Antibacterial Screening Activity of Several Sponges Against Staphylococcus aureus, Escherichia coli, Staphylococcus saprophyticus, dan Pseudomonas aeruginosa

Sponge samples collected around Manado waters were obtained 30 species and their crude extracted have been tested in vitro for their activity in inhibiting bacterial growth. Based on the results of antibacterial screening in 30 sponge extracts, there were 23 sponge extracts which had bioactivity in inhibiting the growth of S. aureus, E. coli, S. saprophyticus and P. aeruginosa, Sponge extract No. 43 (of 30 sponge extracts tested) was the most active in inhibiting bacterial growth and had the widest inhibition zone diameter.Keywords: screening, sponge, crude extract, antibacterial, ManadoABSTRAKSampel spons dikoleksi di sekitar Perairan Manado sebanyak 30 jenis/spesies, dimana ekstrak kasarnya telah diuji secara in vitro aktivitasnya dalam menghambat pertumbuhan bakteri. Berdasarkan hasil skrining antibakteri pada 30 ekstrak spons didapatkan hasil ada 23 ekstrak spons yang mempunyai bioaktivitas dalam menghambat pertumbuhan bakteri S. aureus, E. coli, S. saprophyticus dan P. aeruginosa, Ekstrak spons No. 3 (dari 30 ekstrak spons yang diuji) adalah yang paling aktif dalam menghambat pertumbuhan bakteri dan memiliki diameter zona hambat yang paling lebar.Kata kunci: skrining, spons, ekstrak kasar, antibakteri, Manado

The Use Of Marine Sponge Crude Extract To Improve The Resistance Of Tilapia Fish (Oreochromis niloticus) To Streptococcus agalactiae Infections

This study aims to examine the effect of the use of crude marine sponge extract on the resistance of streptococcus agalactiae infection in tilapia, Oreochromis niloticus, and to establish the effective dose of crude sponge extract in improving the immune system and the growth of the fish. The sponge used in the study was Cribrochalina sp. taken from Malalayang waters, Manado. The fish were taken from Freshwater Aquaculture Center, Tatelu. The fish were acclimatized for a week. After being acclimatized the fish were given feed added with sponge crude extract as a treatment with different concentrations of 20 g, 40 g and 60 g / Kg of feed for 14 days as much as 5% / body weight / day with the frequency of feeding twice a day at 10:00 am and at 5:00 p.m. After being treated, the fish was challenged with S. agalactiae. The data collected consisted of tilapia resistance, Total Leukocyte Count (TLC) as immune parameters and absolute growth. The results showed that the addition of crude extracts of Cribrochalina sp. into feed can increase TLC and growth of tilapia (p <0.05). The best results were achieved in fish fed with the addition of sponge crude extract of 40 g/kg feed. The survival rate of tilapia fed with treatment diet then challenged with pathogenic bacteria S. agalactiae showed the best results (100% survival rate) compared to controls (75%). In conclusion, feeding with a crude extract of Cribrochalina sp. has the potential to increase the immune system and growth of tilapia.Keywords: Crude Extract, Marine Sponges, Cribrochalina sp., Tilapia, Resistance, Streptococcus agalactiaeABSTRAKPenelitian ini bertujuan untuk mengkaji pengaruh penggunaan ekstrak kasar spons laut terhadap resistensi ikan nila dalam menghadapi serangan Streptococcus agalactiae, mengidentifikasi spons yang digunakan, serta mengukur pengaruh serta menetapkan dosis pemberian ekstrak kasar spons untuk meningkatkan sistem imun dan pengaruhnya terhadap pertumbuhan ikan nila. Spons yang digunakan dalam penelitian adalah spons Cribrochalina sp. yang diambil dari perairan Malalayang. Ikan uji diambil dari Balai Budidaya Air Tawar Tatelu, Provinsi Sulawesi Utara. Ikan diaklimatisasi selama seminggu. Setelah diaklimatisasi ikan diberi pakan yang ditambahkan dengan ekstrak kasar spons sebagai perlakuan dengan konsentrasi berbeda yaitu 0 g, 20 g, 40 g dan 60 g/kg pakan selama 14 hari sebanyak 5%/berat tubuh/hari dengan frekuensi pemberian pakan dua kali sehari yaitu jam 10.00 pagi dan jam 17.00 sore. Setelah diberi perlakuan, ikan diuji tantang dengan bakteri S. agalactiae. Data yang dikumpulkan terdiri dari kelangsunganhidup ikan nila. Total leukosit sebagai parameter imun dan pertumbuhan mutlak. Hasil penelitian mendapatkan bahwa penambahan ekstrak kasar spons Cribrochalina sp. ke dalam pakan mampu meningkatkan total leukosit dan pertumbuhan ikan nila (p<0.05). Dimana hasil terbaik dicapai pada ikan yang diberi pakan dengan penambahan ekstrak kasar spons sebanyak 40 g/kg pakan. Kelangsungan hidup ikan nila yang diberi pakan perlakuan yang diuji tantang dengan bakteri patogen menunjukkan hasil yang paling baik (tingkat kelangsungan hidup 100%) dibandingkan dengan kontrol (75%). Sebagai kesimpulan bahwa pemberian pakan dengan ekstrak kasar spons Cribrochalina sp. berpotensi untuk meningkatkan sistem kekebalan tubuh dan pertumbuhan pada ikan nila.Kata Kunci: Ekstrak kasar, Spons laut, Cribrochalina sp., Tilapia, Resistensi, Streptococcus agalactiae

Cultivatingin situtriggers growth of previously uncultivated microorganisms via a growth initiation factor in nature

Most microorganisms resist cultivation under standard laboratory conditions. On the other hand, to cultivate microbes in a membrane-bound device incubated in nature (in situcultivation) is an effective approach. In the present study, we appliedin situcultivation to isolate diverse previously uncultivated marine sponge-associated microbes and comparatively analyzed this method’s efficiencies with those of the conventional method. Then, we attempted to clarify the key and unknown mechanism ofin situcultivation by focusing on growth triggering via growth initiation factor. We hypothesized that majority of environmental microorganisms are in nongrowing state and requiring “growth initiation factor” for the recovery and that can be provided from environments. Consequently, significantly more novel and diverse microbial types were isolated viain situcultivation than by standard direct plating (SDP). Next, the effect of the sponge extract on starvation recovery was compared between strains derived fromin situand SDP cultivation. Adding small amounts of the sponge extracts to the medium elevated the colony-formation efficiencies of thein situstrains at the starvation recovery step, while it showed no positive effect on that of SDP strains. Conversely, specific growth rates or carrying capacities of all tested strains were not positively affected. These results indicate that, 1) the sponge extract contains chemical compounds that facilitate starvation recovery, these substances selectively worked on thein situstrains, and 2) growth initiation factor in the sponge extract did not continuously promote growth activity but worked as triggers for regrowth (resuscitation from dormancy).ImportanceMost microbial species resist cultivation under laboratory condition. This is critical impediment for both academic and applied microbiology, and thus clarification of the mechanism of microbial uncultivability is highly demanded. Several evidences have been reported that to cultivate microbes in a membrane-bound device incubated in nature (in situcultivation) is an effective approach. However, the mechanism behind this approach has not been clarified. The present study shows the evidence that 1) initiating growth is a key for cultivating previously uncultivated microbes rather than simple growth promotion, and 2) growth initiation factor (signaling-like compounds) in natural environments stimulate microbial resuscitation from a nongrowing state. Since no study has focused on growth initiation for cultivation of previously uncultivated microorganisms, the discovery shown in the present study provides a new insight into microorganisms previously considered uncultivable and a microbial growth controlling system in nature.

Identification of Cytotoxicity of Marine Sponge Dendrilla Membranosa Against L929 Cell Line

Marine sponges are rich sources of pharmacological active compound. Marine sponge, Dendrilla membranosa was collected from the Vizhingam coast. The sponge extract was tested against eight human bacterial pathogens. The bioactive compounds present in marine sponge were determined by GC-MS analysis. The cytotoxic effect of the sponge was evaluated by using MTT assay. The extracts showed potent anti-bacterial activity against Staphylococcus aureus, Bacillus subtilis, Streptococcus pyogens. The bioactive compound present in marine sponge was identified by GC-MS analysis and the compounds are ethane, butane, formate pentane, Alpha d-galactopyranoside, 2-Ethylhexyl2-ethylhexanoate, Hexahydro hexitol, Styrene, Alpha-d glucopyranoside 1-pentyl-2-proppyl-1-octene, nonadecane, 1,2-benzenedicarboxylic acid, bis-2, Octanoic acid. The test material showed none cytotoxic response to fibroblasts cells. The results of present investigation revealed that, Dendrilla membranosa is a potential source of novel anticancer and antibacterial leads.

ANTIBACTERIAL ACTIVITY AS INHIBITORS PATHOGEN BACTERIAL ON POND SHRIMP OF EXTRACT MARINE BIOTA COLLECTED FROM MASPARI ISLAND, SOUTH SUMATERA, INDONESIA

Marine biota has variety of bioactive compound that is potential to be an antibacterial for shrimp ponds diseases. Some of marine biota has potential as antibacterial i.e. soft coral Sarcophyton sp., Sponge Aaptos sp., seaweeds Sargassum sp. and Halimeda sp. and mangroves Avicennia sp. and Rhizophora sp. species. This study is purposed to find the most potential marine biota as antibacterial. The research method was as follows; sampling, sample identification, extraction and antibacterial activity test. The results is soft coral and sponge extract Sarcophyton sp. has bioactivity against of shrimp bacteria pathogen such as Vibrio spp2. bacteria for around 6.3±0.1 mm, while Aaptos sp. has bioactivity against Vibrio spp1. bacteria for around 7.9+0.1 mm, Vibrio spp2. bacteria for around 7.2± 0.1 to 7.9±0.1 mm, Vibrio spp6. bacteria for around 7.5±0.2 mm, Escherichia coli for around 7.2±0.1 mm and Staphylococcus aureus for around 15.9±0.2 mm. Seaweed extract activity found only of Sargassum sp. which has antibacterial activity against for around 7.1±0.0 mm for Vibrio spp6. and mangrove species activity has Rhizophora sp. extract which has bioactivity against Vibrio spp4. were 7.3±0.1 mm and E. coli bacteria were 6.7±0.1 mm. The most potential marine biota as antibacterial is showed on sponge Aaptos sp.. with an inhibitory zone for around 15.9±0.2 mm (for S. aureus bacteria).

The Antibacterial Activity Of Several Sponges From The Waters Of Tasik Ria Against Escherichia coli And Staphylococcus aureus bacteria

The sponge is one of the sea organisms that has a prospect as a source of natural compounds including peptides, steroids, asetogenin, terpenoids, alkaloids, cyclic halide and nitrogen. This research was directed to obtain several species of sponges from the waters of Tasik Ria as well as testing the antibacterial activity of extracts from some of the sponge against the bacteria Escherichia coli and Staphylococcus aureus. From the identification, seven species of sponges were found, which consists of: Amphimedon sp., Axinosa sp., Aaptos sp., Theonella sp., Cribochalina sp., Hyrtios sp., and Lendenfeldia sp. The tests of antibacterial activity of the extracts from these sponges against test bacteria E. coli and S. aureus showed some positive results. Extract from Axinosa sp. sponge(16 mm) showed the strongest antibacterial activity on Escherichia coli bacteria. Followed by Hyrtios sp. extract (13.5 mm), Aaptos sp. extract (13 mm), Lendenfeldia sp. extract (13 mm) and Cribochalinai sp. extract(10.5 mm). While the the tests on Staphylococcus aureus bacteria showed that the strongest antibacterial activity was found from Axinosa sp. sponge extract (16.5 mm), followed by the extract from Aaptos sp. (15 mm), Lendenfeldia sp. extract (14.5 mm), Hyrtios sp. extract(13.5 mm) and Cribochalina sp. extract (11 mm).Keywords: Sponge, antibacterial, Escherichia coli, Staphylococcus aureus ABSTRAK Spons merupakan salah satu biota laut yang sangat prospektif sebagai sumber senyawa bahan-bahan alami antara lain peptide, terpenoid, steroid, asetogenin, alkaloid, halide siklik dan senyawa nitrogen. Penelitian ini diarahkan untuk mendapatkan beberapa spesies spons dari perairan Tasik Ria serta menguji aktivitas antibakteri dari beberapa ekstrak spons terhadap bakteri Escherichia coli dan bakteri Staphylococcus aureus. Hasil identifikasi spons ditemukan sebanyak tujuh spesies yang terdiri dari: Amphimedon sp., Axinosa sp., Aaptos sp., Theonella sp., Hyrtios sp., Cribochalina sp. dan Lendenfeldia sp.. Aktivitas antibakteri dari beberapa ekstrak spons terhadap bakteri uji E. coli dan S. aureus terdapat diameter zona hambat bervariasi yaitu bakteri Escherichia coli menunjukkan aktivitas antibakteri ekstrak spons terkuat pada spons Axinosa sp (16 mm), disusul ekstrak spons Hyrtios sp. (13,5 mm), ekstrak spons Aaptos sp. (13 mm), ekstrak spons Lendenfeldia sp. (13 mm) dan ekstrak spons Cribochalinai sp. (10,5 mm). Sedangkan pada bakteri Staphylococcus aureus menunjukkan aktivitas antibakteri ekstrak spons terkuat yaitu: ekstrak spons Axinosa sp. (16,5 mm), disusul ekstrak spons Aaptos sp. (15 mm), ekstrak spons Lendenfeldia sp. (14,5 mm), ekstrak spons Hyrtios sp. (13,5 mm) dan ekstrak spons Cribochalina sp.(11mm).Kata Kunci : Spons, Antibakteri, Escherichia coli, Staphylococcus aureus

Test of Larvacide Activity from Some Sponge Extracts to Aedes aegypti Larvae

Marine organisms have been known produce certain compounds those could lead for medicine purposes. Sponges are one and the most studied for this aim. oneof the important biological activities which expected from sponge are larvacide activity. The aims this research was to test the larvacide activity aagainst Aedes aegypti larvae from some of sponge extract. Sponge samples were taken from Malalayang Waters, (N 01 ° 27'37 "E 124 ° 47'30") on November 2014 with the depth varies from 2-15 m with SCUBA. The extraction, preparation of the larvae and activity testing was performed on Biomolecular and Marine Pharmacy Laboratory Faculty Fisheries and Marine Science. The sponge samples were cutted and soaked in 95% Ethanol for over night with 3 repetitions to obtain ethanolic extracts. The extract were filtered and evaporated using freeze dryer then tested onto 10 instars 3 instars m larvae that had been previously maintained. the test was made in triplowith 24 hours observation. abate was used as positive control while negative control clean water was used.The test results showed that of 11 Sponge tested, 10 species showed larvacidal activity and marine sponge extract Tedania sp. has the highest activity compared to 9 extracts. As a suggestion of this research the further purification of Tedania sp. extract is needed to know the structure of active compound.Keywords: Aedes aegypti, Larvacide, Sponge extractABSTRAKOrganisme laut yang dapat dikembangkan menjadi bahan sediaan obat antara lain sponge, dan merupakan salah satu organisme laut yang banyak diteliti. Beberapa aktivitas biologis penting yang diharapkan dari ekstrak sponge salah satunya adalah aktivitas larvasida. Penelitian ini bertujuan untuk menguji aktivitas larvasida nyamuk Aedes aegypti dari beberapa ekstrak Sponge. Sampel sponge diambil di perairan Malalayang, tepatnya di koordinat N 01°27’37” E 124°47’30” pada bulan November 2014 di kedalaman 2-15 m. Sedangkan untuk tahap ekstraksi, penyiapan larva uji dan pengujian aktivitas larvasida di lakukan di laboratorium Biomolekular dan Farmasitika Laut program studi Ilmu Kelautan, FPIK UNSRAT. Dalam penelitian yang dilakukan, sampel diambil di perairan menggunakan peralatan SCUBA. Setelah itu diekstrak dengan larutan etanol 95% dan direndam selama 24 jam dan dilakukan 3 kali pengulagan untuk mendapatkan ekstrak etanolik. Sampel dikeringkan dengan menggunakan alat freeze dryer kemudian diujikan ke 10 ekor larva nyamuk fase instar 3 yang telah dipelihara sebelumnya. Pengujian dilakukan sebanyak 3 kali pengulangan dengan lama pengamatan 24 jam pengamatan. Sebagai kontrol positif digunakan bubuk abate yang biasa dijual di pasaran sedangkan kontrol negative atau tanpa perlakuan digunakan air bersih. Data hasil pengamatan diolah menggunakan Microsoft excel.Hasil pengujian menunjukkan bahwa Dari 11 Sponge yang diuji, 10 jenis menunjukan aktivitas larvasida dengan persentase mortalitas yang bervariasi dan ekstrak sponge laut Tedania sp. memiliki aktivitas tertinggi dibandingkan dengan 10 jenis ekstrak sponge lain dalam pengujian. Sebagai saran dalam penelitian ini yaitu Perlu dilakukan pemurnian lanjut ekstrak lebih lanjut dari ekstrak sponge Tedania sp. yaitu ke tahap partisi dan Perlu adanya variasi konsentrasi dalam pengujian.Kata Kunci : Aedes aegypti, Larvasida, ekstrak Spong