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Construction of a DNA Vaccine Using Glycoprotein Gene and Its Expression Towards Increasing Survival Rate of KHV-Infected Common Carp (Cyprinus carpio) Nuryati, Sri; Alimuddin, Alimuddin; Sukenda, Sukenda; Soejoedono, Retno Damayanti; Santika, Ayi; Pasaribu, Fachriyan Hasmi; Sumantadinata, Komar
Jurnal Natur Indonesia Vol 13, No 1 (2010)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (120.942 KB) | DOI: 10.31258/jnat.13.1.47-52

Abstract

Deoxyribonucleic acid (DNA) vaccine has recently been developed as an alternative vaccine against virus infection.This study was the first step of DNA vaccine development to protect cyprinids including common carp (Cyprinuscarpio) and fancy koi (Cyprinus carpio) from KHV (koi herpesvirus) infection in Indonesia. One of KHV glycoproteingenes, i.e. glycoprotein (GP) was ligated with Japanese medaka (Oryzias latipes) â-actin promoter to generatepAct/GP as a DNA vaccine. Fourty fish in body weight of 10-15 g/fish were individually injected by pAct/GP intomuscle in different dosage of 2.5 μg, 7.5 μg and 12.5 μg/100 μl phosphate buffer saline. Total RNA was extractedfrom the 12.5 μg of pAct/GP-injected fish muscle at 24, 48 and 67 hours post-injection to analyze GP expression byRT-PCR method. Potential of pAct/GP as DNA vaccine was examined by injecting KHV into the 30-days-vaccinatedfish. Both of possitive and negative control fish group were not vaccinated. Possitive control fish group wereinjected with KHV, but negative control fish group were not. KHV-challenged fish were reared for 1 month, and thedeath fish were calculated daily. Result of RT-PCR analysis showed that GP gene expression were detected at 3 dpost-injection. Expression of GP in the vaccinated fish groups helped to improve their survival rate after challengedby KHV. All of fish without DNA vaccination had dead 17 days after KHV injection. The results demonstrated thatpAct/GP had high potency to be used as a DNA vaccine against KHV infection in cyprinids.
Isolasi dan Identifikasi Aeromonas hydrophila pada Ikan Lele (Clarias gariepinus) Pertambakan Muara Jambi, Provinsi Jambi Wulandari, Titis; Indrawati, Agustin; Pasaribu, Fachriyan Hasmi
Jurnal Medik Veteriner Vol. 2 No. 2 (2019): October
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (465.612 KB) | DOI: 10.20473/jmv.vol2.iss2.2019.89-95

Abstract

Tujuan penelitian ini adalah untuk mengidentifikasi bakteri Aeromonas hydrophila pada ikan lele dengan gejala klinis pada kolam bioflok di Kabupaten Muara Jambi, Provinsi Jambi. Bakteri diisolasi dari 35 ekor ikan lele yang memiliki gejala infeksi A. hydrophila. Pengambilan sampel dilakukan Purposive random sampling. Identifikasi dilakukan uji biokimia berdasarkan Standar Nasional Indonesia SNI 7303.1-2015 dan dilakukan uji konfirmasi metode PCR taget gen aerA. Hasil penelitian menujukkan bahwa sebanyak 35 ekor ikan terisolasi 12 isolat bakteri A. hydrophila pada uji biokimia. Namun pada pengujian PCR pada agarose gel 0.8% menujukkan 8 Isolat posistif gen aerA pada 309 bp. Hasil yang berbeda dapat diakibatkan karena adanya positif palsu pada saat uji biokimia. Hasil identifikasi A. hydrophila dengan metode PCR dapat digunakan hasil yang lebih akurat dan waktu uji lebih cepat dibandingkan dengan uji biokimia. Identifikasi yang cepat dapat mempermudah dalam pencegahan infeksi lebih efektif dan cepat.
Deteksi Gen Penyandi Resistansi blaTEM, blaSHV, dan blaCTXM pada Pseudomonas aeruginosa Ayam Petelur di Kabupaten Cianjur, Jawa Barat Safika, Safika; Arisandi, Fauzan; Pasaribu, Fachriyan Hasmi; Yaddi, Yamin
Jurnal Ilmu dan Teknologi Peternakan Tropis Vol 9, No 1 (2022): JITRO, January
Publisher : Universitas Halu Oleo

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (414.859 KB) | DOI: 10.33772/jitro.v9i1.20448

Abstract

ABSTRAKPseudomonas aeruginosa merupakan bakteri oportunistik patogen yang mampu meninfeksi bagi hewan dan manusia. Resistansi terhadap banyak antibiotik memberikan tantangan yang cukup besar dalam pengobatan infeksi Pseudomonas aeruginosa. Penelitian ini bertujuan mendeteksi adanya resistansi antibiotik dan gen penyandi resistansi pada isolat bakteri Pseudomonas aeruginosa yang diisolasi dari peternakan ayam petelur di Kabupaten Cianjur, Jawa Barat. Sampel diisolasi dan identifikasi sebanyak enam puluh enam melalui usap kloaka. Sampel yang dikoleksi dilakukan kultur pada media selektif (MacConkey agar), dilanjutkan dengan uji mikroskopik, uji biokimia, dan dikonfirmasi dengan secara molekuler dengan polymerase chain reaction (PCR). Sampel yang positif diuji kepekaan terhadap antibiotik menggunakan metode Kirby-Bauer disk diffusion dan mendeteksi gen penyandi resistansi. Hasil penelitian 8 sampel bakteri Pseudomonas aeruginosa dilakukan uji kepekaan antibiotik menunjukkan tingkat resistansi terhadap golongan antibiotik beta laktam (ampisilin 75%) dan aminoglikosida (gentamisin 0%). Dekteksi gen penyandi resistansi secara berturut-turut menunjukkan gen blaTEM (100%), blaCTXM (100%) terdeteksi, sedangkan gen blaSHV tidak terdeteksi pada isolat yang diuji. Perlunya dilakukan penelitian lanjutan untuk mendeteksi sampel dari lingkungan, tempat air minum, pakan maupun karyawan di peternakan tersebut. Sehingga memeberikan informasi dan kajian ilmiah untuk pengaturan regulasi penggunaan antibiotik di peternakan.Kata Kunci: antibiotik, ayam petelur, gen resisten, Pseudomonas aeruginosaDetection of blaTEM, blaSHV, and blaCTXM Resistance Coding Genes in Pseudomonas aeruginosa Layer Chickens in Cianjur Regency, West JavaABSTRACTPseudomonas aeruginosa is a pathogenic opportunistic bacteria capable of infecting animals and humans. Resistance to many antibiotics presents considerable challenges in the treatment of Pseudomonas aeruginosa infections. This study aims to detect the presence of antibiotic resistance and genes encoding resistance in isolates of Pseudomonas aeruginosa isolated from laying hens in Cianjur Regency, West Java. Sixty-six samples were isolated and identified through cloacal swab. The collected samples were cultured on selective media (MacConkey agar), followed by microscopic tests, biochemical tests, and confirmed molecularly by polymerase chain reaction (PCR). Positive samples were tested for susceptibility to antibiotics using the Kirby-Bauer disk diffusion method and detected genes encoding resistance by PCR. The results of the study of 8 samples of Pseudomonas aeruginosa bacteria were tested for antibiotic sensitivity showing the level of resistance to beta-lactam antibiotics (ampicillin 75%) and aminoglycosides (gentamicin 0%). The detection of resistance coding genes, respectively, showed that blaTEM (100%), blaCTXM (100%) genes were detected, while the blaSHV gene was not detected in the tested isolates. Further research is needed to detect samples from the environment, drinking water, feed and employees on the farm. So that it provides information and scientific studies to regulate the regulation of the use of antibiotics in livestock.Keywords: antibiotic, laying hens, Pseudomonas aeruginosa, resistant genes