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Neural cell chip to detect neurotoxin substance in contaminated water
Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576
calculated, and correlation matrix analysis was performed to determining the correlation of molecular properties and degradation. The results showed that potential energy, molecular orbital energy, and hydrophobicity of PCB substrates could influence interactions of BDO and PCB. The results of molecular simulation indicated that the model of BDO from Enterobacter sp. LY402 obtained by homology model was reasonable and reliable, and the properties of different PCB substrates played a significant role in inclusion and catalyzing by BDO. doi:10.1016/j.jbiotec.2010.09.074 [P-E.53] Optimization of immobilization conditions for petroleum oil biodegradation using wood chips and wax as carrier for Candida spp. Soha Farag ∗ , Yasser R. Abdel-Fattah, Nadia A. Soliman Mubarak City for Scientific Research and Technology Applications, Egypt Keywords: Biodegradation; carrier; semi-continuous fermentation cycle; yeast strains Water contamination with crude oil from petrochemicals and oil exploitation is an important worldwide issue. Comparing available remediation techniques, bioremediation is widely considered to be a cost effective choice. To promote crude oil removal, biocarrier for immobilization of indigenous hydrocarbon-degrading bacteria was developed using porous materials such as wood chips and wax. In this study two yeast (designated as AQ1 & AQ2) were isolated from motor oil polluted area of Abou-Qir gulf (Alexandria, Egypt) and selected among group of environmental isolates, due to its potential activity in petroleum degradation. Based on partial sequences of 18Sr RNA gene homology, the strain AQ1 was 99% related to Candida sp. S42-12 (EC.EF498418). And strain AQ2 was 99% related to Candida tropicalis strain zhuan 143 (EC. EF 428133) according to phylogenetic tree. The biodegradation potential increased with increasing the time of strain incubation with the carrier from 1hr-5hr. when using a woodchips as a carrier 1.75 g/44 cm2 surface area of the flask, the percentage of crude oil removal reached to 90% and 73% for stains AQ1 and AQ2 respectively. While in case of wax the percentage reached to 70% and 62% for strains AQ1 and AQ2 respectively, using an 11 g/44 cm2 surface area. High microbial colonization by spherical and rod shapes were observed for the 1-3 mm thick biofilm on the outer surface of biocarrier (wood chip) using scanning electronic microscopy (SEM). Based on batch-scale experiments containing free-living bacterial cultures and biocarrier into crude oil contaminated water, biocarrier enhanced the biodegradation of crude oil, with different degree according to different strains and to different carriers. A new semi-continuous fermentation cycle was done in which further immobilization takes place. This study explored the role of biocarrier in enhancing biodegradation of hydrophobic contaminants such as crude oil.
S231
[P-E.54] Isolation and molecular characterization of some copper biosorped strains Sahar Zaki, Soha farag ∗ Mubarak City for Scientific Research and Technology Applications, Egypt Keywords: Biosorption; Copper; RFLP; Bioluminescent Heavy metals are groups of pollutants, which are not biodegradable and tend to accumulate in living organisms. The concentrations of these metals need to be reduced to meet ever-changing legislative standards. The biosorption of Cu(II) from aqueous solution using different bacterial strains was studied. Copper-biosorbing bacteria were isolated from tannery effluent collected from Borg AlArab, Alexandria, Egypt. These isolates displayed different degrees of Copper biosorption under aerobic conditions. Based on 16S rDNA gene sequence analysis, three of them (S2, S5, and S7) were identified as Chryseobacterium sp., Enterobacter sp., and Stenotrophomonas sp., respectively. Initial Cu(II) ion concentrations from 25-250 mg/l at constant temperature 30 ◦ C were studied. The residual Cu(II) concentration and its toxicity effect in solution were determined using atomic absorption spectrophotometer and bioluminescent bioreporter. The maximum biosorption efficiency of 225 mg/l Cu by isolates S2, S5 and S7 was 48%, 71% and 70.1%, respectively. The bioluminescence inhibition of strain (S5) reached to 91.4% as compared with the strain (S7) reached to 83.3% at 225 mg/l of Cu ion. The bioluminescent bioreporter was proved to be fast and accurate technique for measurement the toxicity effect of residual Cu(II) in solution. doi:10.1016/j.jbiotec.2010.09.076 [P-E.55] Neural cell chip to detect neurotoxin substance in contaminated water J. Park, Y.-K. Kim ∗ Hankyong National University, Republic of Korea Keywords: Neural cell chip; microfluidic chip; Neurotoxin; microvalve Neurotoxin substances in environment show extremely harmful effect to nervous system of human being and animal in ecosystem. The sensitive, correct, and rapid monitoring of those neurotoxin substances is very important to prevent ecosystem crisis. In this study, a neural cell chip using the microvalve integrated microfluidic device was developed to detect neurotoxin substances. Fig. 1. Neural cell chip was composed of microfluidic device and microelectrode patterned glass plate. Microfluidic device has an
doi:10.1016/j.jbiotec.2010.09.075
Fig. 1. Images of (a) a neural cell chip and (b) a chamber with PC-12 cells.
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Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576
integrated microvalve system and made by polydimethylsiloxane (PDMS). Neural cells (PC12) were cultivated in a chamber of microfluidic device and samples with neurotoxin substances were applied into a chamber via micro-channel. The electric property according to concentration of dopamine, which is excreted from PC12 cells, was measured to obtain signal. In this work, several concentrations of neurotoxin were used to make a calibration curve between obtained signal and concentration. A proposed neural cell chip can successfully detect neurotoxin substances at various concentration. Acknowledgement: This study was supported by Ministry of Environment as “The Eco-technopia 21 project”. Fig. 1. – Oxidation of 1 mM of As(III) to As(V) during the growth of the isolated As(III)-resistant bacterial strain SCR II 1a.
doi:10.1016/j.jbiotec.2010.09.077 [P-E.56]
[P-E.57]
Arsenite [As(III)] oxidation to arsenate [As(V)] in heterotrophic bacterial strains isolated from the dump site of Scarlino, Tuscany, Italy E. Bernardini 1 , S. Focardi 2 , V Ruta 1,∗ , M. Pepi 1 , S.E. Focardi 1
Niccolucci 2 , S.
Gasperini 3 , M.
1
University of Siena Department of Environmental Sciences, Italy University of Siena Department of Chemistry, Italy 3 University of Siena Faculty of Sciences, Italy Keywords: arsenic; bioremediation; arsenite; oxidation 2
Arsenic is a metalloid widely distributed in the environment, released both from natural and anthropogenic sources. The most common oxidation states of arsenic in the environment are arsenate As(V) and arsenite As(III). Of these two forms, As(III) is more toxic. Several remediation processes have been described for arsenic removal based on chemical oxidation of As(III) to As(V) followed by alkaline precipitation. These processes generate additional pollution and are expensive, thus the need to experiment with alternative methods based on its biological oxidation. Sixteen bacterial strains were isolated from an arsenic contaminated dump site in Tuscany, Italy, in the presence of As(III) 13.5 mM, added as m-NaAsO2 . Values of minimum inhibitory concentrations (MICs) in the presence of As(III) showed highest levels of 6.8 and 13.5 mM in the isolates. Concerning MICs in the presence of As(V), eight out of sixteen bacterial strains thrived in the presence of 13.5 mM of As(V) added as Na2 HAsO4 .7H2 O. Several isolates showed the capability to reduce 2 mM of As(V) to As(III) within 24 – 48 hours and, interestingly, two bacterial strains showed a mechanism of resistance based on the oxidation of As(III) to As(V), the less toxic form of this metalloid and less soluble, as respect to As(III). In particular, strain SCR II 1a was able to oxidize 1 mM of As(III) within 24 hours (Fig. 1), resulting one of the most promising isolates to be used in As(III) bioremediation processes. The oxidizing activity of this bacterial strain was confirmed testing its capability to oxidize As(III) to As(V) on solid medium, by using a solution of AgNO3 to reveal the two forms of arsenic. Studies on the aox genes are in progress. This study suggests that the As(III)-oxidizing bacterial strain SCR II 1a could have a role in possible bioremediation processes in sites contaminated by arsenic. doi:10.1016/j.jbiotec.2010.09.078
Biosurfactant production by heterotrophic bacteria isolated from aquaculture plant wastes and their growth in the presence of oleic acid as sole carbon and energy source A. Lobianco 1,∗ , D.L. Angelini 1 , F. Borghini 1 , S. Focardi 2 , M. Pepi 1 , S.E. Focardi 1 1
University of Siena Department of Environmental Sciences, Via P.A. Mattioli, 4; 53100 Siena, Italy 2 University of Siena Department of Chemistry, Via A. Moro, 2; 53100 Siena, Italy Keywords: aquaculture; beneficial microorganisms; biosurfactant; biodegradation In intensive aquaculture, fish are under high-density stress, and fishponds with lots of humus (remnants of feeds and feces) are causing the fish to be susceptible to diseases. Therefore, it is extremely important to supplement beneficial microbes (BMs) to speed up the decomposition of organic matter in water and sediments so as to maintain good water ambience. Degradation of hydrophobic substrata can be enhanced by biosurfactants produced by microorganisms. Seven bacterial strains were isolated from intensive aquaculture wastes, in the presence of 2% of oleic acid, a component of wastes, as the sole carbon and energy source. The isolated bacterial strains were characterized from the microbiological point of view revealing rod, Gram negative cells. Isolates showed degradation both of oleic acid and of native aquaculture wastes. Microbial adhesion test to hydrocarbons (MATH) showed that the isolates were constitutional hydrophobic. Growth on blood agar highlighted haemolysis in all the isolates and the production of biosurfactants was further investigated. Emulsification test (E24%) showed an increase of emulsions originated by the biosurfactant and hydrocarbons, along with incubation of the bacterial strains in the presence of oleic acid. Among the isolates, strain DG2a showed the best growth in the presence of oleic acid (Fig. 1a), showing a good efficiency in producing biosurfactant. An increase of haloes of degradation was also detected in the DG2a bacterial strain in the presence of oleic acid, as detected by measuring the diameter of clear zone determined by OST and the emulsification activity (E24%) was good (Fig. 1b). Production of biosurfactant by this isolated bacterial strain could help aquaculture wastes degradation. The bacterial strains isolated in this study could be useful BMs in enhancing degradation of rich in humus aquaculture wastes con-
calculated, and correlation matrix analysis was performed to determining the correlation of molecular properties and degradation. The results showed that potential energy, molecular orbital energy, and hydrophobicity of PCB substrates could influence interactions of BDO and PCB. The results of molecular simulation indicated that the model of BDO from Enterobacter sp. LY402 obtained by homology model was reasonable and reliable, and the properties of different PCB substrates played a significant role in inclusion and catalyzing by BDO. doi:10.1016/j.jbiotec.2010.09.074 [P-E.53] Optimization of immobilization conditions for petroleum oil biodegradation using wood chips and wax as carrier for Candida spp. Soha Farag ∗ , Yasser R. Abdel-Fattah, Nadia A. Soliman Mubarak City for Scientific Research and Technology Applications, Egypt Keywords: Biodegradation; carrier; semi-continuous fermentation cycle; yeast strains Water contamination with crude oil from petrochemicals and oil exploitation is an important worldwide issue. Comparing available remediation techniques, bioremediation is widely considered to be a cost effective choice. To promote crude oil removal, biocarrier for immobilization of indigenous hydrocarbon-degrading bacteria was developed using porous materials such as wood chips and wax. In this study two yeast (designated as AQ1 & AQ2) were isolated from motor oil polluted area of Abou-Qir gulf (Alexandria, Egypt) and selected among group of environmental isolates, due to its potential activity in petroleum degradation. Based on partial sequences of 18Sr RNA gene homology, the strain AQ1 was 99% related to Candida sp. S42-12 (EC.EF498418). And strain AQ2 was 99% related to Candida tropicalis strain zhuan 143 (EC. EF 428133) according to phylogenetic tree. The biodegradation potential increased with increasing the time of strain incubation with the carrier from 1hr-5hr. when using a woodchips as a carrier 1.75 g/44 cm2 surface area of the flask, the percentage of crude oil removal reached to 90% and 73% for stains AQ1 and AQ2 respectively. While in case of wax the percentage reached to 70% and 62% for strains AQ1 and AQ2 respectively, using an 11 g/44 cm2 surface area. High microbial colonization by spherical and rod shapes were observed for the 1-3 mm thick biofilm on the outer surface of biocarrier (wood chip) using scanning electronic microscopy (SEM). Based on batch-scale experiments containing free-living bacterial cultures and biocarrier into crude oil contaminated water, biocarrier enhanced the biodegradation of crude oil, with different degree according to different strains and to different carriers. A new semi-continuous fermentation cycle was done in which further immobilization takes place. This study explored the role of biocarrier in enhancing biodegradation of hydrophobic contaminants such as crude oil.
S231
[P-E.54] Isolation and molecular characterization of some copper biosorped strains Sahar Zaki, Soha farag ∗ Mubarak City for Scientific Research and Technology Applications, Egypt Keywords: Biosorption; Copper; RFLP; Bioluminescent Heavy metals are groups of pollutants, which are not biodegradable and tend to accumulate in living organisms. The concentrations of these metals need to be reduced to meet ever-changing legislative standards. The biosorption of Cu(II) from aqueous solution using different bacterial strains was studied. Copper-biosorbing bacteria were isolated from tannery effluent collected from Borg AlArab, Alexandria, Egypt. These isolates displayed different degrees of Copper biosorption under aerobic conditions. Based on 16S rDNA gene sequence analysis, three of them (S2, S5, and S7) were identified as Chryseobacterium sp., Enterobacter sp., and Stenotrophomonas sp., respectively. Initial Cu(II) ion concentrations from 25-250 mg/l at constant temperature 30 ◦ C were studied. The residual Cu(II) concentration and its toxicity effect in solution were determined using atomic absorption spectrophotometer and bioluminescent bioreporter. The maximum biosorption efficiency of 225 mg/l Cu by isolates S2, S5 and S7 was 48%, 71% and 70.1%, respectively. The bioluminescence inhibition of strain (S5) reached to 91.4% as compared with the strain (S7) reached to 83.3% at 225 mg/l of Cu ion. The bioluminescent bioreporter was proved to be fast and accurate technique for measurement the toxicity effect of residual Cu(II) in solution. doi:10.1016/j.jbiotec.2010.09.076 [P-E.55] Neural cell chip to detect neurotoxin substance in contaminated water J. Park, Y.-K. Kim ∗ Hankyong National University, Republic of Korea Keywords: Neural cell chip; microfluidic chip; Neurotoxin; microvalve Neurotoxin substances in environment show extremely harmful effect to nervous system of human being and animal in ecosystem. The sensitive, correct, and rapid monitoring of those neurotoxin substances is very important to prevent ecosystem crisis. In this study, a neural cell chip using the microvalve integrated microfluidic device was developed to detect neurotoxin substances. Fig. 1. Neural cell chip was composed of microfluidic device and microelectrode patterned glass plate. Microfluidic device has an
doi:10.1016/j.jbiotec.2010.09.075
Fig. 1. Images of (a) a neural cell chip and (b) a chamber with PC-12 cells.
S232
Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576
integrated microvalve system and made by polydimethylsiloxane (PDMS). Neural cells (PC12) were cultivated in a chamber of microfluidic device and samples with neurotoxin substances were applied into a chamber via micro-channel. The electric property according to concentration of dopamine, which is excreted from PC12 cells, was measured to obtain signal. In this work, several concentrations of neurotoxin were used to make a calibration curve between obtained signal and concentration. A proposed neural cell chip can successfully detect neurotoxin substances at various concentration. Acknowledgement: This study was supported by Ministry of Environment as “The Eco-technopia 21 project”. Fig. 1. – Oxidation of 1 mM of As(III) to As(V) during the growth of the isolated As(III)-resistant bacterial strain SCR II 1a.
doi:10.1016/j.jbiotec.2010.09.077 [P-E.56]
[P-E.57]
Arsenite [As(III)] oxidation to arsenate [As(V)] in heterotrophic bacterial strains isolated from the dump site of Scarlino, Tuscany, Italy E. Bernardini 1 , S. Focardi 2 , V Ruta 1,∗ , M. Pepi 1 , S.E. Focardi 1
Niccolucci 2 , S.
Gasperini 3 , M.
1
University of Siena Department of Environmental Sciences, Italy University of Siena Department of Chemistry, Italy 3 University of Siena Faculty of Sciences, Italy Keywords: arsenic; bioremediation; arsenite; oxidation 2
Arsenic is a metalloid widely distributed in the environment, released both from natural and anthropogenic sources. The most common oxidation states of arsenic in the environment are arsenate As(V) and arsenite As(III). Of these two forms, As(III) is more toxic. Several remediation processes have been described for arsenic removal based on chemical oxidation of As(III) to As(V) followed by alkaline precipitation. These processes generate additional pollution and are expensive, thus the need to experiment with alternative methods based on its biological oxidation. Sixteen bacterial strains were isolated from an arsenic contaminated dump site in Tuscany, Italy, in the presence of As(III) 13.5 mM, added as m-NaAsO2 . Values of minimum inhibitory concentrations (MICs) in the presence of As(III) showed highest levels of 6.8 and 13.5 mM in the isolates. Concerning MICs in the presence of As(V), eight out of sixteen bacterial strains thrived in the presence of 13.5 mM of As(V) added as Na2 HAsO4 .7H2 O. Several isolates showed the capability to reduce 2 mM of As(V) to As(III) within 24 – 48 hours and, interestingly, two bacterial strains showed a mechanism of resistance based on the oxidation of As(III) to As(V), the less toxic form of this metalloid and less soluble, as respect to As(III). In particular, strain SCR II 1a was able to oxidize 1 mM of As(III) within 24 hours (Fig. 1), resulting one of the most promising isolates to be used in As(III) bioremediation processes. The oxidizing activity of this bacterial strain was confirmed testing its capability to oxidize As(III) to As(V) on solid medium, by using a solution of AgNO3 to reveal the two forms of arsenic. Studies on the aox genes are in progress. This study suggests that the As(III)-oxidizing bacterial strain SCR II 1a could have a role in possible bioremediation processes in sites contaminated by arsenic. doi:10.1016/j.jbiotec.2010.09.078
Biosurfactant production by heterotrophic bacteria isolated from aquaculture plant wastes and their growth in the presence of oleic acid as sole carbon and energy source A. Lobianco 1,∗ , D.L. Angelini 1 , F. Borghini 1 , S. Focardi 2 , M. Pepi 1 , S.E. Focardi 1 1
University of Siena Department of Environmental Sciences, Via P.A. Mattioli, 4; 53100 Siena, Italy 2 University of Siena Department of Chemistry, Via A. Moro, 2; 53100 Siena, Italy Keywords: aquaculture; beneficial microorganisms; biosurfactant; biodegradation In intensive aquaculture, fish are under high-density stress, and fishponds with lots of humus (remnants of feeds and feces) are causing the fish to be susceptible to diseases. Therefore, it is extremely important to supplement beneficial microbes (BMs) to speed up the decomposition of organic matter in water and sediments so as to maintain good water ambience. Degradation of hydrophobic substrata can be enhanced by biosurfactants produced by microorganisms. Seven bacterial strains were isolated from intensive aquaculture wastes, in the presence of 2% of oleic acid, a component of wastes, as the sole carbon and energy source. The isolated bacterial strains were characterized from the microbiological point of view revealing rod, Gram negative cells. Isolates showed degradation both of oleic acid and of native aquaculture wastes. Microbial adhesion test to hydrocarbons (MATH) showed that the isolates were constitutional hydrophobic. Growth on blood agar highlighted haemolysis in all the isolates and the production of biosurfactants was further investigated. Emulsification test (E24%) showed an increase of emulsions originated by the biosurfactant and hydrocarbons, along with incubation of the bacterial strains in the presence of oleic acid. Among the isolates, strain DG2a showed the best growth in the presence of oleic acid (Fig. 1a), showing a good efficiency in producing biosurfactant. An increase of haloes of degradation was also detected in the DG2a bacterial strain in the presence of oleic acid, as detected by measuring the diameter of clear zone determined by OST and the emulsification activity (E24%) was good (Fig. 1b). Production of biosurfactant by this isolated bacterial strain could help aquaculture wastes degradation. The bacterial strains isolated in this study could be useful BMs in enhancing degradation of rich in humus aquaculture wastes con-