AEVI-10 Investigasi Outbreak MCF di Kabupaten Musi Rawas Tahun 2016

Malignant Catarrhal Fever (MCF) merupakan penyakit viral yang menyerang sapi, rusa, bison, kerbau dan ruminansia lainnya. Sapi muda biasanya lebih rentan terutama yang dipelihara bersama domba (Damayanti, 2005). Penyakit ini kebanyakan berakibat dengan kematian (Teankam et al., 2000; Schultheiss, et al., 2000; Barker et al., 1993). MCF disebabkan oleh alcelaphine herpesvirus 1 (AlHV-1) dan ovine herpesvirus 2 (OvHV-2) (Fenner et al., 2011). Penyakit bersifat sporadik dengan tingkat  kematian dapat mencapai  100%  (Hamilton,  1990),  meskipun ada hewan yang sembuh setelah terserang MCF (Penny, 1998). Penyakit  MCF di  Indonesia dilaporkan pertama  kali  oleh  Paszotta  pada  tahun  1894  di Kediri,  Jawa  Timur  (Mansjoer,  (1954)  dalam Khatimah, dkk. 2014).  Penyakit  MCF telah  tersebar  hampir  di seluruh  Indonesia.  Namun di beberapa  daerah  banyak  kejadian  MCF  tidak terdiagnosis  atau  tidak dilaporkan.  Kerugian yang  ditimbulkan  akibat  penyakit  MCF  cukup besar  terutama  jika  kasus  penyakit  ini  terjadi pada hewan bibit. Kejadian penyakit MCF pada sapi bali di lapangan  sering  dikaitkan  dengan adanya ternak domba (Partadiredja et al., 1988). Lesi sapi penderita MCF dapat dilihat dari  gambaran  patologi anatomi dan histopatologi yang patognomonik  yaitu  vaskulitis  berupa  infiltrasi limfosit, makrofag, neutrofil  dan  sel  plasma pada  beberapa  organ seperti otak, paru-paru, hati, jantung dan usus (Liggit dan De Martini, 1980).  Kegiatan penyidikan kematian sapi bali  yang diduga disebabkan oleh Malignant Catarrhal Fever virus oleh tim Balai Veteriner Lampung di Kabupaten Musi Rawas dilaksanakan berdasarkan permohonan investigasi oleh Dinas Peternakan Kabupaten  Musi Rawas  mengenai adanya laporan kasus kematian sapi bali dengan gejala klinis mengarah pada penyakit  MCF di desa L Sidoharjo Kecamatan Tugu mulyo. Berdasarkan laporan tersebut maka Balai Veteriner mengeluarkan Surat Perintah Tugas No. 16001/TU.040/F5.C/09.2016 untuk melakukan penyidikan bersama dinas Peternakan Kabupaten Musi Rawas.Tujuan kegiatan adalah melakukan penyidikan kejadian kematian sapi bali di Kabupaten Musi Rawas, melakukan pengumpulan data epidemiologis, pengambilan spesimen di lapangan untuk mengetahui penyebab kematian sapi bali di Kabupaten Musi Rawas

ANALISIS MOLEKULER FILOGENETIK DAN STRUKTUR ANTIGENIC VIRUS AVIAN INFLUENZA SUBTIPE H5N1 ISOLAT LAMPUNG TAHUN 2008-2013

Penelitian ini bertujuan melakukan karakterisasi molekuler antigenic site terhadap isolat virus avian influenza (AI) Balai Penyidikan dan Pengujian Veteriner (BPPV) Regional III Lampung dari tahun 2008-2013. Amplifikasi RNA dilakukan dengan teknik reverse transcription polymerase chain reaction (RT-PCR) menggunakan 4 pasang primer referens dari Australian Animal Health Laboratory (AAHL) Geelong Australia (HA10, HA20, dan HA30) dan dilanjutkan dengan proses pengurutan. Analisis hasil pengurutan dengan menggunakan perangkat lunak MEGA versi 5.05 yang meliputi multiple alignment, deductive amino acids prediction, dan phylogenic tree analysis diperoleh hasil perbedaan genetik antar isolat Lampung dari tahun 2003-2013 ditemukan berkisar 1,1-9,1% dengan tingkat homologi mencapai 90,9-98,9%. Variasi genetik ditemukan adanya substitusi pada posisi 53 (R53K), 126 (D126E), 136 (P136), 138 (H138Q, dan H138L), 140 (R140K, R140S, dan R140N), 141 (S141P), dan 189 (K189R). Berdasarkan analisis filogenic tree isolat Lampung tahun 2008-2011 termasuk ke dalam clade 2.1.3. Analisis filogenik isolat AI tahun 2012-2013 yang menginfeksi unggas air mempunyai homologi sekitar 98,5-99,1% dibandingkan dengan isolat AI yang menginfeksi unggas air asal Jawa dan termasuk ke dalam clade 2.3.2.1

Comparative lactic acid bacteria (LAB) profiles during dadih fermentation with spontaneous and back-slopping methods, as identified by terminal-restriction fragment length polymorphism (T-RFLP)

The diversity of lactic acid bacteria (LAB) present during the manufacture of traditional fermented buffalo milk from West Sumatra, known as dadih, was studied via a culture-independent approach using terminal-restriction fragment length polymorphism (T-RFLP) to compare the dynamic diversity in back-slopping and spontaneous fermentation methods. Total LAB and pH were measured in freshly prepared buffalo milk and in \textit{dadih} fermented for 24 and 48 hours. The results indicated significant differences between the fermentation methods, with higher total LAB, and greater phylotype richness and relative abundance being identified in the back-slopping method. Terminal fragment lengths (TRFs) of 68 and 310 bp were common to both techniques, similar to those of Lactobacillus fermentum, Fructobacillus pseudoficulneus, Leuconostoc citreum, Leuconostoc kimchii, and Leuconostoc sp. The changes in phylotype number (species number) and relative abundances of LAB communities identified are expected to produce data needed to formulate the best fermentation process for dadih manufacturing. A 24-hour back-slopping fermentation method is recommended, as fermentation time of longer than 24 hours reduced viable LAB significantly. Our results also indicated that the T-RFLP technique is not only clearly sensitive enough and adequate for segregating LAB diversity in both fermentation methods, but that it also provides good information regarding the structure of microbial communities and their composition change during the fermentation process

The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia

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

Characterization of Newcastle Disease Virus Isolated from Peacocks in Palembang City, South Sumatra

Introduction: Newcastle Disease (ND) is an infectious disease in various types of poultry caused by the Newcastle Disease Virus (NDV). Cases of ND in Indonesia have been reported in commercial and backyard chickens, pigeons, ducks and geese, even in eagles and peacocks. Peacock is a wild bird protected by Indonesia's Government Regulation No. 7 of 1999. This study aims to identify, isolate and characterize the NDV molecularly in cases diagnosed as ND in peacocks. Method: Samples were obtained from organs (lungs and spleen) of peacocks which showed neurological symptoms, diarrhoea and then died. Real-time RT-PCR ND was used to identify the cause of death of the peacock. Virus isolation and observation of embryonic changes and death were carried out on embryonic chicken eggs. Sequencing was carried out to characterize the F and HN entire genes of the NDV. The nucleotide sequences were analyzed using MEGA-X software, including amino acid prediction, analysis of genetic variation at the amino acid level, homology and construction of the phylogenic tree. Result: The results of the sample identification were positive for the Newcastle disease virus. Observations of chicken embryos are stunted, have few feathers, are haemorrhagic, and die in less than 60 hours. Virus isolation was obtained with a titer of 26. Molecular analysis showed that the RRQKRF cleavage site pattern in the F gene had homology of 95.8-97.6% and was in the same branching area as the previous ND virus in Indonesia. There were no amino acid mutations at the antigenic site, glycolysis and neutralization epitopes in the HN gene. Conclusion: The virus isolated from the peacock is a velogenic strain of NDV, subgenotype VII.2 and has a close genetic range to the NDV that has been previously reported in commercial and domestic poultry. This result shows that ND is also a threat to protected wild birds

The Development of Pathogenicity of Avian Influenza Virus Isolated from Indonesia

Highly pathogenic avian infl uenza outbreak in Indonesia has been reported in various poultry due toH5N1 subtype. The presence of multiple basic amino acids within the cleavage site of HA glycoprotein hasbeen identifi ed to be associated with the pathogenicity of avian infl uenza virus. The study was retrospectivestudy which was designed to characterize the cleavage site and fusion site region of haemagglutinin gene ofAIV isolated from various poultry in 2003 to 2013. Isolation, Identifi cation and propagation were carried outto collect viral stock. For virus detection, reverse transcriptase PCR (RT-PCR) method on H5 and N1 genefragment was performed. All of RT-PCR HA gene positive products were sequenced for further nucleotideanalysis and to determine the nucleotide composition at the targeted fragment. The results are all AIV isolateswere identifi ed as H5N1 subtype. The sequence analyses revealed some motives of basic amino acid motivethat were classifi ed as highly pathogenic avian infl uenza virus. Further analyses on fusion domain of all AIVisolated during the period 2003 to 2013 showed conserved amino acid.Keywords: avian infl uenza, haemagglutinin, cleavage site, basic amino acid, fusion site</div

The Development of Pathogenicity of Avian Influenza Virus Isolated from Indonesia

Highly pathogenic avian infl uenza outbreak in Indonesia has been reported in various poultry due toH5N1 subtype. The presence of multiple basic amino acids within the cleavage site of HA glycoprotein hasbeen identifi ed to be associated with the pathogenicity of avian infl uenza virus. The study was retrospectivestudy which was designed to characterize the cleavage site and fusion site region of haemagglutinin gene ofAIV isolated from various poultry in 2003 to 2013. Isolation, Identifi cation and propagation were carried outto collect viral stock. For virus detection, reverse transcriptase PCR (RT-PCR) method on H5 and N1 genefragment was performed. All of RT-PCR HA gene positive products were sequenced for further nucleotideanalysis and to determine the nucleotide composition at the targeted fragment. The results are all AIV isolateswere identifi ed as H5N1 subtype. The sequence analyses revealed some motives of basic amino acid motivethat were classifi ed as highly pathogenic avian infl uenza virus. Further analyses on fusion domain of all AIVisolated during the period 2003 to 2013 showed conserved amino acid. Keywords: avian infl uenza, haemagglutinin, cleavage site, basic amino acid, fusion sit

Qualitative Analysis of Partial 16S rRNA Amplicon of Mitochondrial Gene of Stingless Bees in Pesawaran

Pesawaran is one of the areas that has the potential to cultivate stingless bees as producers of honey and propolis. This study was aimed at qualitatively analysing partial amplicons of the 16S rRNA gene from individuals found at the research site. The research was performed from April to September 2023 in an explorative approach. Based on the exploration that has been done in Harapan Jaya Village, Way Ratai District, Pesawaran Regency, individual stingless bees were obtained from 5 different colonies. Individual samples of stingless bees were further extracted and amplified by commercial kits. Each individual from 5 different colonies has successfully obtained its DNA amplicons. Based on qualitative analysis using 1% agarose gel, the size of the partial gene ranged from 400bp to 500bp. This result is in accordance with the data contained in the genbank, where the 16S rRNA gene size is more than 500bp. Therefore, it has been concluded that the size of the partial amplicons of the 16S rRNA gene of individual stingless bees from the 5 colonies that were obtained is actually in range of the size of the 16S rRNA gene of stingless bees that have been recorded in the genbank

ANALISIS GENETIK GEN HEMAGLUTININ VIRUS AVIAN INFLUENZA SUBTIPE H5N1 ISOLAT LAMPUNG TAHUN 2008-2013

Avian influenza is an infectious poultry disease which be caused by type A Avian Influenza virus in the family of Orthomyxoviridae. AI outbreaks in Lampung was reported in 2003 in South Lampung district which continued to spread in several districts/cities in the region of DIC Region III. Molecular data of the Lampung isolates has not been reported. This study was aimed to characterize the full HA gene. AI virus isolates derived from the work area of DIC Region III were isolated on chicken embrionated eggs negative antibody spesific (SAN) to AI at 10 days of age. Allantois fluid was extracted to obtain RNA and the subtype identification of H5N1 was done by Reverse Transcription Polymerase Chain Reaction (RT-PCR) technique. Amplification of the HA gene using 4 pairs references primer from the Australian Animal Health Laboratory (AAHL) Geelong Australia (HA10, HA20, HA30 and HA40) and was continued with the sequencing process. Sequences were analysed by the MEGA software version 5.05. Analysis of genetic differences between isolat Lampung from 2003-2008 compare with isolat from 2008-2013 indicated genetic differences among 1.1- 9.1% and homology among this isolat were 90.9-98.9%. Genetic variation found in the antigenic site and the receptor binding site with a substitution at position 53 (R53K), 126 (D126E), 136 (P136S), 138 (H138Q and H138L), 140 (R140K, R140S and R140N), 141 (S141P), 189 (K189R), 129 (S129L) and 189 (K189R). Analysis of the composition of amino acids at cleavage site of Lampung isolates was classified to HPVAI. The feature of sequences were QRERRRKKRG, QRESRRKKRG and QRERRRKG. Phylogenic tree analysis of tree Lampung in 2008-2011 isolates belong to the clade 2.1.3. Analysis of the 2012-2013 AI isolates that infect waterfowl have approximately 98.5-99.1% homology compared with AI isolates that infect waterfowl Javanese origin and belong to the clade 2.3.2.2

Diversity of lactic acid bacteria in dadih produced by either back-slopping or spontaneous fermentation from two different regions of West Sumatra, Indonesia

Aim: Dadih samples from two different origins (Kamang and Gadut in West Sumatra) manufactured with different methods (back-slopping or spontaneous fermentation) were evaluated for the diversity of lactic acid bacteria (LAB). Materials and Methods: Four dadih samples manufactured with two different fermentation methods were obtained from Kamang and Gadut regions. Both genotypic and phenotypic characteristic (16S rRNA partial gene sequence analysis and carbohydrate fermentation profile) were used to analyze the diversity of dadih LAB population. Results: This study showed that LAB count in back-slopping fermented dadih was one log cycle higher than spontaneous fermented dadih. LAB isolates from the two regions were divided into three genera, namely Lactococcus, Lactobacillus, and Pediococcus. Sequencing results showed that 41.6% (five isolates) were identified as Lactococcus lactis ssp. lactis, 25% (three isolates) were identified as Lactobacillus plantarum ssp. plantarum, 16.6% (two isolates) were identified as L. lactis ssp. cremoris, and 8.3% (one isolate each) were identified as Pediococcus pentosaceus and Lactobacillus pentosus. Conclusion: Five species were determined in back-slopping fermented dadih, i.e., L. lactis ssp. lactis, L. lactis ssp. cremoris, L. plantarum ssp. plantarum, L. pentosus, and P. pentosaceus. On the other hand, spontaneous fermented dadih only contained three different species, namely L. lactis ssp. lactis, L. lactis ssp. cremoris, and L. plantarum ssp. plantarum. This research showed that back-slopping fermentation offers greater abundance and diversity compared to spontaneous fermentation in dadih