kawaichi, Masashi Maxs Urias Ebenheizer Sanam Meilinda Suriani Harefa Michael Haryadi Wibowo MM.Firdiana Krisnaningsih Mustofa M, Mustofa Mustofa Mustofa Nasution, Khairunnisa Niken Irfa Nastiti Ning Rintiswati Ning Rintiswati Nobuyuki Harada Nursyirwani, Nursyirwani Nurul Islamiyah Nuryono ., Nuryono Nyoman Reishita Andriyani Osman Sianipar Prasetyo, Dimas Bayu Purba, Tondang Raja Pangihutan Purnama Edy Santosa Purwoko, Agus Putra, Mulhady Raden Wisnu Nurcahyo Rahma, Mawaddah Rahmat Setya Adji Rahmat Setya Adji Regina TC Tandelilin Reni Nurjasmi Reni Nurjasmi, Reni Rini Widayanti Risbue Siregar Rosa Delima Lumbantungkup S Muharsini S Rahmah Umniyati, S Rahmah S. Sismindari Sabela, Serli Sagi , Mammed Salomo Hutahaean Salsabilla, Friska Santoso, Ferdinand Prayogo Cahyo Sarwo Edy Wibowo SATRIYAS ILYAS Sebastian Margino Sebastian Margino Sembiring , Langkah Sembiring, Samariana Sembiring, Samariana Br Setiawan , Arum Setiyono Setiyono Setiyono Setiyono Setyawan Budiharta Sidna Artanto Sidna Artanto Sidna Artanto, Sidna Simamora, Angelina Setia Simaremare, Ermas Simarmata, Claudia Grace Siregar, Risbue Sismindari . Siti Sunarintyas Sitti, Ummiyati Rahmah Situmorang, Duma Yanti Soemitro Djojowidagdo Soesanto Mangkoewidjojo Sri Darmawati Sri Darmawati Sri Darmawati Sri Lestari Subronto Prodjoharjono Sugeng, Mardihusodo Juwono Sugiharto Sulastri, Yati Surya Amanu Surya Amanu Surya Amanu Surya Amanu Susi Iravati Susi Iravati Susi Maulida, Susi Syaiful Anwar Syari’I Damanik, Muhammad Ridha Syukri Hidayat Titik Purwati Widowati Titik Purwati Widowati, Titik Purwati Tiyas Tono Taufiq Tondang Raja Pangihutan Purba Tri Ratnaningsih Tri Untari Tri Untari Tri Untari Tri Wibawa Triyanto . Tsutomu Nohno Vivina Marpaung W. Widjijono W. Widodo Wajar, Dony wayan T Artama wayan T Artama Wayan T. Artama Wayan T. Artama Wayan T. Artama Wayan Tunas Artama Wayan Tunas Artama Widagdo Sri Nugroho Widagdo Sri Nugroho Widodo Widodo Widya Hary Cahyati Wisnu Nurcahyo Wulandari, Anna Yahya, Adibah Yatri Drastini Yulita Kristanti Yundari, Yundari Yuni Wijayanti Yusro Nuri Fawzya Yusro Nuri Fawzya Zahwa, Afitzka Al Zilda, Dewi Zeswita