Abstract. All cattle in 20 dairy herds randomly selected from herds participating in the Dairy Herd Improvement Association program in 2 counties in central Michigan were tested for the presence of bovine viral diarrhea virus (BVDV). Virus-positive animals were retested to ascertain persistent infection with the virus. A total of 5,481 animals were tested for presence of BVDV. In 9 of the herds, all animals were also tested for virus neutralizing antibody titer. Based on infection and vaccination status, these 9 herds were divided into 3 different herd categories: A, 5 herds with currently no cattle persistently infected (PI) with BVDV and without any vaccination program against BVDV in recent years; B, 2 herds with no current PI cattle but using killed BVDV vaccines; and C, 2 herds with PI cattle. PI cattle were detected in 3 out of 20 herds (15%). A total of 7 of 5,481 animals (0.13%) were PI. The mean prevalences of antibody carriers in herd categories A, B, and C were 28.8%, 76.4% and 90.6%, respectively. For one herd in category A, antibody analyses indicated that mostly young stock was seropositive, suggested recent BVDV infection in a previously closed and naive herd. Cattle in category B herds were vaccinated with killed vaccine from the age of 15 months. These herds had several antibody negative animals among the younger cows, suggesting incomplete protection against BVDV infection. In the 3 herds in which PI animals were detected, all cattle had been vaccinated with killed vaccine. The antibody-positive animals had antibody titers that were significantly different both among herds and among herd categories. The antibody titers of animals exposed to PI animals were significantly higher than those of animals vaccinated with killed vaccine.

Background and Aim: A previous study divided Indonesian bovine viral diarrhea virus (BVDV)-1 into subgenotypes BVDV-1a to BVDV-1d based on the partial NS5B gene using strain Bega as reference for BVDV-1a. In fact, it is clustered into BVDV-1c with strain Bega-like Australia. BVDV genotyping has been done on isolates from Jakarta, West and Central Java, but East Java isolates have not been genotyped. This study aimed to analyze genetic variability and amino acid residues in the nucleotide-binding pocket of the NS5B gene from infected cattle. Materials and Methods: Samples were obtained from the Sera Bank originating from active and passive surveillance of cattle that had been tested for BVDV antigen from 2013 to 2017. Detection of the p80 antibody and BVDV genotyping was carried out using ELISA and nested-multiplex-polymerase chain reaction (PCR), respectively. We defined 15 nested PCR products for partial sequencing of NS5B. Those field samples were selected from each location and year using proportional calculation as a representative sample. Homological and phylogenetic analyses of the partial NS5B gene were performed using BLAST and MEGA version 6. Results: Based on the phylogenetic tree analysis using 360 nucleotides as the partial NS5B gene, Indonesian BVDV-1 isolates from Central and East Java were subdivided to BVDV-1a (n=9), BVDV-1b (n=1), and BVDV-1c (n=5). In the present study, the homology of BVDV subgenotype -1a, -1b, and -1c was compared to the BVDV GenBank data and found 90-93%, 93%, and 92-95% respectively with the average pairwise distance of 0.207. A point mutation was shown at R283K of all BVDV isolates based on the sequence of three amino acid residues R283, R285, and I287 in the nucleotide-binding pocket as a part of the encoded RNA-dependent RNA polymerase. Conclusion: This study revealed the genetic variability of BVDV infecting cattle in Central Java and East Java, Indonesia, the subtypes BVDV-1a, BVDV-1b, BVDV-1c, and a point mutation at the R283K residue.

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The development of Jembrana disease vaccine is importance to prevent the loss of Bali cattle industry in Indonesia. This study aims to prepare a Jembrana DNA vaccine. The data Tat protein sequences gained from NCBI and the consensus process has been finished by the MultAlign program, and then Cloning of the pcDNA3.1-tat has been successfully performed on E. coli DH5α and confirmed by PCR, restriction analysis and sequencing. The propagated plasmids were prepared as DNA-chitosan complex and physiochemical characterized using Particle Size Analyzer. Complex with a 1:2 (wt/wt) ratio of DNA and chitosan have a mean diameter of 268.5 nm and zeta potential +25.1 mV and the value of Cytotoxicity Assay 80-90% as compared to the untreated cells that used as negative control, so it can be concluded that nanoparticles chitosan has good potential as a carrier agent for pcDNA3.1-tat.

Avian influenza virus (AIV) and Newcastle disease virus (NDV) are systemic disease that is highly infectious with high morbidity and mortality rates, and highly detrimental to economic value in the domestic and commercial poultry industries. Generally, in the field there is a mixed infection of AIV and NDV, each of which is highly pathogenic. Clinical signs are difficult to distinguish, so immunopathological immunohistochemistry (IHC) double staining was developed with the ability to confirm the diagnosis of mixed infection. In the present study, 20 chickens infected naturally with clinical signs of torticollis and curled toe paralysis that had a mixture of AIV and NDV infections and there are variations of tropism distribution of AIV and NDV in tissues were used. The AIV and NDV were visualized by performing the double staining technique DAB and Fast-Red with monoclonal antibody anti-hemagglutinin AIV-mouse (AIVHA) and polyclonal antibody antihemagglutinin-neuraminidase NDV-rabbit (NDVHN) proteins. AIV infected-lungs or brains had red coloration (AIVHA) and NDV infected lungs or brains had brown coloration (NDVHN). The IHC double staining technique is sensitive and accurate in the confirmation of the diagnosis of a mixed infection to determine the difference between AIV and NDV.