Diameter Of Clear Zone Research Articles

Antibacterial Activity and Structure Elucidation of Salicin from Stem Bark of Salix tetrasperma ROXB

Salix tetrasperma Roxb. (Family Salicacaeae) is a plant that used as traditional medicine for anti-inflammatory, analgesic, reduces fever, and itching medicine. In this study was carried out extraction, isolation, structure elucidation of salicin from Salix tetrasperma Roxb. stem bark and it’s antibacterial activity. The extraction method was used the maceration method by n- hexane, ethyl acetate, and methanol solvents. Isolation of compound from ethyl acetate extract of Salix tetrasperma Roxb. stem bark using chromatography methods and obtained white solid (15 mg). The structure was elucidated using spectroscopic analysis, including Ultraviolet (UV), Infrared (IR), Nuclear Magnetic Resonance (NMR) and comparative literature, identified as salicin compound with molecule formula C 13 H 18 O 7 . Antibacterial activity was evaluated against Escherichia coli and Staphylococcus aureus bacteria using disk diffusion method. This compound has a great an antibacterial activity against Staphylococcus aureus bacteria with clear zone diameter of 10.2 ± 0.3 mm. This shows that the Salix tetrasperma Roxb. stem bark has great potential as a source of antibacterial compound

Uji Aktivitas Antibakteri Ekstrak Kasar Bawang Putih (Allium sativum ) terhadap Bakteri Shigella dysenteriae yang Resisten terhadap Ampisilin

ABSTRACT Background : Gastrointestinal infections, especially caused by Shigella dysenteriae bacteria is still a health issue in Indonesia. The number of ampicillin resistance to S. dysenteriae has been widely reported. Garlic (Allium sativum) is a traditional medicine that efficacy as an antibacterial. Garlic contains an organosulfur compound Allicin. Research Purpose: This study aimed to determine the antibacterial activity of garlic rule extract on the growth of S. dysenteriae bacteria that are resistant to ampicillin. Research Methods: This study used the experimental with post test only control group design. The antibacterial activity test was determined based on the diameter of clear zone formed. Test were performed on garlic filtrate, ciprofloxacin as a positive control and aquades as a negative control. Each treatment was performed three times. Research Result: This research showed that rule extract of garlic (A. sativum) had a clear zone against S. dysenteriae that was resistant to ampicillin. Conclusion: The conclusion of this research was that rule extract of garlic (A. sativum) had an antibacterial activity against S. dysenteriae which resistant to ampicillin. Keywords : Antibacterial activity, Garlic (A. sativum), S. dysenteriae

POTENTIAL OF CELLULASE OF PENICILLIUM VERMICULATUM FOR PREPARATION AND CHARACTERIZATION OF MICROCRYSTALLINE CELLULOSE PRODUCED FROM α-CELLULOSE OF KAPOK PERICARPIUM (CEIBA PENTANDRA)

Objective: This study aimed to find psychochemical properties of microcrystalline cellulose (MCC) obtained from α-cellulose kapok pericarpium. Methods: The cellulase activity was screened by clear zone and sugar reduction method. The enzym from selected mold was purified by diethylaminoethyl (DEAE) chromatography. α-cellulose of kapok pericarpium was hydrolyzed using the purified cellulase enzymes. Microcrystalline cellulose (MCC) identified by Fourier transform infrared (FTIR) spectrometry, and qualitative analysis test. The samples were characterized for pH test, x-ray diffraction (XRD), and particle size analyzer (PSA). Results: The optimum cellulase activity was shown by Penicillium vermiculatum. It’s clear zone diameter around 3 cm and the cellulase activity was 67.73±0.25 mU/ml. The strongest cellulase activity was detected from 1st fraction (P1) out of 6 column fractions with optimum activity at 1.177±2 mU/ml. The optimal conditions for microcrystalline cellulose (MCC) preparation were at 50 ˚C, for 2 ours, using 20 ml of acetate buffer pH 5 and 2 ml of cellulase enzyme. Microcrystalline cellulose (MCC) obtained at 78% w/w and its FTIR spectrum and x-ray diffractogram similar to reference while the pH of MCC was fulfilled requirements of The United States Pharmacopoeia 2007. Conclusion: The use of purified enzyme of cellulase has succeded in microcrystalline cellulose (MCC) preparation andmicrocrystalline cellulose (MCC) obtained was 78% w/w, with similar characteristics to reference (Avicel PH 101) and the pH of MCC was fulfilled requirements of The United States Pharmacopoeia 2007.

Isolation and identification of low density polythene-degrading bacteria from soil of North West of Algeria

Plas c bags (Low Density Polyethylene (LDPE) belong to the polymers, which plays a very important role in our daily lives by their diversi ed applica on. However, the accumula on of the plas c bags in the environment cons - tutes a serious problem and a real source for visual nuisance, pollu on of soil and marine environments. Furthermore, their biodegradation was the safest method of breakdown that possibly leaves behind less toxic residues and showed poten al of bio-geo chemical cycling of the substrate. The aim of the present work was the characterization of the isolated bacterial strains from a municipal land ll area of Tlemcen, North West Algeria, which were implicated by the biodegrada on ability of the Low Density Polyethylene. The degradation of the Low Density Polyethylene was inves gated by studying the bacterial growth of the isolated, inoculated on a solid culture medium, which was composed of LDPE as the sole carbon source with and with- out a nitrogen source and the selec on was based by the determination of the produced diameter of hydrolysis clear zone on the surface. Furthermore, the isolated, selected degrading Low Density Polyethylene bacterial ML002 has been iden ed by the study of their morphological, biochemical charac- teris cs and the ampli ca on of the fragment, coding the region of ARN 16S. The use of the API system indicated their belonging to the genus Bacillus Cereus, which has reduced the weight of LDPE by 0.26, 1.28, 1.53% a er 30, 90, 120 days respec vely. Furthermore, the amplified of the fragment, coding the region of ARN 16S by the isolated, selected bacterial ML002 indicated a similarity of 99.394% with Bacillus wiedmannii and Bacillus proteolyticus and 99.293% homology with Bacillus toyonensis, Bacillus cereus and Bacillus thuringiensis.

Biofertilizer/biopesticide potentiality of zinc solubilizing Pseudomonas aeruginosa FA-9 and Enterobacter sp. FA-11 isolated from the wheat rhizosphere grown in arid zone

Bacterial strains were obtained from wheat rhizosphere and screened for zinc solubilization on agar plates. Two strains FA-9 and FA-11 were found efficient for Zn solubilizing activity and were identified as Pseudomonas aeruginosa and Enterobacter sp. by 16S rRNA and gyrase (gyrB) genes analysis, respectively. The strain FA-9 produced a clear zone diameter of 63 mm, 60 mm and 51 mm on agar plate amended with different zinc ores. The strain FA-11 produced a zone diameter of 17 mm with zinc carbonate and 20 mm with zinc oxide while no zone was observed with zinc phosphate. Both strains showed no visible activity with ZnS ore. Similarly, FA-9 and FA-11 increased maximum soluble zinc content (102 µg ml−1 and 45 µg ml−1) from zinc carbonate ore as compared to zinc oxide ore (102 µg ml−1 and 45 µg ml−1) in liquid broth. It was noted that both strains exhibited less potential (7 µg ml−1 and 0.57 µg ml−1) to solubilize ZnS ore in liquid broth. A comparison between agar plate assay and liquid broth quantification shows that agar plate assay does not present the solubilizing potential of ZSB precisely. The strains FA-9 and FA-11 produced auxin with l-tryptophan (3.25 µg ml−1 and 2.86 µg ml−1) and without l-tryptophan (1.23 µg ml−1 and 1.02 µg ml−1). Both strains expressed exo-polysaccharides (EPS) and siderophores activity along with phosphate (P) solubilization, ACC deaminase and antifungal activities. The ACC deaminase and N-fixation activity was confirmed by the amplification of acdS and nifH genes respectively.

Isolation of Cellulose Degradation Bacteria (CDB) from acid soil as a potential candidate of organic waste degradation

Background: The study aimed to obtain CDB with high degraded activities, determined growth curve, protein content, and cellulase maximum activity (exoglucanase and endoglucanase). Method: The cellulose activity calculated according to Miller (1959), protein content was measured by Bradford method with bovine serum albumin (BSA) as a standardize protein. Result: Six isolates of CDB were found as potential degradation of organic waste (Km25, Sr75, Jm, U6, G8, and Km13). Growth curve, protein level, and protein maximum activity occurred on day-3. The largest diameter of clear zone of six isolates was Km25, Sr75, Jm, U6, G8, and Km13 (3.32, 3.31, 2.41, 2.36, 2.19, and 2.04 mm, respectively). Endoglucanase and exoglucanase maximum activity were 0.011-0.402 IU/mL and 0.0028-0.155 IU/mL, respectively. Conclusion: Six isolates showed high activities of cellulase with diameter of clear zone ≥ 2 cm (Km25, Sr75, Jm, U6, G8, and Km13). Growth maximum curve was on day-3. Highest endo- and exoglucanase activities were on day-3 (0.402 IU/mL and 0.155 IU/mL, respectively) in Jm isolate.
 Keywords: Identification, degradation, clear zones, cellulase.

Screening and Molecular Characterization of Cellulase Producing Actinobacteria from Litchi Orchard

Background:Cellulolysis is of considerable economic importance in laundry detergents, textile and pulp and paper industries and in fermentation of biomass into biofuels.Objective:The aim was to screen cellulase producing actinobacteria from the fruit orchard because of its requirement in several chemical reactions.Methods:Strains of actinobacteria were isolated on Sabouraud’s agar medium. Similarities in cultural and biochemical characterization by growing the strains on ISP medium and dissimilarities among them perpetuated to recognise nine groups of actinobacteria. Cellulase activity was measured by the diameter of clear zone around colonies on CMC agar and the amount of reducing sugar liberated from carboxymethyl cellulose in the supernatant of the CMC broth. Further, 16S rRNA gene sequencing and molecular characterization were placed before NCBI for obtaining recognition with accession numbers.Results:Prominent clear zones on spraying Congo Red were found around the cultures of strains of three groups SK703, SK706, SK708 on CMC agar plates. The enzyme assay for carboxymethylcellulase displayed extra cellulase activity in broth: 0.14, 0.82 and 0.66 µmol mL-1 min-1, respectively at optimum conditions of 35°C, pH 7.3 and 96 h of incubation. However, the specific cellulase activities per 1 mg of protein did not differ that way. It was 1.55, 1.71 and 1.83 μmol mL-1 min-1. The growing mycelia possessed short compact chains of 10-20 conidia on aerial branches. These morphological and biochemical characteristics, followed by their verification by Bergey’s Manual, categorically allowed the strains to be placed under actinobacteria. Further, 16S rRNA gene sequencing, molecular characterization and their evolutionary relationship through phylogenetics also confirmed the putative cellulase producing isolates of SK706 and SK708 subgroups to be the strains of Streptomyces. These strains on getting NCBI recognition were christened as Streptomyces glaucescens strain SK91L (KF527284) and Streptomyces rochei strain SK78L (KF515951), respectively.Conclusion:Conclusive evidence on the basis of different parameters established the presence of cellulase producing actinobacteria in the litchi orchard which can convert cellulose into fermentable sugar.

Antibacterial Activity Test of Indigenous Yeast from Sapodilla Fruit against Staphylococcus aureus and Escherichia coli

The research aimed to identify indigenous yeast antibacterial activity from sapodilla fruit against Escherichia coli and Staphylococcus aureus, which conducted by experimental methods and followed by descriptive analysis. This study was done by the isolation of indigenous yeast, macroscopic and microscopic identification, yeast identification using RapID Yeast Plus System, antibacterial test by measuring the clear zone diameter, testing of pathogenic bacteria viability against indigenous yeast and identification of organic acid produced by yeast. The results of yeast isolation obtained 1 isolate (Saccharomyces cereviseae 1) from fruit and 3 isolates form sapodilla skin (S.cereviseae 2, Candida famata, and Pichia anomala) which had antibacterial activity against E. coli and S. aureus except C. famata isolates. Isolates with the largest antibacterial activity against E. coli and S. aureus based on the clear zone diameter were S. cerevisiae (2) isolates. The results of organic acid analysis by HPLC found that S.cerevisiae (2) isolate produced the highest organic acid namely acetic acid as much as 2.442 mg mL -1. Key words : antibacterial, organic acid, sapodilla fruit, yeast

Isolation of <i>Pseudomonas</i> with the Ability to Solublize Phosphate from Inorganic and Organic Phosphate Sources

A total of eleven phosphate solubilizing bacteria were isolated from rhizosphere soil of tomato (Solanum lycopersicum) plants. Isolates designated as PSB1, PSB2, PSB4, PSB5 and PSB7 were selected out of eleven isolates based on their clear zone diameters. They were characterized to generic level and found to belong to Pseudomonas spp. Tricalcium phosphate (TCP), rock phosphate (RP) and bone phosphate (BP) solubilizing efficiency of these isolates was studied in the laboratory at 5, 10, and 15 days of incubation. All bacterial isolates solubilized significantly (P< 0.05) higher amounts of TCP over uninoculated control. The highest P was release upon 5 days of incubation was recorded from the isolate PSB1 with 7.64 mg/50ml in the medium and the least P release was recorded by isolates PSB4 and PSB5 with a P content of 4.79 mg /50ml. At 10th day of incubation, the highest P release was recorded by isolates PSB2 (8.19 mg/50ml), PSB1 (8.10mg/ 50ml). At 15th days of incubation, the highest P release was recorded by isolate PSB7 (11.77 mg/ 50ml) and PSB2 (11.33 mg/ 50ml) and the least P released was recorded by isolate PSB5 (5.44 mg/ 50ml)). Under RP solubilization, the p released from all isolates upon 5 days of incubation was not significant as compared to the control. At 10th day of incubation, the highest P release was recorded by isolates PSB5 (5.56 mg/50ml), PSB7 (5.02 mg/ 50ml) and the least P release was observed by isolate PSB4 (3.46mg/g). At 15th days of incubation, the highest P release was recorded by isolate PSB7 (7.928 mg/ 50ml) and the least P release was recorded by isolate PSB4 (4.025mg/ 50ml). Regarding BP, the p released from all isolates except isolate PSB7 (3.020mg /50ml) upon 5 days of incubation was not significant as compared to the control. At 10th day of incubation, the highest P release was recorded by isolates PSB5 (7.37 mg/50ml), PSB7 (7.025 mg/ 50ml) and the least P released was observed by isolate PSB2 (5.47 mg/50ml). At 20th days of incubation, the highest P release was recorded by isolate PSB7 (11.09 mg/ 50ml) and the least P release was recorded by isolate PSB4 (9.06 mg/ 50ml). So, as conclusion all Psedomonas isolates were able to release P from both inorganic and organic phosphate sources.

FORMULASI DAN UJI EFEKTIVITAS ANTIBAKTERI EDIBLE FILM EKSTRAK BIJI PINANG (Areca catechu Linn)

ABSTRACTThe tannins contained in areca nut extract can inhibit the growth of Streptococcus mutans bacteria. To facilitate the use of betel seed extract as an antibacterial, it was formulated into an edible film with a concentration of 2.5%, 3% and 3.5%. Making edible films using solvent casting method and testing antibacterial activity using blood agar diffusion method. The edible film of areca nut extract made an evaluation included organoleptic, thickness, brittleness, water content, pH, and swelling test. The evaluation results of betel nut edible fim extract showed almost the same characteristics as the edible fim that had been circulating in the market which was used as a comparison. The presence of antibacterial activity from edible pinang seed extract is marked by the formation of clear zones. The test results of antibacterial edible activity of areca nut extract films 2.5%, 3% and 3.5% showed the average diameter of clear zone was 14.96 mm, 15.49 mm and 17.05 mm which were included in the strong category.Keywords: Areca catechu Linn, antibacterial, edible film, tannin, streptococcus

PENGARUH PEMBENTUK FILM PVA PADA SEDIAAN MASKER GEL PEEL-OFF EKSTRAK ETANOL BAWANG DAYAK (Eleutherina palmifolia (L.) Merr) TERHADAP AKTIVITAS ANTIBAKTERI Staphylococcus aureus PENYEBAB JERAWAT

ABSTRACTDayak Onion (Eleutherine palmifolia) is one of the plants that can inhibit Staphylococcus aureus bacteria that causes acne. This study aims to determine the effect of polyvinyl alcohol on peel-off gel mask preparations from onion dayak ethanol extract on the antibacterial activity of Staphylococcus aureus that causes acne. Extraction using maceration method with 96% ethanol. The concentration of dayak onion extract was made at 5%, 7.5%, and 10% and the peel-off gel mask formula from dayak onion extract with variations in extract concentration were 5%, 7.5%, and 10% based on Polyvinyl Alcohol (PVA). Antibacterial activity is carried out using the disk disc method. In testing the inhibition of dayak onion extract at a concentration of 5% resulted in a clear zone of 15.5 mm, a concentration of 7.5% and 10% producing the same clear zone diameter of 17.1 mm. The peel-off gel mask in formulation I obtained a clear zone diameter of 8.7 mm, formulation II the diameter of the clear zone was 12.2 mm, and formula III the diameter of the clear zone was 15.3 mm. The results of this study indicate that on dayak onion extract has a greater inhibitory power compared to the dayak onion extract peel-off gel mask preparation. It can be concluded that the PVA in the preparation affects the amount of inhibitory power on the bacteria forming acne, namely Staphylococcus aureus.Keywords: Dayak onion extract, Staphylococcus aureus, disk discs, antibacterial inhibitory test, acne-causing bacteria, peel-off gel mask.

Anti-thrombosis Activity of Drone Apis mellifera Pupae Extracts

Drones of honeybee (Apis mellifera) have been regarded as a useful value only when mating with queen bee. However, the drone pupae have been reported to be nutritionally valuable, and a potential beekeeping product. In this study, drone pupae extracted with 5% acetic acid were used to measure anti-thrombosis related fibrinolytic activity using Strup and Mullertz fibrin plate method. As a result, the drone pupae extract showed higher effect of fibrinolytic activity(clear zone diameter 20.83mm) compared to the human plasmin (clear zone diameter 12.93mm) used as a positive control. It was suggested that the extract of drone pupae can be developed as a functional material helping prevention or treatment of various vascular diseases.

Use of Orange Oil Loaded Pectin Films as Antibacterial Material for Food Packaging.

This study aims to develop orange oil loaded in thin mango peel pectin films and evaluate their antibacterial activity against Staphylococcus aureus. The mango peel pectin was obtained from the extraction of ripe Nam Dokmai mango peel by the microwave-assisted method. The thin films were formulated using commercial low methoxy pectin (P) and mango pectin (M) at a ratio of 1:2 with and without glycerol as a plasticizer. Orange oil was loaded into the films at 3% w/w. The orange oil film containing P and M at ratio of 1:2 with 40% w/w of glycerol (P1M2GO) showed the highest percent elongation (12.93 ± 0.89%) and the lowest Young’s modulus values (35.24 ± 3.43 MPa). For limonene loading content, it was found that the amount of limonene after the film drying step was directly related to the final physical structure of the film. Among the various tested films, P1M2GO film had the lowest limonene loading content (59.25 ± 2.09%), which may be because of the presence of numerous micropores in the P1M2GO film’s matrix. The inhibitory effect against the growth of S. aureus was compared in normalized value of clear zone diameter using the normalization value of limonene content in each film. The P1M2GO film showed the highest inhibitory effect against S. aureus with the normalized clear zone of 11.75 mm but no statistically significant difference. This study indicated that the orange oil loaded in mango peel pectin film can be a valuable candidate as antibacterial material for food packaging.

FORMULASI DAN AKTIVITAS ANTIBAKTERI SEDIAAN GEL HAND SANITIZER DARI EKSTRAK ETANOL BIJI MANGGA HARUM MANIS ( Mangifera indica L.) TERHADAP Escherichia coli DAN Staphylococcus aureus

Mangifera indica L. is a plant that has antibacterial activity. One of the plant‟s parts that has antibacterial activity on mango is the seed. Natural antibacterial content in the form of tannins contained in ethanol extract of Harum Manis mango seed is considered being effective towards Escherichia coli and Staphylococcus aureus bacteria, which are the main cause bacteria in diarrhea and other skin infections. The objective of this study is to obtain a good and effective formulation to remove Escherichia coli and Staphylococcus aureusbacteria in order to produce natural hand- made antiseptic gel preparations. The diameter of clear zone produced by gel preparation of hand sanitizer ethanol extract of mango seed on Escherichia coli bacteria with gel concentration of 2; 2.5 and 5% respectively - 10.86, 1.74, and 13.15 mm, while on a positive control, it is 7.47 mm. On the other hand, the diameter of clear zone in Staphylococcus aureus with gel concentration of 2; 2.5 and 5% respectively - 10.14, 10.58, and 12.83 mm, while on the positive control, it is 7.43 mm. Based on Anovaone way test followed by Duncan test show a significant difference (significant effect) between the concentration. It can be concluded that the preparation of 70% ethanol extract gel having antibacterial effectiveness towards Escherichia coli and Staphylococcus aureus and effective gel preparation evaluation results are at 2,5% concentration

Screening and identification of biosurfactant producing marine bacteria from the Caspian Sea

Marine microorganisms have unique physiological properties and novel metabolites such as biosurfactant to live in extreme habitats (e.g. oil polluted environment). The aim of this study was to characterize some biosurfactant-producing bacteria collected from sediment and seawater samples from the Caspian Sea. These bacteria were isolated using enrichment method in ONR7a medium with crude oil as sole carbon source. Five screening tests were used for selection of the bacteria including hemolysis in blood agar, oil spreading, drop collapse, emulsification activity and bacterial adhesion to hydrocarbon test (BATH). The bacteria isolated were identified using molecular methods. Eighteen biosurfactant-producing bacteria (from 44 different colonies isolated) were selected and 9 isolates were screened as predominant biosurfactant producer belonging to Pseudomonas stutzeri (Strain S1-4-2), P. balearica (Strain S1-4-1), Pseudomonas sp. (Strain S2-1), P. stutzeri (Strain R3-2), P. chloritidismutans (Strain K4-1), Achromobacter xylosoxidans (Strain K4-3), Acinetobacter radioresistens (Strain S1-2), Acinetobacter calcoaceticus (Strain K4-2) and Citrobacter freundii (Strain R3-1). The largest clear zone diameters for oil spreading method observed for P. balearica strain S1-4-1 (14 mm). Also, this strain has the best emulsification activity (100 %); surface hydrophobicity (BATH: 77 %) and reduction of surface tension (34.5 mN m-1). Thus, this strain selected as the best isolated strains. The results of this study confirmed that there is sufficient diversity of biosurfactant-producing bacteria in the Caspian Sea and the environmental problem can be resolved by application of these bacteria in petrochemical waste water.

Antifungal activity of some toothpastes and antiseptic solutions against yeasts isolated from the oral cavity of cancer radiated patients

This study aimed to investigate the antifungal efficacy of different types of toothpaste andsome antiseptic solutions with varying concentrations against yeast isolated from the oral cavityof radiated patients suffering from head and neck cancer in radiotherapy unit in Suez CanalUniversity teaching’s hospital, Egypt. Eight different species of yeast were isolatedtaxonomically, assigned to 1 phylum, two classes, one order, and three families and identifiedphenotypically. The antifungal activity of five kinds of toothpaste and three antiseptic solutionswere investigated by agar well diffusion method against the eight isolated yeasts namely:Candida glabrata, C. albicans, C. tropicalis, C. kefyr, C. guilliermondii, C. parapsilosis,Dipodascus geotrichum and Geotrichum beigelii and expressed as the value of the clear zonediameter, however, Their activity was expressed by minimum inhibitory concentration (MIC).Results showed that toothpaste no 1, 3, 4 and 5 were effective against all tested taxa by variableinhibitory degrees. However, no. 2 showed efficacy only against C. glabrata, C. guilliermondii,and C. parapsilosis, Antiseptic solution no 1 showed the highest efficacy against all testedspecies except C. kefyr while no. 2 was effective against all the tested species except two taxaand finally no. 3 exhibited good antifungal activity against all the tested taxa. In conclusion, werecommended the use of toothpaste contained sodium monofluorophosphate, or sodium fluoride,as an active component followed by a mouthwash which contained Chlorohexidine orCetylpridinium or miswak as active components to prevent colonization and development of oralcandidiasis, especially in cancer patients.

UJI DAYA HAMBAT EKSTRAK Chaetoceros calcitrans TERHADAP BAKTERI Aeromonas salmonicida

The impact of use antibiotic can cause problems on aquaculture such as bacterial resistance and residues for consumer. Therefore, one of alternatives that can be used to inhibit or kill bacteria, one of which is the use of plankton Chaetoceros calcitrans . This research aims to know the influence of the granting C. calcitrans extract that can inhibit or kill bacteria and know the best dose for inhibit A. salmonicida bacteria. The experiment method used randomize completely design consist of four treatment and two control. A treatment (0.3 ppm), B treatment (15.3 ppm), C treatment (30.3 ppm), D treatment (45.3 ppm), positive control (add chlorampenicol 5ppm) and negative control (without extract). The result of research showed the highest average of clear zone diameter in D treatment 5.37 mm and the lowest average of clear zone diameter in A treatment 2.89 mm, with linear equation y = 3.2067 + 0.0509x and coefficients R 2 = 0.9262. The relationship between C. calcitrans extract in inhibitng gwoth of A. salmonicida bacteria showed that the average diameter of the clear zone has increase in line with the addition of extracts. Form the research showed that the extract of C. calcitrans was bacteriostatic (inhibits the growth of bacteria) and best obtained at the treatment dosage of D with a dose of 45.3 ppm and included into the category of middle.

Strain Screening from Traditional Fermented Soybean Foods and Induction of Nattokinase Production in Bacillus subtilis MX-6.

The plasminogen-free fibrin plate assay method was used to isolate Bacillus subtilis MX-6, a strain with high production of nattokinase from Chinese douchi. The presence of aprN, a gene-encoding nattokinase, was verified with PCR method. The predicted amino acid sequence was aligned with homologous sequences, and a phylogenetic tree was constructed. Nattokinase was sublimated with ammonium sulfate, using a DEAE-Sepharose Fast Flow column, a CM-Sepharose Fast Flow column and a Sephadex G-75 gel filtration column. SDS-PAGE analysis indicated that the molecular weight of the purified nattokinase from Bacillus subtilis MX-6 was about 28kDa. Fermentation of Bacillus subtilis MX-6 nattokinase showed that nattokinase production was maximized after 72h; the diameter of clear zone reached 21.60mm on the plasminogen-free fibrin plate. Nattokinase production by Bacillus subtilis MX-6 increased significantly after supplementation with supernatant I, supernatant II and soy peptone but decreased substantially after the addition of amino acids. This result indicated that the nattokinase production by B. subtilis MX-6 might be induced by soybean polypeptides. The addition of MgSO4 and CaCl2 increased B. subtilis MX-6 nattokinase production.

Isolate Actinomycetes SA32 Origin of Segara Anakan Mangrove Rhizosphere and its Capability in Inhibiting Multi-Drugs Resistant Bacteria Growth

The screening of antibiotic-producing actinomycetes from the mangrove environment is increasing to search for bioactive compounds capable of inhibiting the growth of Multi-Drugs Resistant (MDR) bacteria. The research aimed to isolate actinomycetes from mangrove mud rhizosphere in Segara Anakan lagoon that is capable of producing bioactive compounds inhibiting MDR bacterial growth, to characterize phenotypically along with 16S rRNA gene sequence, to cover out inhibition potencies toward MDR bacterial growth, and to characterize antibacterial compounds produced. Isolate actinomycetes SA32 was isolated from rhizosphere mud of Rhizophora mucronata from east Segara Anakan. It showed fragmented aerial mycelium similar to Streptomyces sp. NEAE-102; however, 16S rRNA gene sequence analysis yielded 96% similarity to Streptomyces sp. N56. In antagonism assay, it inhibited the growth of MDR bacteria Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas areuginosa, Enterococcus cloacae and Enterobacter sp. The diffusion antagonism assay results in the highest antibacterial activity against S. aureus with clear zone diameter 20 mm. The result of Minimum Inhibitory Concentration (MIC) assay showed that 20% crude extract has been able to inhibit the growth of MDR bacteria characterized by the decrease of turbidity. The antibacterial compound produced was not known yet but has an Rf value of 0.7-0.9 in the TLC test. Isolate actinomycetes SA32 is potent to be developed as an MDR antibacterial substance source and it is proposed as a new strain of Streptomyces.

Screening of secondary metabolite from waterfall and marine bacteria as biocontrol agent

Biocontrol agent often used to reduce sources of disease, such as bacteria, fungi, and mosquito. Emergence of antibiotic-resistant bacteria and fungi are increasing due to excessive used of antimicrobial substances. Chemical substances used to kill mosquito larvae in water environment might also be dangerous for human and environment. Hence, a biocontrol agent is required to be safe and effective. The purpose of this study is to discover bacteria from waterfall and marine water which has a good potential as biocontrol agent. Four waterfall bacteria (S 1.1, S 2.1, S 3.1, and S 3.2) and three marine bacteria (A 1.1, A 1.2, and PA 1.3C) were proved to have antifungal activity towards pathogenic fungal. PA 1.3A was the only marine bacteria which proved to have antimicrobial activity towards Escherichia coli and Streptococcus agalactiae. Extraction of antimicrobial substance using several solvents resulted in larger clear zone diameter and inhibition of other bacteria, such as Bacillus licheniformis and Staphylococcus aureus. Extracted antimicrobial substance of PA 1.3A was also proved to be non toxic towards Aedes aegypti larvae through larvicidal assay and mammals through brine shrimp lethality assay (LC50 > 1000μg/mL).