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PROFIL PLASMID Escherichia coli RESISTEN TERHADAP BEBERAPA ANTIBIOTIKA YANG DIISOLASI DARI PETERNAKAN AYAM KOMERSIAL Haryadi Wibowo, Michael; Sri Nugroho, Widagdo; Asmara, Widya
Jurnal Sain Veteriner Vol 1, No 29 (2011)
Publisher : Fakultas Kedokteran Hewan

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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
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 Drastini, Yatri; Budiharta, Setyawan; Asmara, Widya
Jurnal Sain Veteriner Vol 20, No 2 (2002)
Publisher : Fakultas Kedokteran Hewan

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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 Propagasi Agen Penyebab Penyakit dari Kasus Terdiagnosa Penyakit Infektious Laryngotracheitis (ILT) pada Telor Ayam Berembrio Wibowo, Michael Haryadi; Asmara, Widya
Jurnal Sain Veteriner Vol 20, No 2 (2002)
Publisher : Fakultas Kedokteran Hewan

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PATOGENISITAS ISOLAT Escherichia con POSITIF CONGO RED PADA TELUR AYAM BEREMBRIO UMUR 12 HARI = PATOGENICITY OF CONGO RED POSITIVE ISOLATE OF Escherichia coil IN THE 12-DAYS OLD CHICKEN EMBRYOS Nugroho, Widagdo Sri; Wibowo, M. Haryadi; Asmara, Widya
Jurnal Sain Veteriner Vol 20, No 1 (2002)
Publisher : Fakultas Kedokteran Hewan

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Penelitian ini bertujuan untuk mengetahui patogenisitas isolat Escherichia coif positif congo red. Escherichia coil dari kasus kolibasilosis ayam diisolasi menggunakan media TSA dan EMB kemudian kemampuan pengikatan warna congo red diuji dengan agar congo red (TSA+0,003% congo red). Tingkat patogenisitas isolat yang mengikat warns congo red dilihat dari uji kematian embrio. Empat isolat E.coli positif conga red (CR+) dan 1 isolat negatif terhadap congo red (CR -) diinokulasikan pada telur berembrio umur 12 hari. Tingkat kematian embrio selama arum bad pascainokulasi masing-masing isolat E.coli positif congo red (500 colony form unit / CM) pada kantung alantois telur berembrio umur 12 hari berbeda antar kelompok. Angka kematian yang diperoleh dari isolat CR + 1, 2, 3, dan 4 masing-masing adalah 10%, 20%, 60% dan 100%. Perubahan anatomi yang tampak yaitu terjadinya perdarahan kulit pada embrio yang mati dan secara mikroskopik lesi-lesi pada hati, jantung, dan limpa menujukkan adanya septisemi. Isolasi dan uji ulang congo red terhadap inokulat positif conga red memperlihatkan bahwa beberapa isolat kehilangan kemampuan mengikat warna conga red. Isolat-isolat tersebut memiliki angka kematian yang rendah (10-20%). Variasi kemampuan isolat mengikat warm conga red memiliki keterkaitan dengan patogenisitasnya.
KARAKTERISASI MOLEKULER DAN UJI PATOGENESITAS STREPTOCOCCUS PATOGEN ISOLAT ASAL BALI Suarjana, I Gusti Ketut; Asmara, Widya
Buletin Veteriner Udayana Vol. 4 No.1 Pebruari 2012
Publisher : Buletin Veteriner Udayana

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The main of this study were characterized of muramidase released protein (MRP) andexctracellular factor (EF) as virulence factor of Streptococcus beta hemolityc of Bali islotesand its patogenecity on mice.The MRP was isolated from the cell walls bacteria withmuramidase (lysozyme) and EF was obtained from supernatant of bacteria precipated with70% ammonium sulphate and then dialysed. These protein were identified by usingsodium-dodecyl sulphate polyacrylamide gel electrophoresis ( SDS-PAGE). Isolates thatwas observed are five consist of three isolates from pigs and two isolates from monkeys.Pathogenecity test using 20 mices divided into four group. Group I inoculated with 0,1 mltodd-hewitt broth steril as negative control, group II inoculated with 0,1 ml inoculum ofbacteria of Streptococcus suis type 2 (strain D282) as positive control, group III inoculatedwith 0,1 ml Streptococcus beta haemolytic isolated from monkey and group IV inoculated with 0,1 ml Streptococcus beta haemolytic isolated from pig. The result of this studyshowed that all isolates were consist of eight protein bands of MRP and one EF of 110 kDamolecular weight. Eight protein MRP were 125 kDa, 76 kDa, 60 kDa, 57 kDa, 48 kDa, 45kDa, 30 kDa, and 28 kDa respectively. Each isolates had two major protein bands of MRP(76 kDa and 45 kDa).The patogenecity test in mice showed that the morbidity andmortality rates were 100% and 60% respectively. The prevalence of meningitis in mice are100%. Clinical sign were observed 30 hour post inoculated (pi) whereas mice found deathstarty 48 pi.
AMINO-TERMINUS OF POLYMERASE BASIC-2 OF AVIAN INFLUENZA VIRUS OF H5N1 SUBTYPE ISOLATED FROM VARIOUS ANIMAL SPECIES IN INDONESIA Yuniati Kencana, Gusti Ayu; Asmara, Widya; Rangga Tabbu, Charles; Kade Mahardika, I Gusti Ngurah
Jurnal Veteriner Vol 9, No 3 (2008)
Publisher : Jurnal Veteriner

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The information on pathogenicity and adaptation factors of avian influenza virus (AIV) in mammalsis very inportant in an effort to reduce the risk of avian influenza (AI) pandemic in the future. Polymerasegene complex appears to be the major factors for adaptation of AIV to certain animal species. A preliminarystudy on role of non-coding region (NCR) and amino-terminus of polymerase-basic 2 (PB2) is presented.Purified viral RNA of AIV isolated from chicken, duck, pig, and quail of Bali and Yogyakarta was reversetranscribed into cDNA and amplified using reverse transcriptase-polymerase chain reaction (RT-PCR)using PB-2 universal forward primer and specifically designed backward primer. The result showed thatall AIV’s H5N1 isolated from chicken, duck, quail, and pig, posed PB2 amino-terminus typical for IndonesianAIV H5N1. However, polymorphic amino acids of the protein fragment did not show any species specificmotive, with the exception of the pig isolate Sw/Tabanan/2006 which had specific substitution of D16E,H17Q, M40I, and H124Y.
VARIATION OF NON-CODING REGION AND CODING REGION OF 5’-TERMINAL CRNA OF POLYMERASE BASIC 1 OF AVIAN INFLUENZA VIRUS SUBTYPE H5N1 Yuniati Kencana, Gusti Ayu; Asmara, Widya; Rangga Tabbu, Charles; Kade Mahardika, I Gusti Ngurah
Jurnal Veteriner Vol 10, No 1 (2009)
Publisher : Jurnal Veteriner

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The sequence of the Non-Coding Region (NCR) and Coding Region (CR) of 5’-terminal cRNA of thepolymerase basic 1 (PB1) gene as a major factor for the species adaptation of avian influenza virussubtype H5N1 (AIV H5N1) has been analysed. The information could be a virological signal for theemergence of a new strain with pandemic potential. Total RNA from twenty six (26) avian influenzasubtype H5N1 isolates were amplified using reverse-transcriptase-polymerase chain reaction (RT-PCR)with a universal forward primer for influenza virus and specifically designed backward primers. Fifteen(15) PB1 gene fragments could be amplified. RT-PCR products were sequenced and analyzed using Mega4software. The length of NCR of PB1 gene was found to be 24 bases and mostly shows conserved sequence,with an exception of Dk/Badung/2006 isolate which has C-7T substitution. A/T composition of PB1 NCRwas 54,2%, while the Dk/Badung/2006 isolate was 58,3%. Species and geographical specificity could not befound in the genetic distance, the amino acid polymorphism, as well as the phylogenetic analysis of t
Non Coding Region dan Amino Terminus Gen Polimerase Asidik Virus Avian Influenza Subtipe H5N1 Asal Hewan Indonesia Yuniati Kencana, Gusti Ayu; Asmara, Widya; Rangga Tabbu, Charles; Kade Mahardika, I Gusti Ngurah
Jurnal Veteriner Vol 11, No 3 (2010)
Publisher : Jurnal Veteriner

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The knowledge on the species adaptation factor of avian influenza virus of H5N1 subtype (AIV H5N1)is very important as a signal for the emergence of a new strain with pandemic potential. This research wasconducted to find out the sequence variation of the Non-Coding Region (NCR) and Coding Region (CR) of 5’-terminal cRNA of the polymerase acidic (PA). Total RNA from twenty six (26) avian influenza virussubtype H5N1 isolates were amplified using reverse-transcriptase-polymerase chain reaction (RT-PCR)with a universal forward primer for influenza virus and specifically designed backward primers. Nineteen(19) gene fragments of PA could be amplified. RT-PCR products were sequenced and analyzed using Mega4 software. The length of NCR of PA gene was found to be 24 bases and conserved. A/T composition of PANCR was 58.3%. Species and geographical specificity could not be found in the genetic distance, the aminoacid polymorphism, as well as the phylogenetic analysis of the CR. RNA sequencing is discussed andrecomended to be studied further.
IDENTIFIKASI ESCHERICHIA COLI O157:H7 SERTA DETEKSI GEN SHIGA LIKE TOXIN 1 DAN 2 ASAL FESES HEWAN, DAGING, DAN FESES MANUSIA Suardana, I Wayan; Tunas Artama, Wayan; Asmara, Widya; Setiadi Daryono, Budi
Jurnal Veteriner Vol 11, No 4 (2010)
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Escherichia coli O157:H7 with the ability to produce shiga-like toxin was isolated from beef, cattle, chicken, and human feces. Due to its importance to human health, it is necessary to identify the genes encoding the production of shiga-like toxin, stx1 and stx2 respectively to further understand the pathogenesis. Isolation of E. coli was done on Eosin Methylene Blue Agar (EMBA), followed by identification on Sorbitol MacConkey Agar (SMAC), latex agglutination test, and H7 antiserum test, respectivelly. The existence of genes stx1 and stx2 in E. coli O157:H7 was confirmed molecularly using PCR method with specific primers LP 30/31 and LP 43/44, Stx2 (F)/Stx2 (R) respectively. Escherichia coli O157:H7 was isolated from 22 out of 344 samples (6,4%). Some isolates showed gene stx1 and stx2 was detected in two isolates as indicated by a 384 bp band (stx1 gene), 584 bp and 1588 bp bands (stx2 gene) respectivelly. The results indicated that local isolates E. coli O157:H7 are potential as a zoonoses agent.
Studi Epidemiologi Agen Zoonosis Escherichia coli O157:H7 melalui Analisis Random Amplification of Polymorphic DNA (RAPD) Suardana, I Wayan; Tunas Artama, Wayan; Asmara, Widya; Setiadi Daryono, Budi
Jurnal Veteriner Vol 12, No 2 (2011)
Publisher : Jurnal Veteriner

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Epidemiological studies of zoonotic agent Escherichia coli O157:H7 have been analyzed pheneticallyand or phylogenetically. In a phenetic classification, micoorganisms are arranged into groups (phena) onthe basis of high overall similarity using both phenotypic and genotypic methods without judgementaspect of its ancestry or evolutionary. Due to its importance to epidemiological aspect, the study of geneticvariation of isolates origin from some sources need to be conducted in order to trace the routes of infection.A total of 20 samples obtained from some sources i.e clinically human feces, non-clinically human feces,cattle feces, chicken feces, and beef feces were used in this study. The study was started by confirming allof the isolates using O157 latex agglutination test and H7 antiserum test, followed by genomic DNAanalysis by random amplification of polymorphic DNA /RAPD methods. RAPD results were analyzed using a simple matching coeficient (Ssm) and alogorhythm unweighted pair group method using arithmeticaverages (UPGMA) programe. Results showed there were range of genetic DNA from local isolates (75.1–99,6%) which was almost similar to ATCC 43894 control isolate. The highest similarity (99.6%) to ATCC43894 control was showed by SM-7(1) isolate obtained from cattle fecal and KL-68(1), isolate obtainedfrom clinically human fecal. In addition, KL-52(7) obtained from clinically human fecal had high similarity(99.6%) to MK-35 isolate obtained from chicken fecal. On the other hand, DS-21(4) and DS-16(2) isolatesthat were obtained from beef had high similarity (84.9%) to other isolates including ATCC 43894 controlisolate. The highest similarity of E. coli O157:H7 isolates that were obtained from cattle feces, beef, andchicken feces to human feces isolate indicated that there were both cattle and chicken were potentialreservoirs of the zoonotic agen which can be transmitted to human.
Co-Authors . Sismindari ., Wdjijono Aastuti, Wijayanti Dwi Abdul Salam M. Sofro, Abdul Salam Adi Heru Sutomo Aditya Krishar Karim AETH. Wahyuni, AETH. Afiono Agung Prasetyo Ag. Yuswanto Agnes Sri Harti Agnesia Endamg Tri Hastuti Wahyuni Agnesia Endang Tri Hastuti Wahyuni Agnesia Endang Tri Hastuti Wahyuni Agus Eko Srihanto, Agus Eko Agustinus Joko Nugroho, Agustinus Joko Akiyama, Koichi Al Supartinah Santoso, Al Supartinah Alimuddin . Alimuddin A, Alimuddin Alimuddin, A. Alma Linggar Jonarta, Alma Linggar Ambar Pertiwiningrum Anwar Rosyidi April H Wardhana Ardianata, Dana Arief Mulyono Arum Setiawan Asih Kurniawati B. Sardjono Bambang Hariono Bambang Sumiarto Bambang Sutrisno Banun Kusumawardani Boy M. Bachtiar Budi Mulyaningsih Budi Mulyono Budi Setiadi Daryono BUDI SETIADI DARYONO Charles Rangga Tabbu Charles Rangga Tabbu Chatarina Behar, Chatarina Dayono, Budi Setiadi Dewi Agustina Dewi Seswita Zilda Dito Anggoro, Dito Djaswadi Dasuki Doddi Yudhabuntara Dyah Haryuningtyas Dyah Irnawati Eko Agus Srihanto, Eko Agus Eni Harmayani Enny Yusuf Wachidah Yuniwarti Ety Aryati, Ety Gintung Patantis Gusti Ayu Yuniati Kencana Hardyanto Soebono Hardyanto Subono Hari Eko Irianto Heni Susilowati Heru Susetya Hidayanto, Nur Khusni Hidayati, Dewi Noor I Gusti Ketut Suarjana I Gusti Made Krisna Erawan, I Gusti Made Krisna I Gusti Ngurah Kade Mahardika I Wayan Suardana Ida Tjahajati Ignatius Mulyadi, Ignatius Ika Dewi Ana Ika Dyah Kusumawati, Ika Dyah Indwiani Astuti Istriyati ., Istriyati Istriyati Istriyati Istriyati, . Istriyati, . Istriyati, I. Iwan Dwiprahasto JAKA WIDADA Juni Handajani Karna Wijaya Khilyat Ulin Nur Zaini, Khilyat Ulin Nur Khrisdiana Putri, Khrisdiana LANGKAH SEMBIRING M. Haryadi Wibowo Mammed Sagi Mandojo Rukmo Marsetyawan HNE Soesatyo Marsetyawan HNE. Soesatyo, Marsetyawan HNE. Marsetyawan Soesatyo Masashi Kawaichi, Masashi Maxs Urias Ebenhaizar Sanam Maxs Urias Ebenhaizar Sanam, Maxs Urias Michael Haryadi Wibowo Michael Haryadi Wibowo MM.Firdiana Krisnaningsih Mustofa . Mustofa M, Mustofa Mustofa, M. Ning Rintiswati Nobuyuki Harada Nugroho, Dwi Aji Nugroho, Dwi Aji Nursyirwani . Nuryono ., Nuryono Nuryono, N. Osman Sianipar, Osman Pinandi Sri Pudyani, Pinandi Sri Purnama Edy Santosa Putri, Krisdiana Rahmat Setya Adji Regina TC Tandelilin, Regina TC Regina TC. Tandelilin, Regina TC. Reni Nurjasmi, Reni Rini Widayanti Risya Cilmiaty, Risya S Muharsini S Rahmah Umniyati, S Rahmah Sarwo Edy Wibowo Sebastian Margino Setiyono Setiyono Setyawan Budiharta Sidna Artanto, Sidna Sismindari . Sismindari Sismindari Sismindari, S. Siti Sunarintyas Soemitro Djojowidagdo Sri Darmawati Sri Lestari Sri Murwani Subronto Prodjoharjono Suhartono Taat Putra Surya Amanu Suryani Hutomo, Suryani Susi Iravati Syaiful Anwar Tita Ratya Utari Titik Purwati Widowati, Titik Purwati Tiyas Tono Taufiq, Tiyas Tono Tri Ratnaningsih Tri Untari Tri Wibawa Triyanto . Tsutomu Nohno W. Widodo Wajar, Dony wayan T Artama Wayan T. Artama Wayan T. Artama Wayan Tunas Artama Wayan Tunas Artama Widagdo Sri Nugroho Widagdo Sri Nugroho Widjijono Widjijono Widjijono, W. Widodo . Widya Hary Cahyati Wisnu Nurcahyo Yatri Drastini Yulita Kristanti, Yulita Yuni Wijayanti Yusro Nuri Fawzya ZAKI MUBARAK Zilda, Dewi Zeswita