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Phylogenetic relationship of Gram Negative Bacteria of Enterobacteriaceae Family in the Positive Widal Blood Cultures based on 16S rRNA Gene Sequences Sri Darmawati; Langkah Sembiring; Widya Asmara; Wayan T. Artama; Masashi Kawaichi
Indonesian Journal of Biotechnology Vol 19, No 1 (2014)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (431.864 KB) | DOI: 10.22146/ijbiotech.8635

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%).
Adherence Pheno-genotypic of Escherichia coli O157:H7 Isolated from Beef, Feces of Cattle, Chicken and Human I Wayan Suardana; Wayan Tunas Artama; Widya Asmara; Budi Setiadi Dayono
Indonesian Journal of Biotechnology Vol 16, No 1 (2011)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1601.412 KB) | DOI: 10.22146/ijbiotech.15990

Abstract

Generally, adherence of micro-organisms to host cells is the frst step of the colonization to host surfaces.Escherichia coli O157:H7 can colonize to the intestine and induce attaching-effacing (AE) lessions. The capacityof inducing AE lesions is encoded by a pahtogenicity island, the locus of enterocyte effacement (LEE) thatcontains genes involved in generation of attaching and effaching (A/E) lesions. Among which that, the eaegene is encoding intimin, an outer membrane protein that is responsible to intimate attachment to the intestinalepithelial cells. A total of 20 local isolates obtained from human clinically, beef, cattle, chicken, and humannon-clinically were tested to adherence pheno-genotypic of E. coli O157:H7. The eae gene was identifed usingpolymerase chain reaction with a specifc primer i.e AE19 forward and AE20 reverse. To confrm phenotypicof gene, further study was performed by culturing the bacteria in vero cell, followed by Giemsa staining andAcridine Orange Fluorescent staining 3 h and 6 h after incubation, respectivelly. Result of study showed thatthere were 19 out of 20 (95%) isolates identifed positive eae gene. Giemsa staining appeared that the bacteriawith positive eae gene performed a cluster arroud of cell (localized adherence). On the other hand, the negativeeae gene appeared as a diffuse adherence (DA). The study indicated that almost all of E. coli O157:H7 localisolates which was positive eae gene had potency to colonize to the intestine and induce attching-effachinglessions, and also cause cytopahatic effects in intestinal epithelial cell
Effect of Oxidative Stress on AhpC Activity and Virulence in katG Ser315 Thr Mycobacterium tuberculosis Mutant Ning Rintiswati; Tri Wibawa; Widya Asmara; Hardyanto Soebono
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (287.371 KB) | DOI: 10.22146/ijbiotech.16369

Abstract

Mycobacterium 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.
Detection and identifcation of adherence genes of intestinal-origin Lactobacillus and Pediococcus strains grown on gastric mucin in vitro W. Widodo; Sri Lestari; Widya Asmara
Indonesian Journal of Biotechnology Vol 20, No 2 (2015)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (858.388 KB) | DOI: 10.22146/ijbiotech.24346

Abstract

One of the primary selection criteria for potential probiotics is the ability to adhere to the host gastrointestinal tract. This study evaluated the in vitro adhesion ability on gastric mucin of two Lactobacillus casei strains (AP and AG) and two Pediococcus acidilactici strains (BE and BK), and identifed the corresponding genes responsible for adherence. Adhesion assays were performed in 96-well polystyrene microtiter plates using gastric mucin from porcine stomach as the matrix. An in vitro study on gastric mucin revealed that lactobacilli had a greater adherence ability compared with pediococci strains. The potential adherence genes were investigated using polymerase chain reaction (PCR) technology. Using specifc primers, PCR studies amplifed 150 base pairs of a potential mub gene and 161 base pairs of a potential ef-Tu gene, but no amplifed bands for potential map and bac genes were obtained. Sequence comparisons showed that the 150 and 161 amplifed base pairs were respectively homologous to the mub of Pediococcus acidilactici and ef-Tu genes of Lactobacillus paracasei. We concluded that the adherence ability of two strains of Lactobacillus casei (AP and AG) and two strains of Pediococcus acidilactici (BE and BK) on gastric mucin is in accordance with the presence of ef-Tu and mub genes. High level attachment in lactobacilli is likely to correlate with the ef-Tu gene, which is a lactobacilli-specifc adhesive gene.
Evaluation of rapid detection kit against avian influenza A virus and H5 subtype for field Sample Michael Haryadi Wibowo; Tri Untari; Sidna Artanto; Krisdiana Putri; Surya Amanu; Widya Asmara
Indonesian Journal of Biotechnology Vol 21, No 1 (2016)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (846.082 KB) | DOI: 10.22146/ijbiotech.26792

Abstract

Avian influenza virus is poultry viral disease, which causes high economic losses. Various efforts have been made to control the disease. One effort is required screening fast and precise diagnostic test. This study was aimed to determine the potential of rapid test kit of AIV/H5 Anigen Rapid Test for the detection of AI virus types A and subtype H5 in the field. Some tests were carried out, e.g.: the potential test, cross-reaction test, sensitivity and specificity test. Potency test was done to evaluate potential of detection limits of the kit, by having the test of serial dilution of AI virus positive control. Cross-reaction test was done to detect antigens other than AI virus H5N1, e.g.:  IB virus of Massachuset strain, IBV strain 4-91, Newcastle Disease virus, and Escherichia coli. Sensitivity and specificity test were applied to the filed samples which clinically and laboratory were confirmed as H5N1 positive. To confirm the result of rapid test was then being done by reverse transcriptase polymerase chain reaction. Based on these results it can be concluded that, Anigen Kit AIV/H5 Ag Rapid Test can detect antigen-containing samples having AI virus HA titer up to 26of type A virus, and up to 25 for subtype H5 virus. Anigen Kit AIV/H5 Ag Rapid Test showed no cross-reactions with Infectious Bronchitis virus, Newcastle Disease virus, and Escherichia coli. Anigen A Rapid Test Kit AIV Ag showed a sensitivity of 50% and specificity of 100%, while Anigen Ag Rapid Test Kit AIV/H5 showed a sensitivity of 25% and specificity is 100%.
The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia Dewi Noor Hidayati; Eko Agus Srihanto; Tri Untari; Michael Haryadi Wibowo; Koichi Akiyama; Widya Asmara
Indonesian Journal of Biotechnology Vol 24, No 1 (2019)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (14.17 KB) | DOI: 10.22146/ijbiotech.44298

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Considering the increasing incidence of infectious bovine rhinotracheitis (IBR) in Indonesia, it was necessary to conduct a more in-depth study of bovine herpesvirus-1 (BHV-1) as the causative agent of IBR disease. Previous research reports indicate that the BHV-1 subtypes found in Indonesia are subtype 1.1. Currently, IBR field case detection in Indonesia still uses the serological method (ELISA), which has the potential to give false positive results and cannot explain the virus subtype. Other detection methods, such as viral isolation, take longer and require adequate resources. This study aimed to determine the BHV-1 subtypes of Indonesian isolates using molecular techniques. Nested PCR using two pairs of primers was successfully used to amplify the glycoprotein D (gD) gene. The gD gene fragment was cloned into the pGEM-T plasmid. Analysis of the gD gene sequence was subsequently carried out to determine the BHV-1 character of the Indonesian isolates. The results indicated that the isolates were different from the previous isolates, and had similarities (100%) with subtype 1.2 strain SP1777 and SM023.
PROFIL PLASMID Escherichia coli RESISTEN TERHADAP BEBERAPA ANTIBIOTIKA YANG DIISOLASI DARI PETERNAKAN AYAM KOMERSIAL Michael Haryadi Wibowo; Widagdo Sri Nugroho; Widya Asmara
Jurnal Sain Veteriner Vol 29, No 1 (2011): JUNI
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (5451.848 KB) | DOI: 10.22146/jsv.290

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Penelitian ini bertujuan untuk mengetahui profil plasmid E.coli yang resisten terhadap ampisilin, streptomisin dan enrofloksasin. Delapan isolat E.coli yang telah diidentifikasi dan diuji sensitivitasnya terhadapketiga antibiotik tersebut, selanjutnya diisolasi plasmidnyas. Isolat E. coli dipupuk pada kaldu laktosa diinkubasi dalam shaker incubator pada suhu 370 C semalam. Sel dipanen dengan sentrifugasi 12.000 rpm, selama 5 menit sebanyak dua kali. Isolasi plasmid dilakukan dengan metode lisis alkali menggunakan larutan lisis I, II, dan III. Presipitasi plasmid dilakukan dengan 3 M Na asetat dan ethanol absolut. Profil plasmid dibaca pada agarosa 1%, setelah dilakukan elektroforesis menggunakan marker plasmid. Hasil penelitian menunjukkan profil plasmidDNA E. coli tersebut teramati sebagai pita-pita DNA yang berpendar oleh pendedahan sinar ultraviolet. Plasmid yang terisolasi mempunyai ukuran yang sangat besar atau mega plasmid yaitu terletak pada 4268 bp, 4873 bp, 5148 bp dan terletak diantara 5148 bp dan 21.226 bp. Masing-masing isolat E. coli memiliki jumlah plasmid yang bervariasi antar 1 sampai 3 plasmid DNA.Kata kunci: Escherichia coli, resistensi, profil plasmid
Isolasi dan Propagasi Agen Penyebab Penyakit dari Kasus Terdiagnosa Penyakit Infektious Laryngotracheitis (ILT) pada Telor Ayam Berembrio Michael Haryadi Wibowo; Widya Asmara
Jurnal Sain Veteriner Vol 20, No 2 (2002): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1739.96 KB) | DOI: 10.22146/jsv.303

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ISOLATION OF VT1 AND/OR VT2 GENE-BEARING Escherichia coif FROM CATTLE, SWINE AND SHEEP AND GOAT = ISOLASI Escherichia coli PEMBAWA GEN VT1 DAN VT2 DART SAPI, BABI DAN DOMBA/KAMBING Yatri Drastini; Setyawan Budiharta; Widya Asmara
Jurnal Sain Veteriner Vol 20, No 2 (2002): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1493.612 KB) | DOI: 10.22146/jsv.314

Abstract

Dewasa ini dikenal adanya lima kelas EScherichia coli enterovirulen, termasuk E. coli verositotoksigenik (VTEC). Nama VTEC berhubungan dengan verositotoksin yang dihasilkan, yang disandi oleh gen VT dalam kromosom E. coli. Identifikasi VTEC pada awalnya selalu dikaitkan dengan serotipe 0157:H7. Tulisan ini melaporkan isolasi E. coli pembawa gen VT1 dan/atau VT2 dari sapi, babi, dan domba/kambing. Oles rekturn diambil dari hewan, dan adanya E. coli dideteksi dengan BGA, EMBA dan SMAC. Uji aglutinasi lateks digunakan untuk mendeteksi antigen 0157, dan adanya gen VT1 dan VT2 dideteksi dengan PCR. Paling tidak 44% isolat E. coli dari sapi tidak inernfermentasi sorbitol dengan 1,5% di antaranya adalah serotipe 0157. Isolat E. coil pembawa gene VTI danlatau VT2 paling banyak berasal dari babi. Namun, tidak semua E. coil pembawa gen tersebut adalah 0157.
Isolasi dan Identifiicasi Serologis Virus Avian Influenza Dari Sampel Unggas Yang Diperoleh di D.I. Yogyakarta dan Jawa Tengah = Isolation and Serological Identification of Avian Influenza Virus From Poultry Sample ... Michael Haryadi Wibowo; Widya Asmara; Charles Rangga Tabbu
Jurnal Sain Veteriner Vol 24, No 1 (2006): JUNI
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3152.733 KB) | DOI: 10.22146/jsv.346

Abstract

Avian Influenza (AI) merupakan penyakit penting pada unggas, karena dapat menyebabkan kerugian ekonomi secara signifikan, dengan tingkat morbiditas dan mortalitas penyakit sangat tinggi. L,ebih dan itu potensi penularan penyakit AI dari hewan ke manusia, memberikan dampak ekonomi tersendiri. Beberapa kasus yang diduga sebagai AI banyak mewabah di beberapa daerah di Indonesia. Penyakit tersebut cukup membingungkan peternak dan sangat dikacaukan dengan penyakit Newcastle (ND) karena kedua penyakit mempunyai kemiripan karakter dan gejala klinis. Penelitian ini bertujuan untuk mengkonfirmasi apakah wabah penyakit tersebut disebabkan oleh virus AI atau virus ND. Sampel isolasi diambil dari paru atau trakhea, kemudian diproses lebih lanjut untuk diisolasi, dipropagasi secara in ovo menggunakan telur ayam berembrio umur 9 sampai 12 hari, spesific pathogen free atau telur yang setidaknya bebas antibodi terhadap virus AI. Teknik isolasi menurut standar prosedur Office International des Epizooties (01E) dan kemungkinan adanya pertumbuhan virus diuji terhadap kemampuan mengaglutinasi sel darah merah ayam atau hemaglutinasi (HA). Uji HA positif, mengindikasikan ada pertumbuhan virus ND atau virus AL Kedua jenis virus tersebut dapat dibedakan dengan uji hemaglutinasi inhibisi (HI) menggunakan serum anti dari masing-masing virus yang diuji. Berdasarkan hasil penelitian ini dapat diambil suatu kesimpulan bahwa beberapa sampel unggas, yaitu: ayam petelur, ayam broiler, ayam kampung, dan burung puyuh, yang di peroleh dari beberapa daerah di D.I. Yogyakarta dan Jawa Tengah dan secara klinis menunjukkan gejala tersifat maupun tidak tersifat AI, secara serologis dapat dikonfirmasi sebagai virus avian influenza sub-tipe 145Ni.
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 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 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 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 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 Br Setiawan , Arum Setiyono Setiyono Setiyono Setiyono Setyawan Budiharta Sidna Artanto Sidna Artanto Sidna Artanto, Sidna Simaremare, Ermas Simarmata, Claudia Grace Siregar, Risbue Sismindari . Siti Sunarintyas Sitti, Ummiyati Rahmah 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