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Charles Rangga Tabbu
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Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology Veterinary

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Penerapan Metode Diagnosis Cepat Virus Avian Influenza H5N1 dengan Metode Single Step Multiplex RT-PCR Aris Haryanto; Ratna Ermawati; Medania Purwaningrum; Dini Wahyu Yudianingtyas; Michael Haryadi Wibowo; Charles Rangga Tabbu
Jurnal Veteriner Vol 11 No 4 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Avian influenza (AI) virus is a segmented single stranded (ss) RNA virus with negative polarity andbelong to the Orthomyxoviridae family. Diagnose of AI virus can be performed using conventional methodsbut it has low sensitivity and specificity. The objective of the research was to apply rapid, precise, andaccurate diagnostic method for AI virus and also to determine its type and subtype based on the SingleStep Multiplex Reverse Transcriptase-Polymerase Chain Reaction targeting M, H5, and N1 genes. In thismethod M, H5 and NI genes were simultaneously amplified in one PCR tube. The steps of this researchconsist of collecting viral RNAs from 10 different AI samples originated from Maros Disease InvestigationCenter during 2007. DNA Amplification was conducted by Simplex RT-PCR using M primer set. Then, bysingle step multiplex RT-PCR were conducted simultaneously using M, H5 and N1 primers set. The RTPCRproducts were then separated on 1.5% agarose gel, stained by ethidum bromide and visualized underUV transilluminator. Results showed that 8 of 10 RNA virus samples could be amplified by Simplex RTPCRfor M gene which generating a DNA fragment of 276 bp. Amplification using multiplex RT-PCRmethod showed two of 10 samples were AI positive using multiplex RT-PCR, three DNA fragments weregenerated consisting of 276 bp for M gene, 189 bp for H5 gene, and 131 bp for N1. In this study, rapid andeffective diagnosis method for AI virus can be conducted by using simultaneous Single Step Multiplex RTPCR.By this technique type and subtype of AI virus, can also be determined, especially H5N1.
Akumulasi Timbal dalam Cakar Ayam Kampung Djohan -; Charles Rangga Tabbu
Jurnal Veteriner Vol 11 No 1 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Domestic chickens tend to consume lead (Pb) from their contaminated environment especially in freerangingchickens. Feet of domestic chickens are commonly consumed by many people, making them goodbioindicator to be analyzed for environmental monitoring and food safety purposes. Accumulation of leadin feet and parts of feet (tarsometatarsus bones, digiti, and skin-muscles) were investigated in this study.The average concentrations of lead in whole feet, tarsometatarsal bones, digiti, and skin-muscles were 3.4,3.8, 3.3, and 1.9 ?g.g-1 d.m., respectively. The average amount of Pb accumulated in chicken feet ranges from25.5 to 74.7 ?g. Consumption of 0.5 – 2 chicken feet. A day-1 (low – high intake) results in the exposure of 4,8to 111,5 ?g. individual-1.day-1, which was much higher than the daily exposure standar of 1 ?g. individual-1. day-1. However, very low intake (< 0.1 chicken feet.day-1) results in exposure lower than the recommendedexposure standard. More frequent monitoring and exposure assessment combined with the awareness ofgeneral public on lead contamination in the environment are important for minimizing the risk of leadexposure to human through chicken feet consumption.
Deteksi Bovine Herpesvirus-1 Secara Immunohistokimia pada Membran Korioallantois Telur Ayam Berembrio (IMMUNOHISTOCHEMISTRY DETECTION OF BOVINE HERPESVIRUS-1 IN CORIOALLANTOIC MEMBRANE OF CHICKEN EMBRYONATED EGG) Yuli Purwandari Kristianingrum; Charles Rangga Tabbu; Bambang Sutrisno; Sitarina Widyarini; Kurniasih .; Tri Untari; Asmarani Kusumawati
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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Abstract

Infectious Bovine Rhinotracheitis (IBR) is caused by Bovine Herpes virus-1 in the cattle. The clinicalsigns demonstrate depression, anorexia, swelling of the vulva, redness of the vestibule, pustule and ulceron the vaginal mucosal. Based on previous research, IBR virus from the nasal swab could be grown inchorio-allantoic membrane of embryonated chicken eggs. This study aim was to confirm whether IBR virusin cattle could be grown in embryonated chicken eggs as a substitute for cell culture. A total of five nasalswab samples from the cows that were positive for IBR infection (diagnosed by Polymerase Chain Reactionand cell culture) were inoculated on the chorio-allantois membrane of embryonated chicken eggs.Observation of lesions performed at 3-5 days after inoculation. Re-inoculation (passage) was done threetimes. Pock characteristic lesions were observed on the corioallantoic membrane with the size of 5-7 mm,rounded shape, opaque edge, with necrosis in the central area. Furthermore, pock lesions were processedfor hematoxylin and eosin staining and immuno-histochemistry. The result of hematoxylin and eosinstaining showed that the formation of intranuclear inclusion bodies and vacuolization of the epithelial cellof membrane was observed. Immuno-histochemistry staining showed positive reaction for antibodiesagainst BHV-1 in the epithelial cells membrane. In conclusion, embryonated chicken eggs could be usedas a medium for detection of IBR.
Non Coding Region dan Amino Terminus Gen Polimerase Asidik Virus Avian Influenza Subtipe H5N1 Asal Hewan Indonesia Gusti Ayu Yuniati Kencana; Widya Asmara; Charles Rangga Tabbu; I Gusti Ngurah Kade Mahardika
Jurnal Veteriner Vol 11 No 3 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

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.
VARIATION OF NON-CODING REGION AND CODING REGION OF 5’-TERMINAL CRNA OF POLYMERASE BASIC 1 OF AVIAN INFLUENZA VIRUS SUBTYPE H5N1 Gusti Ayu Yuniati Kencana; Widya Asmara; Charles Rangga Tabbu; I Gusti Ngurah Kade Mahardika
Jurnal Veteriner Vol 10 No 1 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

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
Akumulasi Timah Hitam dalam Daging dan Tulang Ayam Kampung dan Ayam Negeri (LEAD ACCUMULATION IN MEAT AND BONES OF DOMESTIC AND BROILER CHICKEN) Djohan .; Charles Rangga Tabbu
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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Abstract

Lead is a heavy metal polluting the environment, and its accumulation in animal or human bodies canhave neurotoxic and nephrotoxic effects on animals and human. Lead-contaminated chicken meat can bethe source of lead to human. Lead exposure to human can be assessed by measuring its concentration andaccumulation in chicken body parts and analyzing chicken consumption patterns. This study was conductedto measure lead concentrations in chicken body parts and to estimate lead exposure caused by consumptionof chicken body parts (breast, legs, wings) and tissues (meat, skin, cartilage, spongy bones). Samples wereextracted by using aqua regia digestible method with a mixture of HCl: HNO3 (3:1; v/v) and leadconcentrations were measured by the Atomic Absorption Spectrophotometry (AAS) method. The leadconcentrations in chicken tissues varied from< 0.01 to 1.81?g.g-1dry weight.The average concentrations oflead in chicken tissues were lower than the recommended safety level of lead in chicken meat (1.0?g.g-1),except for the breast cartilage (1.03?g.g-1). The lowet accumulation level 2.6 ?g g-1 was found in domesticchicken wings while the highest of 32.9 ?g g-1 was found in broiler chicken breast (total of meat, skin,cartilage). Based on the data of lead accumulation in chicken tissues, a polynomial equation describing theprobability (P) to be exposed to certain amount of lead in chicken tissues (A, in ?g) was determined as P =-(1 x 10-3)A2 + (6,4 x 10-2)A.
Co-Authors Agnesia Endang Tri Hastuti Wahyuni Agus Eko Srihanto Aris Haryanto Aris Purwantoro Asmarani Kusumawati Bambang Sumiarto Bambang Sutrisno Dini Wahyu Yudianingtyas Djohan - Djohan . Dyah Ayu Widiasih Enny Yusuf Wachidah Yuniwarti Gusti Ayu Yuniati Kencana Haryadi M. Wibowo Heru Susetya Heru Susetyo I Gusti Ngurah Kade Mahardika Khrisdiana Putri Kurniasih . M. Haryadi Wibowo Medania Purwaningrum Michael Haryadi Wibowo Ratna Ermawati Rini Widayanti Setyawan Budiharta Sidna Artanto Sitarina Widyarini Tati Aryani Tri Untari Vinsa Cantya Prakasita Wayan Tunas Artama Widya Asmara Yuli Purwandari Kristiangingrum Yuriadi -