Molecular Characterization of Anguilla from Cibereum and Sapuregel Rivers Segara Anakan Watersheds Cilacap, Central Java

Abstract

The taxonomic status of Anguilla species' in river watershed that empties into Segara Anakan Cilacap is uncertain, thereby making it difficult for further studies to be carried out to determine its genetic in that area. Therefore, this study evaluates Anguilla's taxonomic status and population genetic in Cibeureum and Sapuregel River watersheds. Data were obtained from molecular characterization study using cytochrome c oxidase 1, with fourteen Anguilla specimens collected from two sequenced watersheds. Taxonomic status was determined based on homology and divergence values and monophyly of the samples to the reference species. Meanwhile, genetic divergences among samples to the reference species were calculated based on the Juke-Cantor substitution model in DnaSP6. A homology test was performed using a basic local alignment search tool, with monophyly inferred from the cladogram, which was developed using neighbor-joining and maximum likelihood algorithms in MEGAX with 1000 pseudoreplicates and out-group comparison. Furthermore, population genetic was analyzed through polymorphism, haplotype, nucleotide diversity within the population, divergence, and genetic differences. All calculations conducted in Arlequin 3.5 had Anguilla samples comprising of high (99.23% to 99.84%) to low genetic divergences (0.224% to 1.127%). The result shows that cladogram with all Anguilla samples formed a monophyletic clade with A. bicolor, separated from their taxa. Furthermore, Anguilla samples from both watersheds have low genetic polymorphisms with medium to high haplotype and nucleotide diversity. The population's comparison proved that both populations have low genetic divergence, and no genetic difference based on variance analysis (p=0761). Therefore, Anguilla resources in river watersheds that empty into Segara Anakan are a single genetic conservation unit.

Morphological and molecular identification of java barb (Barbonymus gonionotus) from East Java Province

“…Furthermore, preservation is carried out with 90% alcohol and stored in a -20C freezer for further analysis [11]. DNA isolation was carried out using the gSYNC TM DNA Extraction kit [12]. The quality of DNA isolation was observed on 1.2% agarose gel using electrophoresis, then stored in a -20 C freezer.…”

Application of the Cytochrome-b gene region in Population Study of Striped Snakehead (Channa striata) by Molecular Approach

“…DNA extraction using the gSYNCTM DNA Extraction kit (Maulid et al 2016, Nuryanto et al 2020. Amplification and visualization of DNA fragments was carried out by Polymerase Chain Reaction (PCR) using 2 pairs of universal primers.…”

“…The second primer, used for species identification using DNA barcoding regions in the COI gene using a pair of Primers FishF1: 5' TCA ACC AAC CAC AAA GAC ATT OGC AC 3') and FishR1: 5' TAG ACT TCT GGG TGG CCA AAG AAT CA 3' (Panprommin et al 2019). The PCR conditions include the predenaturation stage of 94 o C for 2 minutes, the denaturation process at 94 o C for 15 seconds, the annealing or attachment process at a temperature of 50 o C for 15 seconds and the extension process at 72 o C for 30 seconds and the post-extension process for 2 minutes at a temperature of 72 o C. The PCR product is then sent to the PT Genetics Science Indonesia company for the sequencing stage process (Nuryanto et al 2020).…”

Capture-based aquaculture of glass eel nurseries, Anguilla bicolor in Segara Anakan, Cilacap, Central Java, Indonesia

“…Glass eel is the "seed stock" of choice for Asian and European commercial eel cultivation, which relies on dwindling natural stocks. The International Union for Conservation of Nature (IUCN) noted that the global status of Anguilla bicolor and A. marmorata is NT (Near Threatened) and LC (Least Concern), respectively the change in the conservation status of eels from Least Concern to Near Threatened in 2010, 2017, and 2020 is of concern to all of us [19,20].…”