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PRIMER OPTIMIZATION AS GENETIC MARKER PRIMER BASED ON CYTOCHROME B GENE ON SUMATRAN ELEPHANT (Elephas maximus sumatranus) NON-INVASIVE SAMPLES Eko Agus Srihanto; Elly Lestari Rustiati; Priyambodo Priyambodo; Dian Neli Pratiwi; Alvin Wiwiet Susanto; Diah Esti Anggraini; Enny Saswiyanti
BIOVALENTIA: Biological Research Journal Vol. 8 No. 1 (2022)
Publisher : Biology Department, Faculty of Mathematics and Natural Sciences, Sriwijaya University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (773.02 KB) | DOI: 10.24233/biov.8.1.2022.216

Abstract

Sumatran elephant is declared as critically endangered by IUCN since 2011. In supporting conservation efforts on sumatran elephant in its natural habitat, molecular data collection of their population needs to be done. Genetic source material is mostly obtained in its natural habitat is its dung/feces. Study on cytochrome B gene as genetic marker from sumatran elephant fecal samples can be done to get additional genetic data as a form of conservation effort. Primer optimization as genetic marker based on cytochrome B gene has been carried out as an early study on sumatran elephant genetics.  Primer optimization program based on cytochrome B genetic marker from sumatran elephant fecal samples was under Higher Education Applied Research funded by The Ministry of Research, Technology, and Higher Education. DNA was isolated from fecal samples of captive sumatran elephants in Elephant Training Center, Way Kambas National Park. Primer optimization was done via amplification by increasing certain temperature. Specificity test was done to ensure that the primer only recognized sumatran elephant DNA. The best result was obtained with the annealing temperature of 52oC
Species Confirmation for Robusta Coffee in Sedayu, Semaka, Tanggamus: Social Forestry Data Base Management Elly Lestari Rustiati; Priyambodo Priyambodo; Eko Agus Srihanto; Dian Neli Pratiwi; Alvin Wiwiet Susanto; Muhammad Febriansyah; Andriyani Wijaya Kusuma; Enny Saswiyanti
Jurnal Ilmu Lingkungan Vol 23, No 4 (2025): July 2025
Publisher : School of Postgraduate Studies, Diponegoro Univer

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jil.23.4.1149-1154

Abstract

Lampung, including the Register 31 Pematang Arahan area which is next to the Bukit Barisan Selatan National Park, is known for its high biodiversity, and its coffee production. Gapoktanhut Lestari Sejahtera, consisting of 13 Forest Farmer Groups (KTH), practiced community-based forest coffee under the KPH Kotaagung Utara, Tanggamus, Lampung. To support the development of biodiversity data base for KPH Kotaagung Utara, molecular-based species identification for robusta coffee has been carried out under BLU LPPM Unila grant year 2023 and in collaboration with the Lampung Disease Investigation Center. Robusta coffee leaf samples from six KTH areas (Bumi Mulyo, Sido Makmur 1, Sido Makmur 2, Sido Makmur 3, Mandiri Jaya, and Murah Rejeki) were collected for molecular species confirmation based on the N-methyl transferase genetic marker. Molecular analysis included DNA extraction, amplification, and electrophoresis conducted at the Lampung Disease Investigation Center. Sequencing was based on the Coffea canephora CAF2 genetic marker. Bioinformatics analysis was performed using the Basic Local Alignment Search Tool (BLAST) and Molecular Evolutionary Genetics Analysis (MEGA) software version 6. Identical sequence lengths of samples in 6 KTH are varied in base pair length and nitrogen base sequences, ranging from 163-530 bp, with 5 out of 6 KTH having a genetic distance of 0.000%, while 1 sample had a genetic distance of 0.006%. It indicates that robusta coffee samples from 6 KTH areas are genetically closely related. It is supported by its phylogenetic tree, the coffee species cultivated by Gapoktanhut Lestari Sejahtera is molecularly confirmed to be robusta coffee, Coffea canephora.
Learning RNA Isolation on Oral Swab Sample on Bats in Bandar Lampung Urban Areas Elfita Nova Yunior; Elly Lestari Rustiati; Enny Saswiyanti; Emantis Rosa; Priyambodo Priyambodo; Eko Agus Srihanto; Dian Neli Pratiwi
Jurnal Agrosci Vol 3 No 4 (2026): Vol 3 No 4 March 2026
Publisher : Ann Publisher

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.62885/agrosci.v3i4.1166

Abstract

Background. Bats are recognized as important reservoir hosts for various zoonotic viruses, including coronaviruses, posing potential public health risks, particularly in urban environments where human–animal interactions are frequent. Aims. This study aimed to evaluate the effectiveness of RNA isolation from oral swab samples of bats captured in urban areas of Bandar Lampung, Indonesia, as a preliminary step for molecular detection of viral pathogens. Methods. Sampling was conducted using mist nets in the University of Lampung area, followed by oral swab collection and preservation in viral transport medium. RNA isolation was performed through four main stages: lysis, binding, washing, and elution. The quality of RNA was assessed using agarose gel electrophoresis, while the quantity was measured using a Qubit Fluorometer. Result. A total of six bats were successfully sampled, consisting of Cynopterus brachyotis and Cynopterus sphinx. Qualitative analysis revealed a single thin, luminescent band in all samples, indicating suboptimal RNA integrity. Quantitative results showed RNA concentrations ranging from 0.58 to 2.03 ng/µL, with three samples falling within the acceptable range (1.8–2.0 ng/µL), suggesting adequate RNA purity. Variations in RNA quality and concentration were likely influenced by factors such as incubation duration and sample handling, potentially leading to contamination or degradation. Conclusion. In conclusion, RNA isolation from bat oral swabs in urban areas is feasible; however, protocol optimization is necessary to improve RNA integrity and yield. High-quality RNA is essential for reliable downstream molecular analyses, including viral detection and surveillance of zoonotic diseases
Oral Swab-Based DNA Extraction Of Cynopterus Brachyotis: An Initial Step For Species Confirmation Nabila Aulia Rhamadaningtyas; Elly Lestari Rustiati; Priyambodo Priyambodo; Eko Agus Srihanto; Enny Saswiyanti; Dian Neli Pratiwi; Alvin Wiwiet Susanto
Jurnal Agrosci Vol 3 No 3 (2026): Vol 3 No 3 January 2026
Publisher : Ann Publisher

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.62885/agrosci.v3i3.1025

Abstract

Background. Identifying bat species based on morphological characteristics often faces challenges due to character overlap among species, especially within cryptic species groups. Cynopterus brachyotis is one of the fruit bats with high genetic diversity but relatively uniform morphology. Aims. Therefore, a molecular approach is needed to support accurate species confirmation. Under the HETI Research Grant of Innovation and Collaboration Batch 3- II Year 2025, this study aims to obtain genomic DNA from C. brachyotis bats caught at the Lampung Disease Investigation Center as the initial step of molecular species confirmation. Methods. Samples were obtained by taking oral swabs on bat individuals caught using mist nets. DNA extraction is performed using silica membrane-based methods with commercial kits. Quantitative evaluation of DNA was performed using a Qubit Fluorometer, while qualitative evaluation was performed by agarose gel electrophoresis. Conclusion. The results showed that the DNA concentration was in the low range (0.1–0.7 ng/μL), with a DNA band appearing very thin on electrophoresis. Implementation. Nevertheless, the quantification results confirm that DNA was successfully extracted and remains suitable for advanced molecular analysis, with optimization at the DNA amplification stage.
DNA Extraction of Cynopterus brachyotis of Labuhan Ratu VII Based on Column Method in Supporting its Molecular Species Confirmation Annisa Lidya Maharani; Elly Lestari Rustiati; Priyambodo Priyambodo; Eko Agus Srihanto; Enny Saswiyanti; Dian Neli Pratiwi; Alvin Wiwiet Susanto
Jurnal Agrosci Vol 3 No 1 (2025): Vol 3 No 1 September 2025
Publisher : Ann Publisher

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.62885/agrosci.v3i1.1043

Abstract

Background. Bats (Chiroptera) are true flying mammals and nocturnal. Ecologically, bats play important roles as pollinators, seed dispersers, guano producers, biocontrol, and hosts a range of viruses. Cynopterus brachyotis belongs to fruit-eating bats and is considered morphologically cryptic with C. sphinx. Aims. To support morphological identification, molecular species confirmation needed to be conducted. DNA extraction, as an initial step, plays a crucial role in molecular analysis. Methods. The quality and quantity of extracted DNA determine the success rate of sequencing for species confirmation in bats. In this study, oropharyngeal samples were used. Result. The research procedures included bat capture and DNA oropharyngeal swab sample collection, sample preparation, DNA extraction using a silica column-based commercial kit, DNA concentration measurement using a Qubit assay, and electrophoresis. Conclusion. The Qubit assay showed DNA concentrations of >1 ng/µL, while electrophoresis did not reveal bright DNA bands.
Learning Bats Handling for Oral Swab Sampling Sevira Nur Azmi; Elly Lestari Rustiati; Priyambodo Priyambodo; Enny Saswiyanti; Eko Agus Srihanto; Dian Neli Pratiwi; Jani Master; Waryoko Susandi
Jurnal Agrosci Vol 3 No 4 (2026): Vol 3 No 4 March 2026
Publisher : Ann Publisher

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.62885/agrosci.v3i4.1154

Abstract

Background. The only flying mammals, bats play an essential role in their natural habitat. Based on the type of food, bats can be divided into two groups: fruit-eating bats and insect-eating bats. Bats rank second-highest in species diversity, with 1,439 worldwide and 239 in Indonesia. Known as reservoirs of viruses, including coronaviruses. Bats are suspected to be related to COVID-19. Aims. Under the Research Innovation and Collaboration Program - Higher Education for Technology and Innovation Project (HETI) University of Lampung 2024-2025, and in collaboration with the Lampung Disease Investigation Centre, learning the procedures for handling bats properly so as not to physically harm bats in oral swab sampling and species recognition in Braja Harjosari, directly next to Way Kambas National Park was done. The life-trapping technique uses a mist net. Methods. Bat handling for taking an oral swab was carried out using the pinch grip method, holding both bat arms backwards with the thumb and middle finger, with the bat positioned facing upwards. Conclusion. The bat's mouth is then blown open, and a cotton swab is gently inserted into the bat's mouth. Afterwards, the bat is rested and given a drink/water and released into nature. Oral swab samples from 10 individual bats, fruit-eating bats, Cynopterus brachyotis (n = 8), Cynopterus horsfieldii (n = 1), and an insect-eating bat, Scotophilus kuhlii (n = 1).
RNA Isolation from Oral Swab Sample On Bats of Rural Area, Braja Harjosari, East Lampung Vidyanti Kurniasih; Elly Lestari Rustiati; Priyambodo Priyambodo; Eko Agus Srihanto; Enny Saswiyanti; Dian Neli Pratiwi
Jurnal Agrosci Vol 3 No 3 (2026): Vol 3 No 3 January 2026
Publisher : Ann Publisher

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.62885/agrosci.v3i3.1161

Abstract

Background. Bats are considered animals that carry infectious diseases, are known to be reservoirs of viruses, and are suspected of causing disease outbreaks. The World Health Organization stated that this disease outbreak is caused by Coronavirus Disease 2019 (COVID-19), which is caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). Aims. Under the Research Innovation and Collaboration Program - Higher Education for Technology and Innovation Project (HETI) and in collaboration with the Lampung Disease Investigation Center, a preliminary study on EID detection on bats based on Predict Protocols was done to identify the presence of coronavirus on bats, including rural areas, Braja Harjosari, East Lampung. It is located next to Way Kambas National Park. Methods. Oral swab samples were taken from ten bats, 3 species: Cynopterus brachyotis and Cynopterus horsfieldi, and Scotophilus kuhlii. RNA extraction was conducted by Predict Protocols, which has four stages: lysis, binding, washing, and elution. Conclusion. Two samples showed thin-band luminescence. While in the quantity test, there are three samples of good concentration results between 1.8 and 2.0.