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Jembrana disease virus (JDV) is a viral pathogen that causes Jembrana disease in Bali cattle (Bos javanicus) with high mortality rate. An easy and rapid diagnostic method is essential for further control this disease. We used a reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with lateral flow dipstick (LFD), based on conserved tm subunit of Jembrana disease virus env gene. The RT-LAMP conditions were optimized by varying the concentration of MgSO4, betaine, dNTP, and temperature as well as the time and duration of reaction. The primers sensitivity for JDV was confirmed. The method was able to detect env-tm gene dilution which contained 2 × 10(-15) g of template. Comparatively, the sensitivity of RT-LAMP/LFD was 100-fold more sensitive than reverse transcription-polymerase chain reaction. The primers specificity for JDV was also confirmed using positive and negative controls. This work also showed that virus detection could be done not only on total RNA extracted from blood but various organs could also be analyzed for the presence of JDV using RT-LAMP/LFD method. The whole process, including the LAMP reaction and the LFD hybridization step only lasts approximately 75 min. Results of analysis can be easily observed with naked eyes without addition of any chemical or further analysis. The combination of RT-LAMP with LFD makes the method a more suitable diagnostic tool in conditions where sophisticated and expensive equipments are not available for field investigations on Jembrana disease in Bali cattle.

Staphylococcus aureus (S. aureus) is a major cause various infections in humans and animals throughout the world. The increasing incidence of S. aureus infection, particularly methicillin-resistant Staphylococcus aureus (MRSA) is complicated by the antibiotic-resistance phenomenon. Thus, it requires new strategies to prevent wider scale of its incidence occurring. Preventative methods against infectious diseases through vaccination can prevent, control and reduce the severity of the disease process. Various methods have been used in the development of vaccines to prevent S. aureus infections, but no satisfactory results have been obtained. Recent studies suggests that multivalent vaccines based on cell-mediated immunity is expected to provide better protection against infection. The purpose of this article is to summarize the challenges of S. aureus infection and the current status of its potential solution with the development of a multivalent vaccine and to explain the latest approaches recommended to obtain successful development of S. aureus vaccine by inducing T-cells CD8+, CD4+ and Th17. Seven MRSA Indonesian origin isolates from humans and dairy-cattle were used as candidate vaccines as genotypic characterization had previously been performed (coa, eta, etb, hla, hlb, cap5, cap8, fnbA, fnbB and mecA). Vaccination can be performed with live-attenuated or killed-vaccine and could be combined with an injection of αCD40 monoclonal-antibodies (mAbs) with the aim to induce cellular-based active immunity (T CD8+, CD4+ and Th17). These isolates are expected to be safe vaccine candidates with immunogenic-protective properties based on genetic and antigenic variation to prevent and control staphylococcal infections in humans and animals.

Jembrana disease constitutes the main concern in cattle industry especially in Indonesia and Australia as it has caused important economic losses due to mortality of cattle. The pathology of the disease is unusual for a lentivirus infection as it is associated with a severe, often lethal disease syndrome and a short incubation period in cattle. For lack of efficient medical treatment of JDV-infected cattle, vaccination may therefore constitute an effective measure for the prevention or eradication of Jembrana disease. Up to date, only one type of vaccine has been reported and tested. It is based on inactivated, tissue-derived virus antigens (Tabanan/87 isolate, JDVTAB/87). This review summarize show current Jembrana disease vaccine was developed as well as evaluated and how these information might be useful in future vaccine design.

Jembrana disease is a bovine disease that affects Bali cattle (Bos javanicus). The causal agent is named Jembrana Disease Virus (JDV), a lentivirus member. The disease development of Jembrana disease in Bali cattle is unique for a lentivirus infection as it is related with severe clinical syndrome in an acute period. In experimentally JDV-infected Bali cattle, the death rate was about 21% and occurred within only 1 to 2 weeks post-infection. Indeed, the mortality of more than 60 000 cattles in a year was observed during the first outbreak and the disease is now endemic through-out parts of Indonesia. Early diagnosis constitutes a preventive method of further disease outbreaks. The Jembrana disease can be diagnosed by the clinical symptoms but more reliable diagnostic tools are available, based on either antigen (immunodiagnosis) or viral genome (molecular diagnosis). In this review, we summarize about immunodiagnostic tools of Jembrana disease which has been developed so far.

The etiological agent of Jembrana disease is a virus which was accordingly named Jembrana Disease Virus (JDV). JDV was a member of the family of Retroviridae, of the subfamily of lentiviruses. As a member of the lentivirus, JDV has general property to integrate their genome into the chromosomal DNA of the host-cells. It is one of their common properties which is not shared by simple retroviruses such as murine leukimia virus or avian sarcoma virus. Lentiviruses are highly species-specific and share the property of replicating in dividing and non-dividing terminally differentiated cells. JDV-based gene transfer vector which had been developed is appeared to possess a broad tropism and a unique capacity of mediating efficient gene transfer. This review summarize JDV-based viral vector system that has been developed. The main principles and components of JDV viral gene transfer system and its implication in the future research will be discussed.

Jembrana disease virus is a viral pathogen that causes Jembrana disease in Bali cattle (Bos javanicus) with high mortality rate. Infection of Jembrana Disease Virus (JDV) on Bali cattle have caused substantial economic losses for farmers in Indonesia and Australia. In order to control the spread, development of a sensitive detection method is important. In this study, we used three different detection methods based on genomic approach, i.e., reverse transcriptase-polymerase chain reaction (RT-PCR), reverse transcriptase-loop mediated isothermal amplification (RT-LAMP) and dot-blot hybridization to detect JDV. Utilization of pGEX-TM, a recombinant plasmid containing env-tm gene as a positive control showed that RT-LAMP is the most sensitive method compares the two others. It could detect template concentration as low as 10-6 ng µL-1 or equivalent to 1.52A—102 plasmid copy number, 100 and 10000 more sensitive than RT-PCR and dot-blot hybridization, respectively.

1Departement of Reproduction, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan sekip unit II, Yogyakarta, 55281 Indonesia. 2Centre of Biotechnology Study, Gadjah Mada University, Jalan teknika utara, Yogyakarta, 55281 Indonesia. 3Faculty of Medicine, Sebelas Maret University, Jalan Ir. Sutami 36 A, Surakarta, 57126 Indonesia. 4Departement of Microbiology, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan sekip unit II, Yogyakarta, 55281 Indonesia. 5Department of Internal Medicine, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan sekip unit II, Yogyakarta, 55281 Indonesia.

Toxoplasmosis, arising from infection by Toxoplasma gondii, is one of the most common parasitic diseases in humans and other warm-blooded animals. In humans, infections are usually asymptomatic but severe disease can occur in immunocompromized individuals and newborns. Due to the importance of the disease and in order to take suitable measures, an early diagnosis of the disease is essential, particularly for pregnant women and in industry of domestic animals. The genome of T. gondii contains repeat sequences B1 and R522 which constitute ideal targets for genome-based detection methods. The 522 base pairs repeat sequences R522 are the most promising due to the high copy number, evaluated to be 200 to 300 units within the genome. We developed a simple dot-blot hybridization based on R522 sequences. The method is simple and does not require sophisticated devices. The test of the method, using cloned R522 as target, showed that the parasite detection method was sensitive and proved to be promising for use in routine health controls as well as for the survey of Toxoplasma infections. Key words: DIG-probe, dot blot hybridization, repeat sequences, R522, Toxoplasma gondii