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.
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.
Sumatran elephant is a subspecies of endemic Asian elephants on the island of Sumatra and is included in the Red List of the International Union for Conservation of Nature (IUCN) with critically endangered status. The building of the Elephant Training Centre (ETC) in Way Kambas National Park (WKNP) is one of the conservation efforts of Sumatran elephants. Small and closed population size lead to an increased risk of inbreeding that triggers reduction in genetic variation and viability and increases the risk of extinction. The phylogenetic pattern of Sumatran elephants in Indonesia has shown a low population genetic diversity. Genetic diversity information is indispensable to support the direction of decision making in Sumatran elephant conservation policy. The DNA isolation of Sumatran elephants in ETC, WKNP has performed as a first step to trace its genetic variation. The advanced step of DNA isolation is the use of cytochrome oxidase subunit I (COI) gene for identification of genetic characteristics in Sumatran elephants. The COI gene is one of the genes on the mitochondrial genome and in molecular studies it is used as a genetic marker to study genetic characteristics between species and individuals. Identification and characterisation are done by sequencing process and data analysis in the form of electroforegram using Molecular Evolution Genetics Analysis (MEGA) software version 6.0. to see the genetic diversity of the female Sumatran elephant population in ETC, WKNP. Based on the results of the analysis it is indicated that the genetic distance of 24 individual female Sumatran elephant from PLG, TNWK is 0.000 with a homology value of 100%, strengthened by the construction of phylogenetic tree. The absence of genetic distance indicates a close genetic relationship, so it can be concluded all individual female Sumatran elephants in the PLG, TNWK is derived from one population group.
Way Kambas National Park (WKNP) is home of five protected big mammals including sumatran elephants. It shares its border with 22 of 37 villages surrounding the national park. Understanding their existence in the wild is a priority, and wildlife genetics is a crucially needed. Besides poaching and habitat fragmentation, wildlife-human conflict is one big issue. Elephant Training Center (ETC) in WKNP is built for semi in-situ conservation effort on captive sumatran elephants that mainly have conflict histories with local people. Participative observation and bio-molecular analysis were conducted to learn the importance of captive Sumatran elephant for conservation effort. Through captive sumatran elephants, database and applicable methods are expected to be developed supporting the conservation of their population in the wild. Participative observation and molecular identification was carried on captive sumatran elephants in ETC, WKNP under multiple year Terapan grant of Ministry of Research and Technology Higher Education, Indonesia. Gene sequence and cytological analyses showed that the captive sumatran elephants are closely related and tend to be domesticated. Translocation among ETC to avoid inbreeding, and maintaining the captive sumatran elephant as natural as possible are highly recommended. Developing genetic database can be a reference for both captive and wild sumatran elephants.
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.
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