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"ABSTRAKPemantauan spesies asing invasif berperan penting dalam mempelajari konservasi dan pengendalian ekosistem perairan. Alligator gar merupakan salah satu spesies asing yang masuk ke dalam perairan Indonesia dan berpotensi invasif. Sifatnya yang kriptik menyebabkan spesies ini sulit diamati secara visual. Metode eDNA hadir sebagai alternatif untuk deteksi spesies. Namun penelitian eDNA umumnya dilakukan di daerah beriklim subtropis. Pengembangan metode eDNA di daerah tropis dilakukan dengan menggunakan sedimen kolam artifisial untuk deteksi alligator gar. Sedimen diharapkan mampu melindungi eDNA dari degradasi akibat faktor lingkungan. Pengambilan sampel dilakukan empat kali dalam sebulan dengan mengeruk langsung lapisan sedimen sebanyak 500 mg dan diisolasi menggunakan Fast DNA Spin Kit for Soil. Hasil spektrofotometri menunjukkan konsentrasi terbesar yaitu 204,37 ng/ L pada sampel 1. Suhuannealing telah dioptimasi yaitu 53.4 C menggunakan ecoPrimer dengan gen target 12SrRNA, pita paling jelas ditunjukkan oleh sampel 4 dengan ukuran amplikon 115 bp. Hasil blastn menunjukkan kekerabatan tertinggi pada dua spesies yaitu A. tropicus dan A.spatula, mengindikasikan bahwa sampel sedi mendapat digunakan untuk deteksi alligator gar A. spatula.

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ABSTRACTMonitoring of alien invasive species plays an important role in management and preservation of natural ecosystem. Alligator gar is an alien species in Indonesia swater and potentially invasive. Its cryptic and solitary nature makes visual detection difficult, there fore can not be monitored using traditional method. Recent advances studies have produced technology called eDNA, which allows species detection by using environmental samples such as water, sediment or soil. However, this study often carried out in subtropical area. Development of eDNAin tropical aquatic was performed by using sediment to detect alligator gar. Insediment, the persistance of eDNA can be much longer due to slow rate of degradation. Sample collection was done four times in artificial pond by dredging 500 mg of sediment for each sample. Samples were processed directly using FastDNA Spin Kit for soil. Electrophoresis and spectrophotometry results show edall samples positive containing eDNA, the largest concentration 204,37 ng Lbelong to sample 1. Annealing temperature was optimized at 53.4 C by using ecoPrimer with target region 12 SrRNA, the clearest band shown by sample 4, with the size 115 bp. Blastn result showed the highest similarity with two species A. tropicus and A.spatula. The result was indicating that eDNA contained insediment samples allows for detection of alligator gar A. spatula."

"ABSTRAKIntroduksi spesies asing merupakan ancaman bagi banyak organisme perairan. Deteksi dini dapat meningkatkan keberhasilan usaha pengendalian spesies asing, namun saat ini belum ada metode konvensional yang mampu melakukan hal tersebut. Pendekatan terbaru menggunakan environmental DNA eDNA mulai digunakan sebagai pendukung metode survey konvensional, terutama di daerah beriklim sedang temperate. Suhu yang lebih rendah diperkirakan dapat mempertahankan keberadaan DNA di dalam air. Penelitian ini dilakukan untuk menguji metode eDNA di daerah beriklim tropis untuk mendeteksi spesies asing alligator gar Atractosteus spatula dilakukan di Depok, Indonesia. Sebanyak 200 ml sampel air diambil dari kolam artifisial setiap tujuh hari selama satu bulan, lalu difiltrasi melalui kertas filter nitrat selulosa berdiameter 0,45. Sampel diekstraksi berdasarkan protokol kit FastDNA Spin Kit for Soil. Sampel kemudian diamplifikasi dengan primer ecoPrimer yang memiliki gen target 12S rRNA, lalu dikuantifikasi dan dilakukan sekuensing. Hasil penelitian menunjukkan bahwa eDNA terdeteksi di tiga dari empat sampel air dan mengindikasikan bahwa metode eDNA dapat dipertimbangkan sebagai metode pendukung bagi metode survey konvensional. Meskipun demikian, penelitian lebih lanjut diperlukan sebelum mengaplikasikan metode tersebut di lapangan.

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ABSTRACTIntroduction of non native species threatens the life of many aquatic organisms. Successful eradication requires early detection, but currently no traditional monitoring technique offer the quality to do so. New approach using environmental DNA eDNA begins to be used to complement the more traditional monitoring methods. Most of eDNA studies were carried out in temperate areas, as the cooler temperature preserve the DNA better. This study aims to test the applicability of eDNA method in detecting alligator gar Atractosteus spatula from an artificial pond in Depok, Indonesia. 200 mL water samples was taken every seven days in a month before being filtered through 0,45 nitrate cellulose filter. DNA was isolated using FastDNA Spin Kit for Soil for the subsequent PCR process. Samples were then amplified using ecoPrimer which targeted 12S rRNA gene and quantified using spectrophotometer and electrophoresis, and sequenced. The study showed that eDNA was detected in three out of four samples, and therefore should be considered to complement the more traditional monitoring methods. However, further study is still needed before this method can be widely applied in the field."

"Kura-kura brazil (Trachemys scripta elegans) merupakan salah satu spesies invasif yang berasal dari Amerika Selatan dan telah berkembang di berbagai negara akibat dari pelepasan hewan peliharaan serta dapat berdampak buruk bagi spesies asli. Penelitian dilakukan untuk mendeteksi keberadaan spesies Trachemys scripta elegans pada enam danau pada wilayah Universitas Indonesia, Depok, Jawa Barat. Pendeteksian spesies tersebut dilakukan dengan metode environmental DNA (eDNA) dan quantitative PCR (qPCR) dengan gen target 12S rRNA. Materi genetik telah diisolasi menggunakan metode PCI dan gen target 12S rRNA dikuantifikasi menggunakan qPCR. Keberadaan gen target pada sampel ditentukan melalui nilai LoD dan LoQ yang didapatkan dari kurva standar. Sampel yang memiliki salinan DNA di bawah nilai LoD dapat dinyatakan positif. Materi genetik kura-kura brazil terdeteksi pada seluruh sampel danau di Universitas Indonesia. Faktor lingkungan tidak berpengaruh dalam pendeteksian kura-kura brazil. Berdasarkan hasil yang diperoleh, eDNA berpotensi kuat dalam pemantauan spesies invasif baru dan dapat digunakan untuk pengendalian lebih lanjut.

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The Red-eared Slider (Trachemys scripta elegans) is an invasive species from South America and has developed in various countries due to the release of unwanted pets which can have negative impacts on native species. This research was conducted to detect the presence of the species Trachemys scripta elegans in six lakes at Universitas Indonesia, Depok, West Java. The species detection was carried out using environmental DNA (eDNA) and quantitative PCR (qPCR) method with 12S rRNA target gene. The genetic material was isolated using the PCI method and 12S rRNA target gene were quantified using qPCR method. The presence of the target gene in the sample was determined by the LoD and LoQ values obtained from the standard curve. Samples that have DNA copies under the LoD value can be tested positive.Genetic material of the Red-eared Slider was detected in the all lake samples of University of Indonesia. Environmental factors have no effect on the detection of the Red-eared Slider. Based on the results, eDNA has strong potential in monitoring the new invasive species and can be used for further control."

"Kura-kura brazil (Trachemys scripta elegans) merupakan spesies asal Amerika Selatan dan salah satu spesies asing invasif yang berdampak buruk untuk spesies asli. Proses invasi spesies tersebut di Indonesia adalah melalui jalur perdagangan dan populer sebagai hewan peliharaan. Pendeteksian kehadiran spesies asing menjadi penting dalam proses pengendalian spesies sebelum menjadi invasif. Ekosistem perairan urban seperti situ di Universitas Indonesia merupakan wilayah yang umum ditemukan spesies asing. Tujuan penelitian adalah mendeteksi keberadaan kura-kura brazil di enam situ Universitas Indonesia menggunakan sampel eDNA yang diamplifikasi menggunakan primer spesifik Cytochrome b dan dikuantifikasi menggunakan qPCR. Nilai LoD dan LoQ ditentukan melalui kurva standar untuk menentukan keberadaan DNA kura-kura brazil pada 193 sampel. DNA Kura-kura brazil terdeteksi pada sampel dari seluruh situ di Universitas Indonesia pada tahun 2021 dan 2022 Faktor lingkungan tidak memiliki pengaruh yang signifikan (sig. > 0,05) terhadap konsentrasi eDNA, tetapi kura-kura brazil ditemukan pada situ dengan air yang tenang dengan suhu 27 - 33°C. Berdasarkan hasil tersebut eDNA dapat digunakan untuk monitoring keberadaan spesies asing invasif kura-kura brazil di ekosistem urban.

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The red-eared sliders (Trachemys scripta elegans) is a species from Southern America and one of the invasive alien species that has a negative impact on native species. The invasion process in Indonesia is through trade routes and is popular as a pet. Detecting the presence of alien species is important in the process of controlling species before they become invasive. Urban water ecosystems such as the one at the University of Indonesia are areas that are commonly found by alien species. The aim of the study was to detect the presence of Brazilian turtles in six ponds at the University of Indonesia using eDNA samples amplified using Cytochrome b specific primers and quantified using qPCR. LoD and LoQ values ​​were determined using standard curves to determine the presence of Brazilian tortoise DNA in 193 samples. The red-eared sliders’ DNA was detected in samples from all ponds at the University of Indonesia in 2021 and 2022. Environmental factors did not have a significant effect (sig. > 0.05) on eDNA concentrations, but the red-eared slider were found in situ with contaminated water. quiet with temperature 27 - 33°C. The results based on the eDNA can be used to monitor the presence of an invasive alien species in urban ecosystems."

"Kura-kura brazil (Trachemys scripta elegans) merupakan salah satu spesies invasif perairan tawar. Keberadaan spesies tersebut membawa dampak buruk bagi ekosistem sehingga perlu dieradiksi dengan cara pendeteksian dini. Pendeteksian dini suatu spesies akuatik dapat dilakukan menggunakan pendekatan molekular yang efektif dan non-invasif, yaitu metode environmental DNA dan quantitative PCR. Penelitian dilakukan untuk mendeteksi keberadaan kura-kura brazil di enam situ di Universitas Indonesia, Depok, Indonesia. Metode yang digunakan dalam peneitian ini adalah isolasi dengan PCI, amplifikasi dengan qPCR secara triplikat menggunakan primer COI dari gen mitokondria. Berdasarkan pengujian limit of detection (LOD) dan limit of quantification (LOQ) yang ditentukan dari kurva standar memiliki LOD sebesar 220.164 salinan DNA/reaksi dan LOQ sebesar 667.163 salinan DNA/reaksi. Keberadaan kura-kura brazil terdeteksi di lima situ pada tahun 2021 dan enam situ pada tahun 2022. Tidak ditemukan pengaruh faktor lingkungan terhadap keberadaan kura-kura brazil yang ditentukan berdasarkan ANOVA Satu Arah dengan nilai p > 0,05. Keberadaan Kura-kura brazil di Universitas Indonesia dapat dideteksi menggunakan eDNA dan digunakan sebagai kegiatan pemantauan dan eradikasi spesies asing invasif di ekosistem perairan urban.

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Red-eared slider (Trachemys scripta elegans) is one of the most invasive freshwater species. The existence of this species affects their non-native ecosystem in a negative manner, that ideally it should be eradicated by conducting an early detection of the ecosystem. Early detection of an aquatic species can be done by using the environmental DNA and quantitative PCR methods, as both use effective molecular and non-invasive approach. This study was conducted to detect the presence of red-eared slider within the ecosystem of 6 ponds located at Universitas Indonesia, Depok, Indonesia. The methods that are used in the study covered isolation with PCI, amplification with qPCR in triplicate using primer COI from the mitochondria gen. Limit of detection (LOD) and limit of quantification (LOQ) were then examined by referring to the standard curve, LOD held the value of 220,164 of DNA copies/reaction, while LOQ held the value of 667,163 of DNA copies/reaction. The presence of red-eared slider was then proven in the ponds within the ecosystem; 5 in 2021 and 6 in 2022. The influence between the presence of red-eared slider and pH was not found, the conclusion is backed with calculation using One Way ANOVA using p-value > 0,05. The presence of red-eared slider in Universitas Indonesia can be detected using eDNA, which later can be utilized as a tool to observe and eradicate the foreign species within the urban water ecosystem"