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All Journal Science: Indonesian Journal of Science Punggawa Global Research
Septianto Wikan Nurhidayat
Universitas Aisyiyah Yogyakarta
Agriculture, Biological Sciences & Forestry Chemistry Mathematics Physics Public Health

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Analisis Filogenetik Cacing Parasit Anisakis spp. Pada ikan di Pesisir Selatan Yogyakarta Septianto Wikan Nurhidayat
Science: Indonesian Journal of Science Vol. 2 No. 2 (2025)
Publisher : Universitas Pahlawan Tuanku Tambusai

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.31004/science.v2i2.351

Abstract

The third-stage larvae of Anisakis spp. are commonly found infecting marine fish, which serve as intermediate hosts. This study aimed to identify Anisakis spp. through sequencing analysis. A total of 15 specimens of hairtail (Trichiurus lepturus) and 15 specimens of red snapper (Lutjanus malabaricus) were examined to detect the presence of Anisakis spp. Fish samples were obtained from fishermen in the Glagah and Trisik, Kulonprogo, Yogyakarta. The examination involved dissection of internal organs, including the body cavity, digestive tract, liver, reproductive tract, and muscle tissue. Larvae of Anisakis were preserved in 70% ethanol. The prevalence of infection in hairtail and red snapper was 33.33% and 6.67%, respectively, with mean intensities of 4.8 and 2 larvae per fish, respectively. Genomic DNA was extracted from four Anisakis larvae using the Wizard Genomic DNA Extraction and Purification Kit (Promega). Polymerase chain reaction (PCR) and sequencing were performed targeting the mitochondrial cox2 gene. Sequence analysis of the cox2 region, combined with phylogenetic analysis, revealed that all examined specimens were identified as Anisakis typica.
Development of aerA Gene-Based PCR Primers as DNA Biomarkers for the Molecular Detection of Aeromonas hydrophila Using an In Silico Approach Septianto Wikan Nurhidayat; Naina Nur Prastiti
Punggawa Global Research: Jurnal Multidisiplin Vol. 1 No. 3 (2026): Punggawa Global Research
Publisher : Punggawa Legacy Center

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.67707/pgr.v1i3.102

Abstract

Aeromonas hydrophila is one of the major bacterial pathogens affecting cultured fish, causing Motile Aeromonas Septicemia (MAS) and substantial economic losses in aquaculture. Early detection of this pathogen is essential for effective disease prevention and control. This study aimed to design a primer pair targeting the aerA virulence gene as a DNA biomarker for the molecular detection of A. hydrophila using an in silico approach. The aerA gene sequence (GenBank accession number EU254217.1) was retrieved from the NCBI database and used as the template for primer design using Primer3Plus. The designed primers were evaluated based on primer length, melting temperature (Tm), GC content, predicted amplicon size, and the potential formation of secondary structures. Primer specificity was subsequently assessed using In Silico PCR against several Aeromonas genomes. The selected primer pair consisted of the forward primer 5'-GAGAAGACGGCCATCAAGGT-3' and the reverse primer 5'-TGTGGTTCCAGTTCGGGC-3', producing a predicted amplicon of 529 bp. The primers exhibited melting temperatures of 59.7°C and 59.9°C with GC contents of 55.0% and 61.1%, respectively, indicating suitable characteristics for PCR amplification. In Silico PCR analysis demonstrated successful amplification exclusively in A. hydrophila genomes, while no amplification was detected in the non-target Aeromonas species evaluated. These findings suggest that the aerA gene is a promising DNA biomarker for the molecular detection of A. hydrophila . However, further in vitro validation is required to confirm the sensitivity and specificity of the designed primers using field isolates.
Co-Authors Naina Nur Prastiti