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Isolation and Screening of Endophytic Microbes from Morinda citrifolia and their Ability to Produce Anti-Microbial Substances

Microbiology Indonesia Vol 1, No 3 (2007): December 2007
Publisher : Indonesian Society for microbiology

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Abstract

Assaying for, and isolation and screening of endophytic microbes from the Morinda citrifolia plant and their ability to produce anti-microbial substances was carried out. Endophytic microbes are microorganisms that live asymptomatically within the living tissue of host plants. Microorganisms such as bacteria, fungi, and yeast can be associated with the host and produce secondary metabolites. These secondary metabolites may be enzymes and other bioactive substances with medicinal activity such as anti-arthritic, anti-cancer, and anti-microbial compounds. The aims of these experiments was to investigate the ability of endophytic microbes isolated from M. citrifolia to produce secondary metabolites which can act as anti-microbial agents. A direct seed inoculating technique was used by planting the plant sample onto the surface of Nutrient Agar and Potato-Dextrose Agar. Assessment of their ability to produce anti-microbial substances was conducted by growing the endophyte isolates in Muller Hinton Broth for bacterial isolates, and in Potato-Dextrose Yeast Broth for fungal isolates. The agar diffusion method using paper disk was applied to assay the anti-microbial activity of each substance. The results of the endophyte isolation in these experiments gave five bacterial isolates and eleven fungal isolates. All of the bacterial isolates showed a broad antimicrobial spectrum while ten of the fungal isolate demonstrated broad anti-microbial activity and four out of the ten fungal isolates had activity towards Candida albicans.

AKTIVITAS ANTIBAKTERI REBUSAN SECANG (Caesalpinia sappan L.) TERHADAP Salmonella thypii SECARA IN VIVO (Antibacterial Activity of Boiled Secang Extract (Caesalpinia Sappan L.) Againts Salmonella Typhii in Vivo)

Jurnal Agritech Fakultas Teknologi Pertanian UGM Vol 33, No 01 (2013)
Publisher : Fakultas Teknologi Pertanian, Universitas Gadjah Mada

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Abstract

One of the plant that has been used emperically for treatment was Caesalpinia sappan L“secang”. The bark of this plant contains metabolites with strong and effective antibaterial activity, namely tannin and brasilin. Two methods, namely A and B, were employed to perform the in vivo antibacterial study of this plant. In every method, there were six groups (K1-K6) which each of the group consisted of five mice. K1-K3 were the control groups consisting of control using healthy mice (K1), positive control in the presence of chloramphenicol (K2) and negative control (K3).  The test groups were K4-K6, mice in these group were treated with different concentrations of “secang” water extract at 15%, 30% and 75%, respectively.  The Salmonella thypii bacterial suspension was injected to the mice via intraperitonium adminsitration. In A method, treatment was given 2 hours after infecting the mice with the bacterial suspension, while in the B method, the treatment was given to the mice after 24 hours of bacterial infection. After 3 days of treatment, the intraperitonium fluid of the mice was taken under anaesthetic condition followed by qualitative and quantitative analysis of the biological sample obtained. Quantitative analysis via A method demonstrated that a decreased of 16.54%, 25.29 % and 25.12% in bacterial colony number was observed in the samples collected from K4, K5 and K6 respectively when compared to the respective controls.  B method showed smalled decreased (3.37 %,  9.83 % and 20.48 %) in the colony number counted. The qualitative analysis demonstrated that the isolated bacterial strain was Salmonella typhi. In conclusion, boiled water extract of secang could act as effective antibacterial agent because it inhibit the growth of Salmonella thypii under in vivo study conditions. Keywords:  Antibacterial, extract, secang, Salmonella thypii, in vivo ABSTRAK Salah satu tanaman yang secara empiris banyak digunakan untuk pengobatan adalah Caesalpinia sappan L. (Secang). Batang dari tanaman secang mengandung tanin dan brasilin, yang berkhasiat sebagai antibakteri. Uji antibakteri rebusan secang dilakukan secara in vivo menggunakan 2 metode yaitu metode A dan metode B. Setiap metode terdiri dari 6 kelompok dan setiap kelompok terdiri dari 5 ekor mencit (mus musculus) putih jantan galur DDY. Masing-masing kelompok terdiri dari kontrol normal (K1), kontrol positif kloramfenikol (K2), kontrol negatif (K3) dan kelompok yang diberi rebusan secang konsentrasi 15 % (K4), 30 % (K5) dan 75 % (K6). Suspensi bakteri Salmonella thypii diinfeksikan ke mencit secara intraperitonium. Pada metode A, pengobatan diberikan 2 jam setelah mencit diinfeksi bakteri, sedangkan pada metode B, pengobatan diberikan 24 jam setelah mencit diinfeksi bakteri. Setelah tiga hari pengobatan, mencit dianestesi dan cairan intraperitonium mencit diambil untuk dilakukan uji kuantitatif dan uji kualitatif. Hasil uji kuantitatif metode A menunjukkan terjadi penurunan jumlah koloni bakteri dalam cairan intraperitonium mencit pada kelompok 4 (K4), 5 (K5) dan 6 (K6) terhadap kontrol negatif (K3) sebesar 16,54 %; 25,29 % dan 25,12 %; sedangkan pada metode B sebesar 3,37 %; 9,83 % dan 20,48 %. Hasil uji kualitatif pada metode A dan metode B menunjukkan bakteri yang terisolasi dari cairan intraperitonium mencit adalah Salmonella thypii. Hasil penelitian menunjukkan rebusan secang dapat menghambat pertumbuhan Salmonella thypii secara in vivo. Kata kunci: Antibakteri, rebusan secang, Salmonella thypii, in vivo

Cytotoxic assay of endophytic fungus 1.2.11 secondary metabolites from Brucea javanica (L) Merr towards cancer cell in vitro

Medical Journal of Indonesia Vol 15, No 3 (2006): July-September
Publisher : Faculty of Medicine Universitas Indonesia

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Cytotoxic assay of secondary metabolite endophytic fungus 1.2.11 from Brucea javanica (L) Merr has been carried out. Brucea javanica fruit collected from Cianjur was used in this experiment. Cytotoxic assay was done on Raji, NS-1, HeLa and Vero cells. The observation was done for 24 hours and also for 48 hours. IC50 was calculated using the Rich and Muench theory. To observe the working mechanism of cytotoxic process, DNA staining with etidium bromide and acridine orange was conducted. The cytotoxic assay of endophytic fungi 1.2.11 showed an IC50 of 58.35 μg/ml, 88.39 μg/ml on Raji cell,; 162.09 μg/ml, 66.24 μg/ml on NS cell; 361.21 μg/ml, 219.97 μg/ml on HeLa cell; and lastly 1075.18 μg/ml, 656.82 μg/ml on Vero cell after 24 and 48 hour incubation respectively. The results of this study showed that secondary metabolite of endophytic fungus 1.2.11 has selective cytotoxic effect towards cancer cell and also showed that it might cause apoptosis in NS-1cell. (Med J Indones 2006; 15:137-44) Keywords: Brucea javanica (L.) Merr, endophytic microbe, Cytotoxic assay, endophytic isolate 1.2.11, apoptosis

Phytochemical Screening and Toxicological Evaluation Using Brine Shrimp Lethality Test (BSLT) of Some Fractions of Prasman Leaves (Eupatorium triplinerve V) Extract

Indonesian Journal of Cancer Chemoprevention Vol 2, No 1 (2011)
Publisher : Indonesian Research Gateway

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Abstract

Prasman leaves (Eupatororium triplinerve V) was well documented to have anti cancer benefit in Indonesian traditional medicine history. However, there were noscientific studies including toxicological assessment on the planextract.The purpose of this study was to investigate thetoxicological effect of some fractions of Prasman leavesmethanol extract. Phytochemical screening by the Farnsworth method on powder and some fraction of the methanol extractwere conducted followed by toxicitytest using the “Brine Shrimp Lethality” test (BSLT) method. In the current study results, the phytochemical screening showed the presence of flavonoid, saponin, coumarin, tannin, steroidandvolatileoil.LC50ofthen-hexanefraction238.660g/mL,ethylacetatefraction 24.420g/mL,andn3butanol64.100g/mL.

n-Butanolic fraction of endofitic fungi of Buah Makasar increases apoptotic effect of doxorubicin on MCF-7 cells

INDONESIAN JOURNAL OF PHARMACY Vol 20 No 1, 2009
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

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Abstract

Makassar fruit, Brucea javanica (L.) Merr., showed chemopreventive activity. Secondary metabolites come from B. javanica fruit, brucatol and bruceantine, induced cell differentiation and apoptosis on Leukemia cell, while quassinoid and its derivates acted as antitumor promoter. Butanolic fraction of supernatan of endofitic fungi 1.2.11 isolate fermentation which isolated from B. javanica fruit showed cytotoxicity toward several cancer cells. This fraction has been predicted contain secondary metabolites from B. javanica and has been identified as Bruceosin and Canthin-6-one derivates. Butanolic fraction (FB) of supernatan from endofitic fungi 1.3.11 isolate fermentation is predicted for having similiar cytotoxycity as active as 1.2.11 isolate. This research is aimed to explore cytotoxycity potention dan doxorubicin on MCF-7 breast cancer cell.Synergism of BF-doxorubicin combination detect from cell viability inhibition and apoptosis induction on MCF-7, a breast cancer cell lines which shows resistancy toward doxorubicin. Cell viability on single treatment of FB and doxorubicin and its combination were carried out by MTT assay to determine IC50 and combination index (CI). Apoptosis induction of FB, doxorubicin and its combination were carried out by ethidium bromideacridine orange DNA staining.n- Butanolic fraction and doxorubicin showed cell viability inhibition on MCF-7 cell with IC50 48 μg/mL and 148 nM, respectively. Both of FB and doxorubicin showed apoptosis induction on IC50. Combination of FBdoxorubicin showed synergism and increased apoptosis induction on MCF-7 cell.Key words: Brucea javanica, endofitic fungi, MCF-7 cell, synergism, doxorubicin.

UJI AKTIVITAS ANTIMIKROBA METABOLIT SEKUNDER KAPANG ENDOFIT TANAMAN TRENGGULI (Cassia fistula L) (Antimicrobial Activity Test of Secondary Metabolite of Endophytic Fungi from Cassia fistula L)

Jurnal Bahan Alam Indonesia Vol 6, No 2 (2007)
Publisher : Indonesian Research Gateway

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Abstract

Assay  for  secondary  metabolite  of  endophytic  fungi  from  Cassia  fistula  (L),  (Trengguli  Plant)  has been done. Endophytic fungi are microorganisms that live a symptomatically within the living tissue of host plants. Endophytic fungi associate with plant; they can help the metabolism of host plant and produce potential secondary metabolite. The purpose of this experiment was to investigate endophytic microbe  that  could  produced  antimicrobial  agents.  Isolation  of  endophytic  fungi  from  branch  of  Cassia fistula (L), (trengguli plant) using direct seed methods were carried out. The plants sample was put on the surface of Potato Dextrose Agar. Assessments of their ability to produce anti microbial substances were conducted by agar diffusion method using paper disc. The results showed that 10 isolates  of  endophytic  fungi  had  the  anti-microbial  activity  against  Staphylococcus  aureus, Escherichia coli, Bacillus subtilis and Salmonella typhi but only 3 endophytic fungi had the activity towards C. albicans.

ISOLASI MIKROBA ENDOFIT RANTING TUMBUHAN TRENGGULI (Cassia fistula L.) DAN AKTIVITAS ENZIM XILANASE

Jurnal Bahan Alam Indonesia Vol 6, No 4 (2008)
Publisher : Indonesian Research Gateway

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Isolation of endophytic microbe and the activity of xylanase enzym from the stems of Cassia fistula L has benn carried out. Fermentation of the microbe using xylan fermentation medium. The method used for the isolation was direct from the medium and then followed by macroscopic and microscopic identification. Evaluation of the enzym  xylanase activity was done using colorimeter at 521 nm wave length. The result showed 13 bacteria isolates and 18 fungi isolates. Three of bacteria isolates having activity to produce xylanase with the range 1,6685 U/ml (highest) and 0,1390 U/ml (lowest), while the activity of fungi endophyte were 41,2143 U/ml (highest) and 1.6652 U/ml (lowest).

AKTIVITAS ANTI BAKTERI EKSTRAK DAUN ILER (Coleus atropurpureus Benth) TERHADAP BEBERAPA BAKTERI GRAM (+) DAN BAKTERI GRAM (-)

Jurnal Bahan Alam Indonesia Vol 7, No 1 (2009)
Publisher : Indonesian Research Gateway

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Preliminary study of antibacterial activity of ethanolic extract of Iler leaves (Coleus atropurpureus Benth) has been carried out. Ethanolic extract were produced by maceration method and the evaluation of the antibacterial activity using diffusion method towards  Gram positive (Staphylococcus aureus, Bacillus subtilis) and Gram negative bacteria (Escherichia coli, Salmonella paratyphosa). The results showed that ethanolic extract had antibacterial activity of 1%, 10% and 20% against S.aureus and E.coli while the activity on B.subtilis and S. paratyphosa occurred at concentration of 10% and 20%. The statistic results showed significant differences between concentration and diameter inhibition zone of each bacterium.

PENAPISAN EFEK SITOTOKSIK, AKTIVITAS ANTIOKSIDAN DAN AKTIVITAS ANTIMIKROBA EKSTRAK DAUN MENGKUDU (Morinda citrifolia L)

Jurnal Bahan Alam Indonesia Vol 7, No 3 (2010)
Publisher : Indonesian Research Gateway

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Morinda citrifolia L, known as "mengkudu" has been used as traditional medicine. The purpose of this research was to explore the cytotoxic effect, antioxidant activity and antimicrobial activity of n-hexane, ethyl acetate and methanol extract from Morinda citrifolia L leaves. The cytotoxic effect was determined using MTT method, the antioxidant activity was evaluated using DPPH (2,2-diphenyl-1-picrylhidrazyl) method and agar diffusion method was used to determine the antimicrobial activity against Gram positive and negative bacteria. The result showed the IC50 value of ethyl acetate extract was 13.053 μg/ml. Antioxidant activity of the three extracts was low compared to vitamin C. The MIC of n-hexane extract showed the lowest MIC against Gram positive bacteria whereas ethyl acetate extract gave lowest MIC against Gram negative bacteria. The ethyl acetate extract could be further studied and developed based on its cytotoxic and Gram negative antimicrobial activity, whilst n-hexane extract on its potency as Gram positive antimicrobial agent.

AKTIVITAS ANTIMIKROBA EKSTRAK N-BUTANOL METABOLIT SEKUNDER KAPANG ENDOFIT 1.3.11 BUAH MAKASAR (Brucea javanica L.Mer) DENGAN VARIASI pH MEDIA FERMENTASI

Jurnal Bahan Alam Indonesia Vol 7, No 5 (2011)
Publisher : Indonesian Research Gateway

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Earlier study showed that the supernatant of fermentation process of 1.3.11 endophytic fungus has antimicrobial activity. This advanced research was condusted by varying the pH of fermentation medium which could affect the productivity of secondary metabolite. Antimicrobial activity of secondary metabolite of n-butanol extract was determined using disk method. The result showed that each pH of fermentation medium has antimicrobial activity to both positive and negative Gram bacteria and also to Candida albicans, but there was none of the pH has antimicrobial activity against Salmonella typhii. The acid fermentation medium has the best antimicrobial activity against Candida albicans.