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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 : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

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.
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 : The Faculty of Veterinary Medicine, Udayana University

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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.
HISTOPATOLOGI IKAN KERAPU MACAN YANG DIIMBUHI BAKTERI ASAM LAKTAT DAN DIUJI TANTANG VIBRIO ALGINOLYTICUS (HISTOPHATOLOGY OF TIGER GROUPER SUPPLEMENTED WITH LACTIC ACID BACTERIA AND CHALLENGED BY VIBRIO ALGINOLYTICUS) ., Nursyirwani; Asmara, Widya; Wahyuni, Agnesia Endang Tri Hastuti; ., Triyanto
Jurnal Veteriner Vol 16 No 4 (2015)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Supplementation of lactic acid bacteria (LAB) as probiotic in aquaculture has been reported toincrease fish growth and enhance their resistance against diseases. The aim of this study was to figureout histological changes of tiger grouper (Epinephelus fuscoguttatus) fed with LAB isolates followed bycha
KARAKTERISASI GEN NON STRUKTURAL 1 (NSI) VIRUS AVIAN INFLUENZA PADA ISOLAT ITIK TAHUN 2013 Hidayanto, Nur Khusni; Asmara, Widya; Wibowo, Michael Haryadi
Jurnal Sain Veteriner Vol 33, No 2 (2015): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (770.282 KB) | DOI: 10.22146/jsv.17919

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Wabah avian influenza (AI) di Indonesia telah terjadi sejak akhir tahun 2003 dan masih terjadi secara endemis sampai sekarang. Pada akhir tahun 2012 terjadi kasus kematian yang cukup tinggi pada itik yang disebabkan penyakit AI subtipe H5N1 yang diklasifikasikan ke dalam clade 2.3.2, sedangkan virus AI sebelumnya diklasifikasikan pada clade 2.1.1, 2.1.2 dan 2.1.3. Salah satu yang berperan dalam virulensi penyakit AI adalah motif asam amino C-terminal protein nonstruktural1 (NS1). Analisis sekuens virus influenza terutama protein nonstruktural 1 (NS1) yang berasal dari unggas mempunyai motif asam amino ESEV pada Cterminal sedang pada virus influenza manusia mempunyai motif RSKV pada C-terminal. Data sekuen NS1 untuk virus AI terbaru belum lengkap sehingga perlu disekuen untuk melengkapi data molekuler NS1 virus AI.Residu C-terminal protein NS1 virus avian influenza subtype H5N1 perlu dikaji karena mempengaruhi patogenisitas dan virulensi virus. Penelitian ini bertujuan untuk mengetahui motif asam amino pada C-terminal protein nonstruktural 1 (NS1) virus avian influenza yang menyerang itik pada tahun 2013. Sampel berasal dari kasus AI pada itik di Tulungagung dan Blitar pada tahun 2013. Isolasi virus menggunakan telur berembrio tertunas ayam. Identifikasi virus AI subtipe H5N1 menggunakan teknik reverse transcription polimerase chain reaction (RT-PCR) dengan primer H5 (Lee et al., 2001) dengan target amplifikasi 545 bp dan primer N1 (Payungporn et al., 2004) dengan target amplifikasi 131 bp. Amplifikasi gen NS1 menggunakan RT-PCR dengan 2 pasang primer (Bannet-Noah et al., 2007) yang didesain mengamplifikasi gen NS dan dilanjutkan proses sekuensing gen NS1. Sekuen yang diperoleh dianalisa dengan menggunakan software MEGA 5.05 yang meliputi multiple alignment, prediksi asam amino dan analisis pohon filogenetik. Hasil sekuen isolat diperoleh panjang nukleotida yang mengkode protein NS1 sepanjang 690 nt. Hasil analisis pohon filogenetik menunjukkan bahwa ke lima isolat uji tidak berada satu grup dengan dengan isolat asal Indonesia tahun 2003- 2008 dan berdekatan dengan klaster virus AI yang berasal dari Asia clade 2.3.2. Pada semua isolat tahun 2013 ditemukan delesi asam amino pada posisi 80-84, substitusi asam amino D92E, asam amino 149 semua isolat mempunyai asam amino alanine (A), asam amino ke 196 ditemukan adanya variasi substitusi berupa lysine (K) dan glutamic acid (E) dan asam amino pada gen NS1 mempunyai motif ESEV pada posisi PDZ ligand.
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 : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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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.
EVALUASI STATUS VIRULENSI ISOLAT BACILLUS ANTHRACIS ASALNUSA TENGGARA DAN PAPUA MENGGUNAKAN METODE POLYMERASE CHAIN REACTION MULTIPLEX Ebenhaizar Sanam, Maxs Urias; Asmara, Widya; Tri Hastuti Wahyuni, Agnesia Endang; Wibowo, Michael Haryadi; Adji, Rahmat Setya
Jurnal Kedokteran Hewan Vol 9, No 2 (2015): J. Ked. Hewan
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v9i2.2802

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Polyphenols extracted from the Green Tea (Camellia sinensis) augments the protective immune responses in mice challanged with Salmonella typhimurium Ratnaningsih, Tri; Asmara, Widya; Sismindari, Sismindari
Medical Journal of Indonesia Vol 13, No 1 (2004): January-March
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (333.117 KB) | DOI: 10.13181/mji.v13i1.122

Abstract

Green tea is an aqueous infusion of dried unfermented leaves of Camellia sinensis. Numerous biological activities of green tea have been reported. The aqueous infusion and its polyphenolic substance are known for their activity as an antimutagenic,  antibacterial, hypocholesterolemic, antioxidant, and mutagenic of B lymphocyte. Studies have demonstrated that green tea polyphenols increase IL-12 production. Salmonella spp infection is an important public health problem in many countries. Cell-mediated immunity (CMI), especially T-cell help is important for protection against this infection. Recent evidence indicates that IL-12 is one such factor that plays a crucial role in the development of CMI. These studies were carired out to investigate the effect of green tea polyphenols to the immune cellulare in mice responses of mice during Salmonella typhimurium infection. The subject consisted of 36 female mice (Balb/C), 6-8 weeks old, divided into 3 groups. The first group was given 10 mg polyphenols/mouse, the second group was given 5 mg polyphenols/mouse, and the third group as the control. In day 31, all mice were infected with 108 CFU Salmonella typhimurium orally. On day 0, 3, 5, and 7 postinfection, 3 mice from each groups were sacrificed, the splenocytes were extracted and cultured to measure  the level of IFN-g in the supernatan and. The peritoneal macrophages were also extracted and cultured to measure the phagocytic activity. The level of IFN-g in splenocyte culture supernatant  increased during infection  in all groups, but the level of the experimental groups  were higher than in control group. The  percentage of phagocytic activity of peritoneal macrophages were higher in the experimental groups than in the control group. The increase of the phagocytic activities were seen corelate with the level of IFN-g supernatan splenocyte culture. (Med J Indones 2004; 13: 1-7)Keywords: polyphenols, green tea, macrophages, phagocytosis
EVALUASI KIT DETEKSI CEPAT TERHADAP SAMPEL OTAK ANJING TERINFEKSI VIRUS RABIES Wibowo, Michael Haryadi; Untari, Tri; Artanto, Sidna; Amanu, Surya; Wahyuni, AETH.; Asmara, Widya
Jurnal Kedokteran Hewan Vol 9, No 1 (2015): J. Ked. Hewan
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v9i1.2797

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Penelitian ini bertujuan mengetahui potensi kit deteksi cepat Anigen® rapid test kit rabies Ag dalam mendeteksi virus rabies pada sampel otakanjing yang diperoleh dari lapangan yang meliputi batas deteksi, kecepatan reaksi, uji reaksi silang, uji sensitivitas, dan spesifisitas. Batas deteksi ditentukan dengan pengenceran secara serial kontrol positif virus rabies dan selanjutnya diuji dengan rapid test kit sesuai petunjuk produsen. Uji reaksi silang dilakukan dengan canine parvovirus, Escherichia coli, dan Salmonella sp. Uji sensitivitas dan spesifisitas dilakukan terhadap sampel otak yang telah dikonfirmasi positif rabies dengan uji fluorescent antibody technique. Konfirmasi uji rapid test tersebut dilakukan dengan reverse transcriptase polymerase chain reaction. Berdasarkan data yang diperoleh dalam penelitian ini dapat disimpulkan bahwa Anigen® rapid test kit rabies Ag mampu mendeteksi sampel yang mengandung virus rabies dengan titer 0,5 x log 106,5/0,03 ml, dengan rata-rata kecepatan reaksi 1,8 menit 29,35 detik (kurang dari 2 menit). Di samping itu Anigen® rapid test kit rabies menunjukkan tidak terdapat reaksi silang dengan canine parvovirus, Escherichia coli, dan Salmonella sp. serta mempunyai sensitivitas 92,30% dan spesifisitas 97,90%
Placental Trophoblast Responses to Porphyromonas gingivalis Mediated by Toll-like Receptor-2 and -4 Kusumawardani, Banun; Soesatyo, Marsetyawan HNE; Dasuki, Djaswadi; Asmara, Widya
Journal of Dentistry Indonesia Vol 20, No 2 (2013): August
Publisher : Faculty of Dentistry, University of Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (935.461 KB) | DOI: 10.14693/jdi.v20i2.150

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Trophoblast participates in preventing allorecognition and controlling pathogens that compromise fetal wellbeing. Toll-like receptors recognize conserved sequences on the pathogens surface and trigger effector cell functions. Porphyromonas gingivalis is thought to spread to the umbilical cord and cause fetal growth restriction. Objective: To characterize expression and function of TLR-2 and TLR-4 in trophoblast cells from Porphyromonas gingivalisinfected pregnant rats. Methods: Live Porphyromonas gingivalis were challenged into the maxillary first molar subgingival sulcus of female rats before and/or during pregnancy and sacrified on gestational day (GD) 14 and 20. Porphyromonas gingivalis was detected by API-ZYM system in the maternal blood of the retro-orbital venous plexus and the umbilical cord. TLR-2 and TLR-4 expressions in trophoblast cells was detected by immunohistochemistry. Results: Porphyromonas gingivalis was first detected in the maternal blood and finally spread to the umbilical cord. Syncytiotrophoblast, spongitrophoblast and trophoblastic giant cell in treated groups had significantly higher expression of TLR-2 and TLR-4 than control group (p<0.05). Conclusion: Syncytiotrophoblast, spongitrophoblast and trophoblastic giant cell are able to recognize Porphyromonas gingivalis through TLR-2 and TLR-4 expression. The ligation of TLR-2 and TLR-4 promoted cytokine production and induced trophoblast cell death. These findings strengthen links between periodontal disease and fetal growth restriction.DOI: 10.14693/jdi.v20i2.150
Gestational Day-Dependent Expression of Interleukin-10 and Tumor Necrosis Factor-alpha in Porphyromonas gingivalis-infected Pregnant Rats Kusumawardani, Banun; Soesatyo, Marsetyawan HNE; Dasuki, Djaswadi; Asmara, Widya
Journal of Dentistry Indonesia Vol 20, No 3 (2013): December
Publisher : Faculty of Dentistry, University of Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (926.193 KB) | DOI: 10.14693/jdi.v20i3.199

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Fetal growth restriction remains a major cause of neonatal morbidity and mortality. Porphyromonas gingivaliscan induce placental inflammatory response resulting in fetal growth restriction. Objective: This study aimed to evaluate the potential utility of the pro-inflammatory cytokine TNF-α and anti-inflammatory cytokine IL-10 in rat placental tissues to understand whether these events were causally related. Methods: Female rats were infected with live-Porphyromonas gingivalis at concentration of 2x109 cells/ml into subgingival sulcus area of the maxillary first molar before and/or during pregnancy. They were sacrificed on gestational day (GD)-14 and GD20. The expression of TNF-α and IL-10 in macrophages and trophoblast cells were detected by immunohistochemistry. Results: A higher expression of TNF-α was found in spongiotrophoblast of the Pg-BD group on GD14 (6.30±1.16), and in trophoblastic giant cells of Pg-D group on GD20 (5.50±1.35). Furthermore, a higher expression of IL-10 was found in trophoblastic giant cells of the Pg-BD group on GD14 (4.50±1.51) and in syncytiotrophoblasts of Pg-BD group on GD20 (8.70±2.67). Conclusion: The expression of TNF-α on GD14 and GD20 were accompanied by increased expression of IL-10. The placental pathologic conditions induced by Porphyromonas gingivalis can be inhibited by elevated expression of IL-10 in macrophages and trophoblast cells.DOI: 10.14693/jdi.v20i3.199
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 Agnes Sri Harti 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 April H Wardhana Ardianata, Dana Arum Setiawan Asih Kurniawati B. Sardjono Bambang Hariono Bambang Sumiarto Bambang Sutrisno Banun Kusumawardani Boy M. Bachtiar Budi Mulyaningsih Budi Mulyono 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 Dyah Haryuningtyas Dyah Irnawati Eko Agus Srihanto, Eko Agus Eni Harmayani Ety Aryati, Ety Gintung Patantis Gusti Ayu Yuniati Kencana Hardyanto Soebono Hardyanto Subono Hari Eko Irianto Heni Susilowati Hidayanto, Nur Khusni Hidayati, Dewi Noor I G. Made Krisna Erawan 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