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

Carotenoid pigment content and skin microscopic appearance of cultured Ajamaru Rainbowfish (Melanotaenia ajamaruensis)

The color of Ajamaru Rainbowfish (Melanotaenia ajamaruensis), which is adapted to the tank culture tends to decrease in brightness and become dull. It is necessary to improve the quality of color, especially through the diet. This study was conducted to determine the effect of the supplementation of carotenoid on the diet on the amount of carotenoid pigments and the microscopic appearance of the skin, fins, and scales of the Ajamaru rainbowfish. The research used a factorial design, consisting of two factors and three replications. The Factors were three different diets (carotenoid-free diet, astaxanthin, and lutein diet) as factor 1, and gender (male and female) as factor 2. The observed data were carotenoid content in fins, skin, and flesh; the microscopic appearance of fins and scales was measured at the end of the study. The results showed that the supplementation of astaxanthin and lutein to the feed increased the content of carotenoid pigments in the fins and scales of the Ajamaru rainbowfish; the highest values in the fins and skin of the male fish were 202.90 and 8.53 ppm, respectively. The effect of astaxanthin and lutein supplementation on color can be observed microscopically in the tissue of scales and fins

PHYTOCHEMICAL COMPOSITION AND ANTICANCER ACTIVITY OF SEAWEEDS ULVA LACTUCA AND EUCHEUMA COTTONII AGAINST BREAST MCF-7 AND COLON HCT-116 CELLS

Objective: this study is aimed to develop marine resources which is focused on the determination of  phytochemical composition and exploration of seaweeds Ulva Lactuca and Eucheuma cotonii, as a potential  anti-breast cancer and anti-colorectal cancer agents. Methods: Seaweeds Ulva Lactuca collected from Parangtritis beach, Yogyakarta, Central Java, Indonesia. Whereas Eucheuma cottonii collected from Salemo island, South Sulawesi, Indonesia. Seaweeds U. lactuca and E. cottonii were macerated in organic solvents, n-hexane, chloroform, ethyl acetate and ethanol, respectively. After maceration for three days, the mixture was filtered, the filtrate was concentrated by rotary evaporator. The concentrated extract of n-hexane, ethyl acetate, ethanol and chloroform were then analyzed by thin layer chromatography. Phytochemical test of the concentrated extract were conducted to identify the metabolites containing in the seaweeds. Furthermore, the cytotoxic activity of the n-hexane, ethyl acetate, ethanol and chloroform extract of Ulva Lactuca and Eucheuma cotonii were evaluated as a growth inhibitor of breast MCF-7 and colorectal HCT-116 cancer cells by MTT cell proliferation assay. Results: Phytochemical test  for the  concentrated extracts of Ulva Lactuca showed the positive result for metabolites of  steroids, glycosides, flavonoid,and tannin. While  the concentrated extracts of E. cottonii showed positive result for metabolites of steroids, glycosides, and flavonoid. Both concentrated extracts of Ulva lactuca and Eucheuma cotonii exhibited anticancer activity against breast MCF-7 and colorectal HCT-116 cells with IC50 ranging of  21 µg/mL  to  99 µg/mL . Conclusion: Our results clearly demonstrate seaweeds Ulva Lactuca and Eucheuma cotonii as a promising candidates for the  new anti-breast and anti-colorectal cancer agents. Keywords: Phytochemistry, Ulva Lactuca, Eucheuma cotonii , anticancer, breast MCF-7, colorectal HCT-11

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

Biotechnological Potential of Bacteria Isolated from the Sea Cucumber Holothuria leucospilota and Stichopus vastus from Lampung, Indonesia

In order to minimize re-discovery of already known anti-infective compounds, we focused our screening approach on understudied, almost untapped marine environments including marine invertebrates and their associated bacteria. Therefore, two sea cucumber species, Holothuria leucospilota and Stichopus vastus, were collected from Lampung (Indonesia), and 127 bacterial strains were identified by partial 16S rRNA-gene sequencing analysis and compared with the NCBI database. In addition, the overall bacterial diversity from tissue samples of the sea cucumbers H. leucospilota and S. vastus was analyzed using the cultivation-independent Illumina MiSEQ analysis. Selected bacterial isolates were grown to high densities and the extracted biomass was tested against a selection of bacteria and fungi as well as the hepatitis C virus (HCV). Identification of putative bioactive bacterial-derived compounds were performed by analyzing the accurate mass of the precursor/parent ions (MS1) as well as product/daughter ions (MS2) using high resolution mass spectrometry (HRMS) analysis of all active fractions. With this attempt we were able to identify 23 putatively known and two previously unidentified precursor ions. Moreover, through 16S rRNA-gene sequencing we were able to identify putatively novel bacterial species from the phyla Actinobacteria, Proteobacteria and also Firmicutes. Our findings suggest that sea cucumbers like H. leucospilota and S. vastus are promising sources for the isolation of novel bacterial species that produce compounds with potentially high biotechnological potential

Isolasi senyawa triperpenoid dari daun Artocarpus Elcaticus (Bendo)

Isolasi kandungan senyawa triterperiold dari daun ArLocarpus eiasticus dilakukan dengan sokletasi menggunakan pelarut n-heksan. Metode pemisahan yang digunakan adalah kromatografi kolom sistem gradien menggunakan pelarut heksan etil asetat dan face padatnya silika gel. Terhadap fraksi ke-3 dilakukan pemurnian dengan metanq diperoleh kristal berwarna putih dengan titik leleh 84-86° C. Dari pengujian spektroskopi UV dan IR didapat bahwa senyawa hasil isolasi mempunyai gugus fungsi OH dan ikatan C=C tak terkonjugasi. Spektroskopi massa menunjukkan pada m/e : 476. Dalam penelitian ini struktur molekul belum dapat ditentukan, perlu dilakukan penelitian le An Isolation of Triterpenoid compound from Artocarpus elasticus leaves was done by soxhlet using n-heksan as a solvent. The separation method applied to was coulomn chromatography with gradien system use solvent n-heksan - etil asetat and the solid fase was silica gel. The third fraction was purified using methanoland white crystalin with melting point of 84-86 C was obtained. From ultra violet and infra merah spectroscopy analysis it can be concluded that the compound resulted from the isolation has functional group of OH and non conjugated C=C bond .Mass spectroscopy resulted in that m/e: 476.In This research, the molecular structure can not be determined, because it still needs further research to be done. bih lanjut

BIODEGRADASI FENANTREN OLEH BAKTERI LAUT Pseudomonas sp KalP3b22 ASAL KUMAI KALIMANTAN TENGAH

The Biodegradation of Phenanthren by Marine Bacteria Pseudomonas sp KalP3b22 from Kumai Central Borneo. The study of the potential marine bacteria that degrade Polycyclic Aromatic Hydrocarbons (PAHs) was already done among several marine bacterium isolated from the contaminated sea water collected in Kumai Port area. The biodegradation rate of phenanthrene by selected bacteria Pseudomonas sp Kalp-3b22 was 59,5% after 29 days cultivation. The structural determination of degradation product was deduced as 1-naphtalenol. Keywords: Phenanthrene, biodegradation, Pseudomonas sp

THE ANTIBACTERIAL COMPOUND COLLISMICIN A DERIVED FROM MARINE <i>Streptomyces</i> sp Q-629K

In our course of screening for secondary metabolite derived from marine bacterial, we isolate the antimicrobial compound collysmicin A from the ethyl acetate extract of Streptomyces sp Q-629K. Separation of this compound was carried out by silica gel open column chromatography. Purification of an active compound was done using HPLC C18 with acetonitril-water system. Determination of chemical structure was done by 1H, 13C NMR and LC-MS analysis. Collysmicin A was contained in fraction 3, fraction 7.2 and fraction 8.7. The antimicrobial assayed from purified compound Fr.8.7 gave diameter inhibition approximately 13 mm against S. aureus and 12 mm against B. subtilis .   Keywords: antimicrobial, collismycin A and marine Streptomyces s