Incubator Shaker Research Articles

In silico, in vitro and in vivo metabolite identification of brexpiprazole using ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.

Brexpiprazole is a novel serotonin-dopamine activity modulator approved by the USFDA in July 2015 for the treatment of schizophrenia and as an adjunctive therapy with other antidepressants for major depressive disorder in adults. However, limited numbers of metabolites are reported in the literature for brexpiprazole. Our prime intent behind this study is to revisit metabolite profiling of brexpiprazole and to identify and characterize all possible in vitro and in vivo metabolites. Firstly, the site of metabolism for brexpiprazole was predicted by a Xenosite web predictor model. Secondly, in vitro metabolite profiling was performed by incubating the drug individually with rat liver microsomes, human liver microsomes and rat S9 fraction at 37°C for 1 h in incubator shaker. Finally, for in vivo metabolite identification, a 50 mg kg-1 dose of brexpiprazole was administered to male Sprague-Dawley rats and the presence of various metabolites was confirmed in rat plasma, urine and feces. The predicted atomic site of metabolism was obtained as a color gradient by the Xenosite web predictor tool and, from this study, probable metabolites were listed. In total, 14 phase I and 2 phase II metabolites were identified and characterized in the in vitro and in vivo matrices using ultra-high-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry (UHPLC/QTOF-MS/MS). The majority of metabolites were found in the sample incubated with human liver microsomes and in rat urine, while in the other matrices only a few metabolites were detected. All the 16 metabolites were identified and characterized using UHPLC/QTOF-MS/MS. The study revealed that brexpiprazole is metabolized via hydroxylation, glucuronidation, S-oxidation, N-oxidation, dioxidation, oxidative deamination, N-dealkylation, etc.

Features of low-temperature storage of streptomyces strains - producers of glycosidaz inhibitors

The aim of the work is to study the inhibitory activity of the Streptomyces lucensis VKPM Ac-1743 and Streptomyces violaceus VKPM Ac-1734 strains stored at –18 °C in a 15 % glycerol solution and in a 0.9 % sodium chloride solution.The object of the study was the actinomycete strains of S. lucensis VKPM Ac-1743 and S. violaceus VKPM Ac-1734 producers of glycosidase inhibitor, which is a micro-ingredient for creating products with a low glycemic index. The storage time was nine months. Bookmark storage was performed by flushing with a taped agar starchcontaining environment of Capek. For comparison, we studied the properties of cultures stored at +4 °C without cryoprotectant.Deep cultivation of Streptomyces strains was carried out in a periodic manner on a medium containing corn starch hydrolyzate under the Multitron incubator shaker (INFORS,Switzerland).Inhibitory activity was determined in inactivated native solutions by a colorimetric method with respect to pancreatic α-amylase, a test glycosidase involved in carbohydrate metabolism, and whose activity was chosen as a criterion for assessing the hypoglycemic action of the inhibitor. The proteinase activity of the inactivated native solution was determined by a modified method using a casein substrate.The experimental data were processed using the methods of mathematical statistics and Excel XP programs.As a result of the research, it was established that the actinomycete strains of S. lucensis and S. violaceus, the producers of glycosidase inhibitors, can maintain inhibitory activity during low-temperature storage for nine months.For the S. lucensis actinomycete strain, stored in a 15 % glycerol solution at – 18 °C, the maximum inhibitory activity is 48 hours in the biotechnological process and is (3686 ± 300) IU/cm3 native solution, and for theviolaceus strain – (3150 ± 200) IU/cm³of the native solution, respectively.For the strain of actinomycete S. lucensis, stored in 0.9 % sodium chloride solution at a temperature of – 18 °C, the maximum inhibitory activity accounts for 72 h of the biotechnological process and is (2600 ± 200) IU/cm³ of the native solution, and for the strain S. violaceus the maximum inhibitory activity accounts for 24 hours of the biotechnological process and is (3530 ± 200) IU/cm³ of the native solution.At a storage temperature of +4 °C, the inhibitory activity for the strain of actinomycete S. lucensis is (560 ± 20) IU/cm³of native solution, and for the strain of S. violaceus – (1747 ± 100) IU/cm³ of native solution, respectively. On the basis of the data obtained, it can be concluded that a temperature of –18 °C is preferred for long-term storage.During the cultivation of Streptomyces strains, proteinase activity ranged from (0,012 ± 0,001) U/cm³ to (0,072 ± 0,002) U/cm³.The obtained data can be applied in further studies to develop conditions for long-term storage of collection crops.

Sustainable synthesis of pectinolytic enzymes from citrus and Musa acuminata peels for biochemical oxygen demand and grease removal by batch protocol

In recent years, oil and grease has been identified as an emerging pollutant of concern (EPC) in wastewater stream as it can disturb the ecology and wastewater treatment process efficiency. The highest contributor to oily wastewater among domestic wastewater is from kitchen greywater. One of the alternatives to address this problem is the application of enzyme. The production of enzyme by using organic waste has gained significant attention in the recent years due to sustainable demand from it. In this study, pectinolytic enzyme was produced through simplified fermentation from discarded citrus peels that possess high lipase content. Three batches of treatment which consist of the control sample (solely wastewater), 25% (v/v) citrus enzyme + wastewater and 50% (v/v) citrus enzyme + wastewater was incubated in an incubator shaker for 10 days at 30 °C and 150 rpm. The wastewater analysis was performed at a regular interval of 48 h. The parameters monitored were pH, BOD5 and oil and grease. Laboratory work has demonstrated that 25% (v/v) pectinolytic enzyme was able to remove BOD5 and oil and grease about 10% better than 50% (v/v) pectinolytic enzyme. The percentage of removal achieved by 25% (v/v) pectinolytic enzyme was 39.83 ± 9.50 mg/L and 64.21 ± 1.12 mg/L, respectively. However, it was observed that enzyme was less effective in removing BOD5 as the solution contains organic matter that increases the total organic matter in the wastewater mixture.

Synthesis and characterization of hydroxyapatite from bovine bone for production of dental implants.

This paper presents a study on the synthesis and characterization of hydroxyapatite that can be used to produce dental implants or serves as a replacement for other body hard tissues. The bioceramic material (hydroxyapatite) was derived from bovine bone samples after they were de-fatted and processed to produce particles measuring less than 250 μm. Processed bone powders were calcinated at 750℃ to obtain calcium phosphate compound (hydroxyapatite). This indicated complete elimination of the organic phase of the bone. The synthesized hydroxyapatite was physico-chemically characterized using Fourier transform infrared spectroscopy (FTIR), an optical microscope and scanning electron microscopy (SEM). Hydroxyapatite powder was sintered into a block and then characterized mechanically by measuring its hardness using a Rockwell hardness testing machine. The hydrophilicity and hydrophobicity of the sintered blocked specimens were studied. The average wettability of the substrate, which was a characteristic of the contact angle, was found to be approximately 31.73 degrees. This contact angle value is a measure of the material hydrophilicity. The biostability/corrosive resistance of the sintered samples was simulated in vitro in phosphate buffered saline solution (pH 7.4) by incubating in an optical incubator shaker (set at 60 revolutions per minute) to mimic physiological conditions. The results showed that hydroxyapatite can be extracted economically from a natural source such as bovine bone and can be employed as a restorative biomaterial for dental implants and hard tissue replacement.

Effects of substrate concentration on bioethanol production from oil palm empty fruit bunches with simultaneous saccharification and fermentation (SSF)

Oil palm empty fruit bunches (OPEFB) are industrial wastes generated from the processing of palm oil. OPEFB has the potential to be feedstock for bioethanol production due to its high cellulose content. In this paper, ethanol production is conducted with Simultaneous Saccharification and Fermentation (SSF) method, which combined hydrolysis (saccharification) and fermentation process simultaneously in one reactor to create a faster conversion time and minimize costs. The aim of the study was to determine the effect of substrate concentration on the growth of microorganisms and the yield of bioethanol produced through the SSF method. Pretreatment of OPEFB to decompose lignin is done by soaking it in alkaline solution. Hydrolysis is carried out enzymatically with cellulase enzyme within 48 hours in incubator shaker. The process is then followed by fermentation within 12 hours by adding Zymomonas mobilis inoculum. The research method was done experimentally with descriptive analysis, which included the effects of OPEFB substrate concentration (3%, 6%, 9%, 12%, 15%) on ethanol production. The result shows that the high concentrations of substrate in the medium increase the concentration of glucose, in which the highest concentration is 3.2 g/L. The higher the substrate loading resulted in lower hydrolysis yield and ethanol yield. The highest ethanol concentration is 0.25 g/L, obtained from fermenting 9% OPEFB loading in 3 hours.

Optimization of myco-synthesized silver nanoparticles by response surface methodology employing Box-Behnken design

The objective of current study was to optimize the average diameter size of biosynthesized Silver Nanoparticles (AgNPs) induced by Penicillium citrinum. The AgNPs were prepared with a spherical shape and good monodispersity, determined by scanning electron microscopy (SEM), atomic force microscopy (AFM) and photon correlation spectroscopy (PCS). Besides, response surface methodology (RSM) was conducted to optimize the average diameter size of AgNPs by employing Box-Behnken design (BBD). The effective factors in this process were concentration of AgNO3 (mM) (X1), pH of solution (X2), temperature of shaker incubator (°C) (X3), rate of shaker incubator (rpm) (X4) and time of incubation (hour) (X5). The R2 (0.8894) value of the obtained model indicated a good correlation between the observed and predicted values. Overall, it could be concluded that multivariate analysis facilitated to find out the optimum conditions for the biosynthesis of AgNPs induced by P. citrinum with the reduced time and cost.

Microwave Irradiation-Assisted Chitosan Hydrolysis Using Cellulase Enzyme

The influence of microwave irradiation on the chitosan hydrolysis catalyzed by cellulase enzyme was studied. The hydrolyzed chitosan was characterized by measuring its viscosity and reducing sugar. Further, it was also characterized by Fourier-Transform Infrared Spectroscopy (FT-IR), X-ray Diffraction (XRD), and Scanning Electron Microscope (SEM). The classical Michaelis-Menten kinetic parameters were measured by analyzing the amount of reducing sugars. The results were compared with the hydrolysis by using conventional shaker incubator. The hydrolysis reaction time needed to obtain similar reducing sugar yield was significantly lower for microwave irradiation than shaker incubator. On the other hand, the reduction rate of the relative viscosity was significantly higher for the hydrolysis of chitosan using shaker incubator. A significant difference in chemical structure was observed between hydrolysis using microwave irradiation and shaker incubator. Overall, the result showed that the hydrolysis behavior of chitosan using microwave irradiation is significantly different with using shaker incubator.

Optimization of the Cellulase Free Xylanase Production by Immobilized Bacillus Pumilus.

BackgroundThe extracellular xylanase secreted by microorganisms is a hydrolytic enzyme, which arbitrarily cleaves the β-1, 4 backbone of the polysaccharide xylan; an enzyme used in the food processing, bio-pulping and bio-bleaching. The commercial production of the xylanase is limited because of a higher cost involvement, which can be overcome by the cost-effective production of the xylanase through immobilization of the microbial cell by the non-toxic substances.ObjectivesIn this work, the optimization of the extra-cellular cellulase free xylanase production by the immobilized cell of the Bacillus pumilus IMAU80221 strain using Ca-alginate beads along with standardization of the various parameters for a higher xylanase production were studied.Materials and MethodsFollowing to sterilization, the Na-alginate solution was mixed with the bacterial suspension of the Bacillus pumilus IMAU80221 and was added drop by drop into the 1 M calcium chloride solution for 1 h for obtaining a uniform sized polymeric bead of the Ca-alginate. For xylanase production, the Ca-alginate beads were then transferred into 100 mL Erlenmeyer flasks with 20 mL of the culture medium containing (w/v) 0.02% NaCl, 0.02% MgSO4, 0.04% (KH4)2PO4, 0.1% peptone, and 0.5% xylan and incubated at 34 °C in an incubator shaker (150 rpm) for 24 h. The resultant supernatant (crude enzyme) was used for enzyme assay.ResultsThe maximum xylanase production by the free cell (1.9 U.mL-1.min-1) was recorded at 48 h which was 40.5% lower than the xylanase production by the immobilized cell (2.67 U.mL-1.min-1) at the same time. The beads containing the immobilized cells could be reused up to eight fermentation cycles for xylanase production and retained 83.5% of the productivity at the fourth cycle. The entrapped cells were stable after six months of storage at 4 °C and retained 68% of the xylanase productivity.ConclusionCellulase free xylanase production from the immobilized Bacillus pumilus IMAU80221 was optimized. The xylanase production by the immobilized cells of Bacillus pumilus was higher by 40.5 and 132.6 % over the free cells respectively after 48 and 72 h of the incubation.

Effect of different nitrogen source combinations on microbial cellulose production by Pseudomonas aeruginosa in batch fermentation

Microbial cellulose (MC) is a type of polysaccharide that has high purity properties, which can be produced by different types of microorganism such as Pseudomonas, Komagataeibacter, Sarcina and Azotobacter and widely used in industrial applications. However, the main drawback in microbial cellulose production is low yield, which unable to meet the requirements of the industry. This study aims to determine the effects of different nitrogen sources combinations on microbial cellulose production by Pseudomonas aeruginosa and to maximize microbial cellulose production using the best nitrogen sources combination. To improve cellulose production, five different nitrogen sources combinations, which were yeast extract (YE) + (NH4)2SO4, YE + NH4NO3, YE + urea, YE + tryptic soy broth and YE + beef extract were added into the media with the composition of 50 g/L glucose, 3/L g KH2PO4 and 0.05 g/L MgSO4. The batch fermentation was done in incubator shaker with 150 rpm at 30°C for five days. The samples were harvested every 24 hours, centrifuged, purified and dried for microbial cellulose concentration determination. The results obtained indicated that YE and beef extract combination produced the highest microbial cellulose concentration of 1.7 g/L. To attain the second objective, optimization study was carried out by using different concentrations, which were 5 g/L, 10 g/L, 15 g/L and 20 g/L of YE and beef extract combination. An investigation using 10 g/L YE and beef extract has produced 1.7 g/L of microbial cellulose, which proved to be the potential enhancer for microbial cellulose synthesis. It is recommended that additional morphology analyses should be done to investigate the influence of fermentation conditions and low-cost nitrogen sources media components on microbial cellulose morphology and its mechanical properties. This will help to synthesize desired and reliable microbial cellulose pellicles.

English

Biosurfactants are surface-active compounds derived from microorganisms and offer several advantages over chemical surfactants, such as low toxicity, good biodegradability and ecological acceptability. Even though interest in biosurfactants is increasing, these bioproducts do not compete economically with synthetic surfactants due to the overall costs of the bioprocess. The use of inexpensive raw materials is an important approach to reduce these costs since the substrate price account for 10 to 30% of the final product expenses. In this study, papaya (Carica papaya L.) waste was used as a substrate to produce biosurfactant by Bacillus subtilis UFPEDA 86 strain. In addition, culture medium pH was corrected from 5.25 to 6.8 in order to analyze the influence of this variable on the biosurfactant production. The submerged fermentation was carried out on a shaker incubator at 37°C, 200 rpm for 96 h. Biomass and substrate concentration, surface tension, emulsification index and critical micelle concentration were analyzed. The strain was well adapted to both substrates studied, without and with pH correction. Using the broth without pH correction (pH=5.25), a maximum cell concentration of 1.07 g L-1 at 36 h a 25.5% surface tension reduction, emulsification index of around 61% and critical micelle concentration of about 35 mg L-1 was obtained. Using the broth with pH correction to 6.8, better results; maximum cell concentration of 1.14 g L-1 at 24 h, a 32.5% surface tension reduction, emulsification index of around 66% and critical micelle concentration at about 35 mg L-1 was obtained. The papaya waste proved to be an effective substrate in the biosurfactant production by B. subtilis UFPEDA 86 and the pH variable proved to be of great importance in the yield of this process. Key words: Biosurfactant, submerged fermentation, papaya waste, Bacillus subtilis.

Environmentally friendly dye for dye-sensitized solar cells from roots and stems of Berberis vulgaris

In this research, alkaloid compounds were extracted from the roots and stems of Berberis vulgaris using response surface methodology. The experimental design used absorption intensity as the response at three levels and four factors. The results showed that the factors of mesh and solvent percentage had more meaningful effects than those of temperature and time. Extractions were performed in an incubator shaker with 90% methanol solvent as optimal. The absorption intensity for the roots and stems of the plant was 1.299 and 0.867, respectively. The extracted dye was used in the fabrication of dye-sensitized solar cells. The surface morphology of the samples was studied using atomic force microscopy. Under optimal conditions, low aggregation between the dye molecules and TiO2 film decreased the average surface roughness in comparison with the condition in which the dye molecule concentration increased. The solar cell was built using optical electrodes sensitized with the roots and stems of B. vulgaris, and the calculated solar cell output under optimal conditions was 0.304 and 0.309, respectively. Electrochemical impedance spectroscopy was used to study the internal resistance of the solar cells, and it was shown that an increase in electron transfer between the dye molecule surface and TiO2 film decreased solar cell resistance.

Biodegradability of polyethylene by bacteria and fungi from Dandora dumpsite Nairobi-Kenya.

This study aimed at isolating and identifying bacteria and fungi with the capacity to degrade low density polyethylene (LDPE). The level of biodegradation of LDPE sheets with bacterial and fungal inoculums from different sampling points of Dandora dumpsite was evaluated under laboratory conditions. Incubation of the LDPE sheets was done for sixteen weeks at 37°C and 28°C for bacteria and fungi respectively in a shaker incubator. Isolation of effective candidates for biodegradation was done based on the recorded biodegradation outcomes. The extent of biodegradation on the polyethylene sheets was assessed by various techniques including weight loss analysis, Fourier Transform Infrared Spectroscopy (FTIR) and GC-MS. Fourier Transform Infra-Red spectroscopy (FTIR) analysis revealed the appearance of new functional groups attributed to hydrocarbon degradation after incubation with the bacteria and fungi. Analysis of the 16S rDNA and 18S rDNA sequences for bacteria and fungi respectively showed that bacteria belonging to genera Pseudomonas, Bacillus, Brevibacillus, Cellulosimicrobium, Lysinibacillus and fungi of genus Aspergillus were implicated as polyethylene degraders. An overall analysis confirmed that fungi are generally better degraders of polyethylene than bacteria. The highest fungal degradation activity was a mean weight reduction of 36.4±5.53% attributed to Aspergillus oryzae strain A5, 1 (MG779508). The highest degradation activity for bacteria was a mean of 35.72± 4.01% and 20.28± 2.30% attributed to Bacillus cereus strain A5,a (MG645264) and Brevibacillus borstelensis strain B2,2 (MG645267) respectively. Genus Aspergillus, Bacillus and Brevibacillus were confirmed to be good candidates for Low Density Poly Ethene bio-degradation. This was further confirmed by the appearance of the aldehyde, ether and carboxyl functional groups after FTIR analysis of the polythene sheets and the appearance of a ketone which is also an intermediary product in the culture media. To improve this degrading capacity through assessment of optimum conditions for microbial activity and enzyme production will enable these findings to be applied commercially and on a larger scale.

Lactic Acid Production From Cocoa Pod Husk by Studying Further the Influence of Alkaloids on Fermentation Process using Lactobacillus Plantarum Bacteria

Cocoa Pod Husk (CPH) is the biomass waste from cocoa industry, generated after the digestion process. It accounts for about 70-75% of the cocoa fresh fruit. CPH contains active compounds of alkaloids that are thought to be the inhibitors of the lactic acid fermentation process using microorganisms. This study aims to produce Lactic Acid from CPH by studying further the influence of alkaloids on fermentation process using Lactobacillus plantarum bacteria. Fermentation using Lactobacillus plantarum bacteria was carried out at 50°C and with agitation at 100 rpm in incubator shaker for 48 h. This process was conducted to determine the effect of the addition of alkaloids by analyzing through the comparison between the consumption of substrate (glucose), dry weight of the cell, and the production of lactic acid. Evaluation of the differences in the performance of microorganisms at various treatments was performed based on the parameters values of the kinetic models prepared for the case studied. The kinetic model fitting results showed that the presence of alkaloids alters the growth patterns of products from growth-associated products into mixed patterns because the products were formed during slow growth and stationary phases. The maximum growth rate (μm) and substrate inhibition constant (Ks) obtained on each variation of inhibitor addition were likely to remain constant at the values of 0.69 h-1 and 3.89 g/L respectively, as these parameters were unaffected by the addition of inhibitor

Bioconversion of acrylonitrile using nitrile hydratase activity of Bacillus sp. APB-6.

Bacillus sp. APB-6 harboring nitrile hydratase was used in the production of acrylamide from acrylonitrile. Bacillus sp. APB-6, for maximum production of Co++ containing nitrile hydratase, was cultured in the medium containing lactose (18.0gl-1), peptone (1.0gl-1), yeast extract (2.0gl-1), MgSO4 (0. 5gl-1), K2HPO4 (0.6gl-1), urea (9.0gl-1), and CoCl2 (0.01gl-1), pH 7.0, and incubated at 35°C for 24h in an incubator shaker (160rpm). Nitrile hydratase exhibited relatively high specificity for aliphatic nitriles. Free cells were immobilized using 2% (w/v) agar solution to enhance enzyme stability and reusability in repetitive cycles of acrylamide production. Under optimized conditions, nearly complete bioconversion of acrylonitrile was achieved with a fair recovery of 85% using free and immobilized cells equivalent to 500mg dcw l-1. An efficient nitrile hydratase-mediated bioconversion of acrylonitrile to acrylamide at 1-l scale was achieved with time and space productivity of 426gh-1 g-1 dcw using free cells.

PROTEASE FROM Bacillus sp. AS A DEGRADING CHICKEN FEATHER FOR PRODUCING FEATHER MEAL

Chicken feathers are a significant source of protein for livestock because ofhigh protein content and contain large amounts of cystine, glycine, arginine, and phenylalanine. Chicken feathers are not hydrolyzed, it is very difficult to digest by nonruminant animals because they contain high keratin. Hydrolyzed chicken feathers are not very difficult to digest by nonruminant animals because they contain high keratin. Raw chicken feathers must be hydrolyzed to be used as a feed ingredient for nonruminant species. Processing through bio-fermentation of chicken feathers would lead to overhaul the structure of the network, termination of disulfide bonds,hydrogen bonds and decreased levels of keratin. One of the processing of chicken feathers into feather meal is to utilize the specific proteases that degrade keratin. Bacillus sp. which has been isolated from Kawah Putih crater, Ciwidey, Bandung is a protease-producing bacterium. Fermentation using a medium containing: 2.8% (w/ v) glucose, 0.5% (w / v) urea and 0.005% (w / v) yeast extract with the conditions of the process carried out at pH 7.5; temperature 37 0C; shaker incubator 150 rpm will produce protease with the activity of 121 U / ml and prote in content 6.65 mg /ml in 36 hours. Hydrolysis of chicken feather needs to be pepsin digestibility tested to ensure that it has been processed correctly. Chicken feathers was obtained from slaughterhouses in Bogor. Pepsin digestibility of feather meal after hydrolyzed by proteases from Bacillus sp. is 67.4%.Keywords: feather meal, protease, fermentation, Bacillus sp.

Biodegradation of Anthracene Compound by Two Species of Filamentous Fungi

Isolation and identification fungi of Emericella nidulans and Aspergillus flavus from a pinkish and yellowish artificial clay, by using potato dextrose agar (PDA). Results revealed that E. nidulans was the best for degrading anthracene (92.3%) with maximum biomass production (3.7gm/l), compared to A. flavus with the rate of degradation (89%) and biomass production of (1.2gm/l), when methylene blue was used as redox indicator after incubating in a shaker incubator 120rpm at 30Co for 8days. Results indicated that E. nidulans has a high ability of anthracene degradation with the rate of (84%), while A. flavus showed the lower level with (77%) by using HPLC.

Green Synthesis of Silver Nanoparticles Using Hydnocarpus pentandra Leaf Extract: In-vitro Cyto-Toxicity Studies Against MCF-7 Cell Line

Objective: To synthesize silver nanoparticles from leaves extract of Hydnocarpus Pentandra (H. Pentandra ) and test the anti-cancer activity of synthesized silver nanoparticles against breast cancer cell line (MCF-7 cells). Method: Ten milliliter of leaves extract was mixed with 10 mL of aqueous 1 mM silver nitrate solution and kept in an incubator shaker system at 37oC for 24h. During this period, yellowish to reddish-brown color changes was observed. Characterization of silver nanoparticles using UV-vis spectrophotometry and SEM analysis were performed. Antioxidant activity of silver nanoparticles was performed using DPPH method. Anti-cancer activity of silver nanoparticles was tested using in vitro cytotoxicity test with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide against MCF-7 breast cancer cells. Results: UV-vis spectral analysis indicated the sliver surface plasmon resonance band at 438 nm. SEM analysis showed that silver nanoparticles to be spherical in shape with a size in the range of 141 nm to 202 nm. The synthesized silver nanoparticles exhibited antioxidant activity at dose dependent manner using DPPH method. The synthesized silver nanoparticles efficiently reduced the viability of MCF-7 breast cancer cells in a dose dependent manner. Conclusion: It has been demonstrated that the leaves extract of Hydnocarpus Pentandra could produce silver nanoparticles and the silver nanoparticles are quite stable in solution. Further studies are needed to characterize the mechanism involved with anti-cancer activity of the silver nanoparticles. Key words: Hydnocarpus Pentandra, Leaf, Anti-Oxidant, Anti-Cancer Activity.

Petroleum residues degradation in laboratory-scale by rhizosphere bacteria isolated from the mangrove ecosystem

This research is about petroleum bioremediation experiment to obtain bacterial isolate from mangrove ecosystem which potentially degrade petroleum. It was conducted in an Erlenmeyer batch system filled with growth medium of Stone Mineral Salt Solution (SMSS) plus petroleum residue, placed in an incubator shaker with a rotation speed of 120 rpm, temperature 3000C, for 14 research days. Indigenous bacteria that have been isolated and identified from the roots of mangrove plants are Ochrobactrum anthropi and Bacillus sp., Ralstonia pickettii and Bacillus circulans. Those bacteriain both monoculture and consortium form (mixed culture) are incorporated into erlenmeyer as remediator agents. All bacteria can utilize hydrocarbon compounds, but Ralstonia pickettii and Bacillus circulans reached exponential phase faster with more cell count than other bacteria. Compared to single cultures, petroleum degradation by a bacterial consortium provides a higher TPH reduction efficiency, i.e. at 5%, 10%, and 15% of initial TPH of 94.4%, 72%, and 80.3%, respectively. This study proved that all bacteria could optimize hydrocarbon compounds up to 15% TPH load.

Evaluation of the antimicrobial potential of sulfated polysaccharides from the marine invasive algae Asparagopsis armata (Harvey, 1855).

Event Abstract Back to Event Evaluation of the antimicrobial potential of sulfated polysaccharides from the marine invasive algae Asparagopsis armata (Harvey, 1855). Fernando Teodoro1, Fábio Gomes1, Susete Pinteus1, Celso Alves1, Joana Silva1, Susana M. Bernardino1 and Rui P. Pedrosa1* 1 MARE – Marine and Environmental Sciences Centre, ESTM, Polytechnic Institute of Leiria, Portugal The marine environment harbors a high number of ecosystems, where a wide diversity of organisms capable of producing interesting secondary metabolites can be found. Among these organisms are the macroalgae, sources of renewable resources characterized as being rich in biological properties [1]. Among other metabolites, these present sulfated polysaccharides (SPs) in the constitution of their cell wall, molecules widely studied for their biotechnological potential. Asparagopsis armata (Harvey, 1855) is an invading seaweed native to the Australian coast that has been spreading throughout the Portuguese coast [2]. The invasive status of this algae and low economical value, potentiate the search for novel compelling applications, valorizing this specie. Accordingly, this study evaluates the antimicrobial properties of SPs, extracted from Asparagopsis armata collected in the Peniche coast, against Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), Candida albicans (ATCC 10231), Pseudomonas aeruginosa (ATCC 27853) and Salmonella enterica (ATCC 13076) through growth inhibition analysis. The growth of E. coli, S. aureus, P. aeruginosa and S. enterica was performed in LB medium while the growth of C. albicans was performed in yeast-extract-peptone-dextrose, placed in a shaker incubator at a temperature of 37 °C until reach exponential growth (OD600). For the evaluation of the inhibitory capacity, the microorganisms were subjected to SP samples at a concentration range of 10 – 500 µg mL-1. The inhibitory effects were most expressive against S. aureus and P. aeruginosa, with an IC50 of 31.51 (25.29-39.26) µg mL-1 and 309.90 (248.00 – 384.90) µg mL-1, respectively. All statistical analysis was performed using GraphPad Prism (Version 7.04) software for windows. Our results demonstrate that Asparagopsis armata presents a potential applicability as antibacterial against S. aureus and P. aeruginosa, thus being able to grant some economic value from this invasive seaweed. Acknowledgements Authors would like to acknowledge the financial support of the Portuguese Foundation for Science and Technology (FCT) through Strategic Project UID/MAR/04292/2013 granted to MARE – Marine and Environmental Sciences Centre, through Red2Discovery Project (PTDC/MAR-BIO/6149/2014), co-financed by COMPETE (POCI-01-0145-FEDER-016791), and through Oncologia de Precisão: Terapias e Tecnologias Inovadoras project (POINT4PAC) (SAICTPAC/0019/2015 - LISBOA-01-0145-FEDER-016405).

Biodegradation of Anthracene Compound by Two Species of Filamentous Fungi

Isolation and identification fungi of Emericella nidulans and Aspergillus flavus from a pinkish and yellowish artificial clay, by using potato dextrose agar (PDA). Results revealed that E. nidulans was the best for degrading anthracene (92.3%) with maximum biomass production (3.7gm/l), compared to A. flavus with the rate of degradation (89%) and biomass production of (1.2gm/l), when methylene blue was used as redox indicator after incubating in a shaker incubator 120rpm at 30Co for 8days. Results indicated that E. nidulans has a high ability of anthracene degradation with the rate of (84%), while A. flavus showed the lower level with (77%) by using HPLC.