DNA extraction procedures may exhibit different levels of sensitivity. A preliminary effort to obtain DNA from captive Sumatran elephants (Elephas maximus sumatranus) in Elephant Training Center Way Kambas National Park, Lampung for building their molecular individual identification was done. DNA of 30 male and 23 female captive elephants’ blood samples was extracted by QIAGEN. Qualitative analysis of DNA extraction of captive elephant was conducted using two methods, simple electrophoresis and electrophoresis based on PCR product. The aim of this project is to compare those two methods of qualitative analysis of DNA extraction. The first method used 1% agarose gel electrophoresis in the TAE buffer (Tris-Acetate-EDTA), while the second method used Polymerase Chain Reaction (PCR) technique with Glyseraldehyde-3-Phosphate Dehydrogenase (GAPDH) primer. The simple electrophoresis showed 41.5 % positive samples, while the second method showed 86.7% positive samples. The electrophoresis based on PCR product exhibited more sensitivity to detect the DNA from blood of each captive elephant.
Background and Aim: Newcastle disease (ND) is a viral infectious disease that affects commercial and native chickens, resulting in economic losses to the poultry industry. This study aimed to examine the viral strains circulating in commercial and native chickens by genetic characterization and observe the distribution of Newcastle disease virus (NDV) in chicken embryonic tissue. Materials and Methods: ND was detected using a quantitative reverse transcription-polymerase chain reaction. Genetic characterization of the fusion (F) and hemagglutinin-neuraminidase (HN) genes from the eight NDVs was performed using specific primers. The sequence was compared with that of other NDVs from GenBank and analyzed using the MEGA-X software. The distribution of NDV in chicken embryos was analyzed based on lesions and the immunopositivity in immunohistochemistry staining. Results: Based on F gene characterization, velogenic NDV strains circulating in commercial and native chickens that showed varying clinical symptoms belonged to genotype VII.2. Lentogenic strains found in chickens without clinical symptoms were grouped into genotype II (unvaccinated native chickens) and genotype I (vaccinated commercial chickens). Amino acid variations in the HN gene, namely, the neutralization epitope and antigenic sites at positions 263 and 494, respectively, occurred in lentogenic strains. The NDV reaches the digestive and respiratory organs, but in lentogenic NDV does not cause significant damage, and hence embryo death does not occur. Conclusion: This study showed that velogenic and lentogenic NDV strains circulated in both commercial and native chickens with varying genotypes. The virus was distributed in almost all organs, especially digestive and respiratory. Organ damage in lentogenic infection is not as severe as in velogenic NDV. Further research is needed to observe the distribution of NDV with varying pathogenicity in chickens.
Newcastle disease (ND) is a contagious disease in poultry and numerous birds of various ages. Eagle is considered a potential reservoir for ND transmission as a wild bird. This research was conducted to molecularly characterize Newcastle disease virus (NDV) isolated from ND cases in Brontok Eagle and analyze the pathogenesis in chicken embryos. qRT-PCR was conducted as confirmation of NDV without mixing Avian Influenza (AI). Sequencing the fusion (F) and haemagglutinin-neuraminidase (HN) genes from the three NDVs was performed with a specific primer. Amino acid sequence compared with other NDV from Genbank. Pathogenicity, genetic variation, distance, and phylogenetic studies were analyzed using bioinformatics software (MEGA-X). This study analyzed pathogenesis based on lesions and distribution of viral antigens in chicken embryos infected with NDV. Observations were based on tissue lesions with HE and IHC staining. NDV isolated from three Brontok Eagles is classified as velogenic strain, virulent NDV (KRQKRF), and belonging to Genotype VII subgenotype VII.2. The NDV was detected in various organ lesions, more severe in the pulmo, trachea, proventriculus, and intestine of chicken embryos. That is still similar to the previous case reports in the field. These results show that NDV, which infected Brontok Eagle, has similar molecular characteristics and pathogenesis in chickens. These cases could be a threat to the poultry industry. Further research, surveillance, and monitoring of wild birds are needed to obtain more NDV epidemiological information in wild birds.
Elephant Training Center (ETC) Way Kambas National Park (WKNP) was built to support human-elephantmitigation conflict. The small population of captive sumatran elephant in ETC WKNP need a comprehensivestrategy in order to maintain the genetic variation of each individual and avoid inbreeding drive. Currently, geneticstudies have opened new field studies in ecology, included conservation ecology. Patterns in variation of populationhas been investigated by molecular method supporting species conservation effort. The captive sumatran elephant’sID Card is a necessary in database building, which included morphology, health status, and genetic profile. Geneticprofile in each ID Card was filled by cytogenetic and molecular profile for RADP result, that initiated with DNAisolation. The DNA sources collected by blood sampling protocol described by Asiyah et al. (2016) from captivesumatran elephant in ETC, WKNP, and be carried to laboratory in cold condition. The DNA sources stored at 4oCand isolated following commercial protocol. The result of DNA isolation stored at -20oC until amplificationanalysis. DNA isolation was successfully done, for further individual genetic ID building.
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