IDENTIFIKASI SPESIES MENGGUNAKAN DNA BARCODING DALAM MENUNJANG BUDIDAYA DAN KONSERVASI TERIPANG DI PERAIRAN LAMPUNG

Teripang merupakan komoditas perikanan yang saat ini dibudidayakan dan dieksploitasi di perairan Lampung. Namun terdapat kesulitan dalam mengidentifikasi teripang karena kemiripan morfologis di antara spesies yang ada. Identifikasi yang baik berguna agar proses pembudidayaan dan konservasi dapat tepat sasaran. Penggunaan DNA barcoding dapat digunakan untuk mengidentifikasi jenis-jenis teripang yang ada, jarak genetik, dan keragaman genetik intra/inter spesies. Penelitian ini bertujuan untuk mengidentifikasi teripang di perairan Lampung dengan menggunakan sekuen DNA gen COI. Teripang diambil dengan menggunakan metode jelajah pada saat surut dan dengan scuba diving. Pengamatan DNA menggunakan primer universal ceF, pengeditan dan diurutkan dengan program Geneious ver 9 dan program BLAST. Konstruksi pohon filogenetik dilakukan dengan metode neighbor joining (NJ) pada model Kimura-2. Penelitian ini menunjukkan spesies teripang yang teridentifikasi adalah Holothuria leucospilota, H. atra, Stichopus vastus, dan S. horrens dengan jarak kesamaan 99%-100%. S. vastus dan S. horrens memiliki jarak genetik terendah dengan pengurangan yang tinggi. Rekonstruksi filogenetik memperlihatkan pengelompokkan spesies-spesies ke dalam genus Holothuria dan Stichopus. Stichopus sp. memiliki kesamaan morfologi yang tinggi sehingga kesalahan identifikasi sering terjadi. DNA barcoding dapat mengidentifikasi teripang secara cepat dan akurat sehingga pengelolaan teripang baik secara budidaya maupun pengambilan langsung di alam dapat berkelanjutan. Identifikasi spesies yang tepat menjadi kunci utama dalam upaya pembudidayaan dan konservasi teripang yang tepat sasaran dan berkelanjutan.Sea cucumbers is a highly valued fishery commodity that is currently cultivated and exploited in Lampung waters. However, differentiating a sea cucumber species from another is sometimes difficult due the morphological similarities between the species. Developing an accurate identification method is then critical to ensure successfull farming activities and conservation efforts of sea cucumbers. DNA barcoding could be used to accurately identify sea cucumber species, genetic distance, and genetic diversity between species. This study aimed to identify sea cucumbers existed in Lampung waters using DNA barcoding of the COI gene with ceF and ceR universal primers. Sea cucumbers are taken using the cruising method at low tide and by scuba diving. The DNA sequence was then edited and aligmented using the Geneious ver.9 program and analyzed using the BLAST program. Phylogenetic tree construction was carried out using the neighbor joining (NJ) method on the Kimura-2 model. This study showed that the identified species of sea cucumbers were Holothuria leucospilota, H. atra, Stichopus vastus, and S. horrens with a similarity distance of 99%-100%. S. vastus and S. horrens have the lowest genetic range. Phylogenetic reconstruction shows the classification of species into the genus Holothuria and Stichopus. Stichopus sp. have high morphological similarities within the same genus which often lead to species misidentification. DNA barcoding can identify sea cucumbers quickly and accurately. This method allows the identification of the right sea cucumber species which is the main key in the effort to cultivate and conserve targeted and sustainable sea cucumbers

ACUTE ORAL TOXICITY ASSESSMENT OF THE ETHANOL EXTRACT OF HOLOTHURIA ATRA IN MICE

Objective: This study aimed to evaluate the toxicological potential of the ethanol extract of Holothuria atra through the acute oral toxicity – acute toxic class method. Methods: The sample was immersed in ethanol for 72 h at room temperature and repeated 3 times. The extracts were evaporated using a vacuum rotary evaporator. The identification of compounds in the ethanol extract of H. atra was carried out using liquid chromatography–mass spectrometry (LCMS) analysis. The acute toxicity test was examined the effects of treating male mice with the ethanol extract of H. atra at 300 and 2000 mg/kg by oral administration for 14 days. On the past day of the toxicity test, liver of all experimental animals was taken for histopathological testing. Results: LCMS analysis showed that the ethanol extract of H. atra is contained polar compounds (chlorogenic acid, coumaric acid, a glycosaminoglycan, and holothurin) and non-polar compounds (fatty acids). Acute toxicity study was performed at a dose of 300 and 2000 mg/kg for 14 consecutive days. No deaths or behavioral changes were observed during the administration of both doses. Histopathological test results on the liver showed a few changes at doses of 2000 mg/kg. Conclusions: The LD50 is equal to 5000 mg/kg and the ethanol extracts of H. atra can be classified as practically nontoxic. However, further studies are required to proceed to clinical studies in humans

Pengaruh Nutrisi Dan Suhu Terhadap Selektivitas Potensi Antibakteri Dari Bakteri Yang Berasosiasi Dengan Spons

roduction of pharmacological activity by marine microorganism is strongly influenced by nutrition and environmental conditions. In this study would  discuss about the influence of several type of media to the production of antibacterial agent by sponge-. associated microorganisms. About 3 sponges tissue Theonella sp, Callispongia sp. and Lithistide sp. collected from Seribu Island will be used for the host of associated microorganism. Agar medium used for isolation were M1 that contained amylum,  yeast extract and peptone, M2 (10% marine broth media) contained yeast extract and peptone, M3 only sea water without adding any nutrients. Beside the nutrient variation,  heat sock treatment at 50oC toward the sponge solution also apply to this study. The bacterial isolation data indicated that bacterial density in (CFU/100µL) of  Theonella sp, Callispongia sp. and Lithistide sp. were minimum when spreading in M3 medium with heat sock treatment. This data showed that limiting in nutrient and heating could increasing bacterial selectivity. The antibacterial activity capability of bacterial strains isolated using M1, M2 and M3 respectively in range were 81,8-90,9%; 50-87,5%  and; 66,7 -100%. This results showed that less nutrient of media will rise the number of antibacterial activity strains,and decreasing of bacterial density. This study reported that the minimum nutrient of isolation media and heat shock treatment could be used for selecting the antibacterial strains of sponge associated bacteria.   Aktifitas farmakologi yang dihasilkan oleh mikroorganisme laut sangat dipengaruhi oleh nutrisi dan kondisi lingkungan. Hal tersebut mendorong untuk digali lebih dalam tentang aspek-aspek yang mempengaruhi seleksi mikroba potensial pada spons. Metode isolasi mikroba dari jaringan spons menjadi kunci dalam menguak potensi mikroba simbionnya. Dalam penelitian ini akan membahas pengaruh berbagai media isolasi bakteri yang berasosiasi dengan spons asal Kep. Seribu. Terhadap 3 specimen spons Theonella sp., Callispongia sp. dan Lithistide sp. dilakukan isolasi bakteri dengan metode direct sampling menggunakan media M1, M2 dan M3. Media M1 mengandung nutrisi antara lain amilum, ekstrak khamir dan pepton, sedangkan media M2 mengandung sumber nutrisi ekstrak khamir dan pepton dan M3 hanya media agar dan air laut. Selain variasi nutrient dalam media, perlakuan pemanasan pada suhu 50oC juga akan dilakukan terhadap larutan sampel spons sebelum dilakukan penyebaran pada media isolasi. Hasil isolasi bakteri yang diisolasi spons Theonella sp, Callispongia sp. dan Lithistide sp. menunjukkan bahwa kepadatan minimum diperoleh dengan menggunakan media M3 dengan perlakuan pemanansan. Dari data isolasi bakteri menunjukkan bahwa selain kandungan nutrient yang minimum, perlakuan pemanasan akan menurunkan kepadatan jumlah bakteri yang tumbuh, sehingga pemanasan merupakan salah satu cara dalam seleksi isolasi bakteri yang berasosiasi dengan spons. Hasil analisis aktivitas antibakteri menunjukkan bahwa persentase strain-strain bakteri yang aktif terhadap antimikroba Vibrio eltor, Eschericia coli dan Bacillus subtilis dengan variasi media M1 berkisar antara 81,8-90,9%;  M2 berkisar 50-87,5% dan M3 berkisar 66,7-100%. Dari data tersebut disimpulkan bahwa semakin sedikitnya nutrisi media isolasi maka semakin tingginya mikroba-mikroba potensial penghasil antibiotik. Media M3 merupakan media yang selektif untuk isolasi mikroba potensial dari spons, terbukti dengan tingginya prosentase bakteri yang aktif dan berkurangnya jumlah koloni yang tumbuh

Bioassay-Guided Isolation of an Antibacterial Compound from the Indonesian Soft Coral

With the aim of searching for new antibacterial compound from marine soft corals, the investigation had been conducted on antibacterial activity of an extract from soft corals Sarcophyton trocheliophorum in the n-hexane, ethyl acetate, n-butanol, and aqueous fractions. The antibacterial activity was tested against two Gram-positive bacteria, viz. Bacillus subtilis (ATCC 6633) and Staphylococcus aureus (ATCC 25923), and two Gram-negative bacteria, viz. Escherichia coli (ATCC 25922) and Vibrio cholerae (ATCC 14035) using the agar disc diffusion assay. Among them, the n-hexane fraction was the most active against three tested bacteria, viz. Bacillus subtilis, Staphylococcus aureus and Vibrio cholerae at the concentration 125 µg/ml, with inhibition zone 14.2, 18.2, 13.8 mm, respectively. Isolation and purification of the active component from the n-hexane fraction led to a known cembranoid-type diterpene, sarcophytoxide. The chemical structure of the isolated compound was determined by IR, MS and NMR, as well as compared to data from the literature. Sarcophytoxide showed moderate activity against B. subtilis, S. aureus and V. cholerae, with a minimum inhibitory concentration (MIC) of 125, 100, 125 mg/ml, respectively

SOFT CORAL (Sinularia sp.) EXTRACTS WITH ANTIBACTERIAL ACTIVITY

There is a growing need for new antibacterial agents, in particular because many antibiotics are becoming ineffective due to the emergence of resistant bacterial strains. Soft corals of the Genus Sinularia, Family Alcyoniidae, have potential as a source of terpenoid and steroid compounds with antibacterial activity. These corals may vary in external morphology (shape, colour, size).The aim of this research was to identify extracted fractions with high antibacterial activity. Sinularia sp. specimens were extracted, fractionated based on solvent polarity, and tested for antibacterial bioactivity against pathogenic bacteria (Escherichia coli and Staphylococcus aureus). Antibacterial activity of the three fractions varied in strength. The dichloromethane fraction showed strong antibacterial activity, inhibiting S. aureus and E. coli growth at a concentration of 1 mg ml-1, while the ethyl acetate and ethanol fractions were effective at 10 mg ml-1 and 100 mg ml-1, respectively

Secondary metabolites and their biological activities in Indonesian soft coral of the genus Lobophytum

AbstractObjectiveTo investigate the antioxidant, antibacterial, antimalarial activities and cytotoxicity of the n-hexane, ethyl acetate, n-butanol, and aqueous fractions from a crude extract of Lobophytum sp.MethodsThis organism was collected from the Selayar Islands (South Sulawesi). The antioxidant activity was performed by the 1,1-diphenyl-2-picryl hydrazyl radical scavenging method. All fractions from the crude extract of Lobophytum sp. were examined for their cytotoxicity using the brine shrimp lethality bioassay and heme polymerization inhibitory activity assay for antimalarial activity.ResultsIt was found that the ethyl acetate, n-butanol and aqueous fractions exhibited heme polymerization inhibitory activity with IC50 values of 11.7, 14.3 and 12.0 μg/mL, respectively, while the n-butanol fraction showed moderate antioxidant activity and cytotoxicity with IC50 values of 150.00 and 92.74 μg/mL, respectively.ConclusionsThis study provides information on antioxidant, antibacterial and antimalarial activities as well as the cytotoxicity of all fractions from the crude extract of Lobophytum sp. This is a new report of antimalarial substances derived from Lobophytum sp

AKTIVITAS ANTIOKSIDAN DARI METABOLIT SEKUNDER KAPANG ENDOFIT MANGROVE Aegiceras corniculatum

ABSTRACTMangrove endophytic fungi potentially produce secondary metabolites such as antioxidant. Antioxidants are compounds that are widely applied in the industry as an ingredient in the manufacture of products in the food and health sector. In this study, we investigated the antioxidant  potential of secondary metabolites from fungal endophytic of mangrove species Aegiceras corniculatum. A total of eight endophytic fungi were successfully isolated, two isolates from leaf, three isolates from fruit and three isolates from twig.  Between the isolates obtained, only one isolated was active as the antioxidant with IC50 value 19.28 µL/mL eventhough still lower than the standard ascorbic acid (6.08 µL/mL). The results of chemical identification using GC-MS showed several chemical compounds that have antioxidant activity including phenol,3,5-bis(1,1-dimethylethyl), hexadecanoic acid, hexadecanoic acid methyl ester, malic acid, N-aminopyrrolidine,  9-octadecanoic acid, methyl ester (E), benzeneethanol, 4-hydroxy, 1,2-benzenedicarboxylic acid, d-tyrosine, bis(2-methylpropyl) ester 1-nonadecene dan heneicosane.  The selected fungal endophytic isolated were identified using molecular Internal Transcribed Spacer (ITS) marker and has a high taxonomy similarity with Microdochium sp.Keywords: Aegiceras corniculatum, antioxidant, endophytic fungi, mangrove, Microdochium sp ABSTRAKKapang endofit dari mangrove memiliki potensi dalam menghasilkan metabolit sekunder seperti antioksidan. Antioksidan merupakan senyawa yang banyak diaplikasikan dalam industri sebagai  bahan untuk pembuatan produk di bidang pangan maupun kesehatan. Penelitian ini bertujuan untuk  mengetahui potensi antioksidan kapang endofit dari mangrove species Aegiceras  corniculatum. Sebanyak 8 isolat kapang endofit berhasil diisolasi, yaitu 2 isolat berasal dari daun, 3 isolat berasal dari buah dan 3 isolat berasal dari ranting.  Di antara 8 isolat tersebut, hanya 1 isolat yang dihanyakan aktif sebagai antioksidan dengan nilai IC50  sebesar 19,28 µL/mL, meskipun nilai ini masih lebih rendah dibandingkan vitamin C yaitu sebesar 6,08 µL/mL. Hasil identifikasi kimia dengan menggunakan GC-MS menunjukkan beberapa senyawa kimia yang memiliki aktivitas aktioksidan di antaranya phenol,3,5-bis(1,1-dimethylethyl), hexadecanoic acid, hexadecanoic acid methyl ester, malic acid, N-aminopyrrolidine,  9-octadecanoic acid, methyl ester (E), benzeneethanol, 4-hydroxy, 1,2-benzenedicarboxylic acid, d-tyrosine, bis(2-methylpropyl) ester 1-nonadecene dan heneicosane. Isolat kapang endofit diidentifikasi dengan penanda Internal Transcribed Spacer (ITS) dan memiliki kemiripan tertinggi dengan Microdochium sp.Kata kunci: Aegiceras corniculatum, antioksidan, kapang endofit, mangrove, Microdochium s

Chemical Composition, Antimicrobial, Cytotoxic and Antiplasmodial Activities of Three Sponges from Buton Islands, Indonesia

GC-MS analysis of the crude extracts of three different species of Indonesian marine sponges has been carried out for identification of bioactive compounds. The GC-MS analysis from Haliclona (Gellius) sp., Lamellodysidea herbacea, and Spheciospongia inconstans revealed the presence of 23, 21, 19 various compounds, respectively and mainly sterols and fatty acids. All the sponge species has been evaluated for antimicrobial activities, cytotoxicity using brine shrimp lethality bioassay and heme polymerization inhibitory activity assay for antiplasmodial activity. In this study, all the sponge species showed antimicrobial activities against at leastone of the test strains. Among them, the extract of sponge Lamellodysidea herbacea displayed activity against two Gram-positive bacteria (S. aureus and B. subtilis) and the Gram-negative bacteria V. cholerae, with inhibition zones of 10.3, 9.2 and 9.5 mm, respectively. The sponge Haliclona (Gellius) sp., showed significant activity against fungal pathogen C. albicans. The sponge Haliclona (Gellius) sp., displayed the ability to inhibit heme polymerization indicating an anti-Plasmodium function and also showed potent cytotoxic activity against the brine shrimp Artemia sp.   Keywords: GC-MS analysis, antimicrobial, sponges,bioactiv