Ekowati Handharyani

Molecular detection of a novel paramyxovirus in fruit bats from Indonesia

BackgroundFruit bats are known to harbor zoonotic paramyxoviruses including Nipah, Hendra, and Menangle viruses. The aim of this study was to detect the presence of paramyxovirus RNA in fruit bats from Indonesia.MethodsRNA samples were obtained from the spleens of 110 fruit bats collected from four locations in Indonesia. All samples were screened by semi-nested broad spectrum reverse transcription PCR targeting the paramyxovirus polymerase (L) genes.ResultsSemi-nested reverse transcription PCR detected five previously unidentified paramyxoviruses from six fruit bats. Phylogenetic analysis showed that these virus sequences were related to henipavirus or rubulavirus.ConclusionsThis study indicates the presence of novel paramyxoviruses among fruit bat populations in Indonesia.

Canine Hemangiopericytoma: An Evaluation of Metastatic Potential

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Isolation and characterization of a novel alphaherpesvirus in fruit bats.

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Detection of novel polyomaviruses in fruit bats in Indonesia

ABSTRACT Bats are known to harbor emerging RNA viruses. Recent studies have used high-throughput sequencing technology to identify various virus species, including DNA viruses that are harbored by bats; however, little is known about the nature of these potentially novel viruses. Here, we report the characterization of a novel herpesvirus isolated from an Indonesian pteropodid bat. The virus, tentatively named fruit bat alphaherpesvirus 1 (FBAHV1), has a double-stranded DNA genome of 149,459 bp. The phylogenetic analyses suggested that FBAHV1 is phylogenetically grouped with simplexviruses within the subfamily Alphaherpesvirinae. Inoculation of FBAHV1 into laboratory mice caused a lethal infection. Virus infection was observed in lung, liver, and brain tissue. Serological and PCR screening revealed that fruit bats infected with FBAHV1 or its related virus are widely distributed in Indonesia. The identification of FBAHV1 makes a considerable contribution to our understanding of simplexviruses associated with bats. IMPORTANCE Bats are known to harbor emerging viruses, such as lyssaviruses, henipaviruses, severe acute respiratory syndrome-like coronaviruses, and filoviruses. Although alphaherpesviruses are disseminated in humans and other animals, there is little information about their distribution in bats. Here, we isolated a previously unknown alphaherpesvirus from an Indonesian fruit bat. Genome sequence analysis suggested that the virus is a member of the genus Simplexvirus within the subfamily Alphaherpesvirinae, which also includes common human viruses, such as herpes simplex virus 1 and herpes simplex virus 2. FBAHV1 is the first bat-derived alphaherpesvirus whose complete genome has been sequenced.

Epidemiological study of fowl glioma-inducing virus in chickens in Asia and Germany

Bats are an important natural reservoir for a variety of viral pathogens, including polyomaviruses (PyVs). The aims of this study were: (i) to determine which PyVs are present in bats in Indonesia and (ii) to analyze the evolutionary relationships between bat PyVs and other known PyVs. Using broad-spectrum polymerase chain reaction (PCR)-based assays, we screened PyV DNA isolated from spleen samples from 82 wild fruit bats captured in Indonesia. Fragments of the PyV genome were detected in 10 of the 82 spleen samples screened, and eight full-length viral genome sequences were obtained using an inverse PCR method. A phylogenetic analysis of eight whole viral genome sequences showed that BatPyVs form two distinct genetic clusters within the proposed genus Orthopolyomavirus that are genetically different from previously described BatPyVs. Interestingly, one group of BatPyVs is genetically related to the primate PyVs, including human PyV9 and trichodysplasia spinulosa-associated PyV. This study has identified the presence of novel PyVs in fruit bats in Indonesia and provides genetic information about these BatPyVs.

Divergent bufavirus harboured in megabats represents a new lineage of parvoviruses.

Fowl glioma-inducing virus (FGV), which belongs to avian leukosis virus (ALV) subgroup A, induces fowl glioma. This disease is characterized by multiple nodular gliomatous growths of astrocytes and has been previously reported in Europe, South Africa, Australia, the United States and Japan. FGV and FGV variants have spread to ornamental Japanese fowl, including Japanese bantams (Gallus gallus domesticus), in Japan. However, it is unclear how and where FGV emerged and whether FGV is related to the past fowl glioma in European countries. In this study, the prevalence of FGV in European, Asian and Japanese native chickens was examined. FGV could not be isolated from any chickens in Germany and Asian countries other than Japan. Eighty (26%) out of 307 chickens reared in Japan were positive by FGV-screening nested polymerase chain reaction and 11 FGV variants with an FGV-specific sequence in their 3′ untranslated region were isolated. In addition, four other ALVs lacking the FGV-specific sequence were isolated from Japanese bantams with fowl glioma and/or cerebellar hypoplasia. These isolates were considered to be distinct recombinant viruses between FGV variants and endogenous/exogenous avian retroviruses. These results suggest that the variants as well as distinct recombinant ALVs are prevalent among Japanese native chickens in Japan and that FGV may have emerged by recombination among avian retroviruses in the chickens of this country.

Pathotypic characterization of Newcastle disease virus isolated from vaccinated chicken in West Java, Indonesia

Bufavirus is a recently recognized member of the genus Protoparvovirus in the subfamily Parvovirinae. It has been reported that human bufavirus was detected predominantly in patients with diarrhoea in several countries. However, little is known about bufavirus or its close relatives in nonhuman mammals. In this study, we performed nested-PCR screening and identified bufavirus from 12 megabats of Pteropus spp. in Indonesia. Furthermore, we determined nearly the full genome sequence of a novel megabat-borne bufavirus, tentatively named megabat bufavirus 1. Phylogenetic analyses showed that megabat bufavirus 1 clustered with known protoparvoviruses, including human bufavirus but represented a distinct lineage of bufavirus. Our analyses also inferred phylogenetic relationships among animal-borne bufaviruses recently reported by other studies. Recombination analyses suggested that the most common recent ancestor of megabat bufavirus 1 might have arisen from multiple genetic recombination events. These results characterized megabat bufavirus 1 as the first protoparvovirus discovered from megabats and indicates the high genetic divergence of bufavirus.

Pathology and immunohistochemistry study of Newcastle disease field case in chicken in Indonesia

Aim: This research was conducted to differentiate and characterize eight Newcastle disease virus (NDV) isolates collected from vaccinated chicken at commercial flocks in West Java, Indonesia, in 2011, 2014 and 2015 by pathotype specific primers. Materials and Methods: A total of eight NDV isolates collected from clinical outbreaks among commercial vaccinated flocks in West Java, Indonesia, in 2011, 2014, and 2015 were used in this study. Reverse transcription-polymerase chain reaction was used to detect and differentiate virulence of NDV strains, using three sets of primers targeting their M and F gene. First primers were universal primers to detect NDV targeting matrix (M) gene. Other two sets of primers were specific for the fusion (F) gene cleavage site sequence of virulent and avirulent NDV strains. Results: Our results showed that three isolates belong to NDV virulent strains, and other five isolates belong to NDV avirulent strains. The nucleotide sequence of the F protein cleavage site showed 112K/R-R-Q/R-K-R/G-F117 on NDV virulent strains and 112G-K/R-Q-G-R-L117 on NDV avirulent strain. Conclusion: Result from the current study suggested that NDV virulent strain were circulating among vaccinated chickens in West Java, Indonesia; this might possess a risk of causing ND outbreaks and causing economic losses within the poultry industry.

Subjective and objective observation of skin graft recovery on Indonesian local cat with different periods of transplantation time

Aim: The aim of the study was to examine pathology and the distribution pattern of Newcastle disease virus (NDV) in internal organs of chickens from a field case using immunohistochemical staining. Materials and Methods: 10 groups of broiler, layer, and domestic chicken were collected from necropsy room Division of Pathology, Bogor Agricultural University. These chickens were originated from West Java and collected based on pathologist diagnosis as suspect of Newcastle disease (ND). They were subsequently confirmed positive of ND with real-time-reverse transcription polymerase chain reaction assay. The respiratory, circulatory, digestive, lymphoreticular and central nervous systems were collected for histopathology examination. Results: The gross pathology and histopathology changes were tracheitis, pneumonia, pericarditis, myocarditis, catarrhal proventriculitis, catarrhal enteritis, typhlitis, perihepatitis, pancreatitis, nephritis interstitial, splenitis, atrophy of Bursa Fabricius, and encephalitis. Conclusion: The distribution pattern of NDV in internal organs of chickens from a field case in this study is similar with a previous reported pattern in systemic cases of the internal chicken organs. High intensity of immunohistochemistry stain result was detected in trachea, lung, proventriculus, duodenum, cecal tonsil, kidney, and brain.

Expression of SC1, a cell adhesion molecule, promotes the metastatic activities of the Gallus gallus lymphoblastoid cell line MDCC-MSB1 derived from Marek's disease

Aim: The success of a skin graft in a cat is highly dependent on the granulation formed by the base of recipient bed. Granulation by the base of recipient bed will form after several days after injury. This research aimed to observe subjective and objective profile of skin graft recovery on forelimb of cats with different periods of donor skin placement. Materials and Methods: Nine male Indonesian local cats aged 1-2 years old, weighing 3-4 kg were divided into three groups. The first surgery for creating defect wound of 2 cm×2 cm in size was performed in the whole group. The wound was left for several days with the respective interval for each group, respectively: Group I (for 2 days), Group II (for 4 days), and Group III (for 6 days). In the whole group, the second surgery was done by the harvesting skin of thoracic area which then applied on recipient bed of respective groups. Result: The donor skin on Group II was accepted faster compared to Group I and Group III. The donor skin did not show color differences compared to surrounding skin, painless, bright red in bleeding test had faster both hair growth and drug absorption. Test toward the size of donor skin and the effect of drugs did not show a significant difference between each group. Conclusion: The observe subjective and objective profile of skin graft recovery on forelimb of cats on Group II were accepted faster compared to Group I and III.