Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1179

Search results for: thermophilic bacteria

1179 Potential Use of Spore-Forming Biosurfactant Producing Bacteria in Oil-Pollution Bioremediation

Authors: S. N. Al-Bahry, Y. M. Al-Wahaibi, S. J. Joshi, E. A. Elshafie, A. S. Al-Bimani


Oman is one of the oil producing countries in the Arabian Peninsula and the Gulf region. About 30-40 % of oil produced from the Gulf is transported globally along the seacoast of Oman. Oil pollution from normal tanker operations, ballast water, illegal discharges and accidental spills are always serious threats to terrestrial and marine habitats. Due to Oman’s geographical location at arid region where the temperature ranges between high 40s and low 50s Celsius in summers with low annual rainfall, the main source of fresh water is desalinated sea and brackish water. Oil pollution, therefore, pose a major threat to drinking water. Biosurfactants are secondary metabolites produced by microorganisms in hydrophobic environments to release nutrients from solid surfaces, such as oil. In this study, indigenous oil degrading thermophilic spore forming bacteria were isolated from oil fields contaminated soil. The isolates were identified using MALDI-TOF biotyper and 16s RNA. Their growth conditions were optimized for the production of biosurfactant. Surface tension, interfacial tensions and microbial oil biodegradation capabilities were tested. Some thermophilic bacteria degraded either completely or partially heavy crude oil (API 10-15) within 48h suggesting their high potential in oil spill bioremediation and avoiding the commonly used physical and chemical methods which usually lead to other environmental pollution.

Keywords: bacteria, bioremediation, biosurfactant, crude-oil-pollution

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1178 Fermentation of Pretreated Herbaceous Cellulosic Wastes to Ethanol by Anaerobic Cellulolytic and Saccharolytic Thermophilic Clostridia

Authors: Lali Kutateladze, Tamar Urushadze, Tamar Dudauri, Besarion Metreveli, Nino Zakariashvili, Izolda Khokhashvili, Maya Jobava


Lignocellulosic waste streams from agriculture, paper and wood industry are renewable, plentiful and low-cost raw materials that can be used for large-scale production of liquid and gaseous biofuels. As opposed to prevailing multi-stage biotechnological processes developed for bioconversion of cellulosic substrates to ethanol where high-cost cellulase preparations are used, Consolidated Bioprocessing (CBP) offers to accomplish cellulose and xylan hydrolysis followed by fermentation of both C6 and C5 sugars to ethanol in a single-stage process. Syntrophic microbial consortium comprising of anaerobic, thermophilic, cellulolytic, and saccharolytic bacteria in the genus Clostridia with improved ethanol productivity and high tolerance to fermentation end-products had been proposed for achieving CBP. 65 new strains of anaerobic thermophilic cellulolytic and saccharolytic Clostridia were isolated from different wetlands and hot springs in Georgia. Using new isolates, fermentation of mechanically pretreated wheat straw and corn stalks was done under oxygen-free nitrogen environment in thermophilic conditions (T=550C) and pH 7.1. Process duration was 120 hours. Liquid and gaseous products of fermentation were analyzed on a daily basis using Perkin-Elmer gas chromatographs with flame ionization and thermal detectors. Residual cellulose, xylan, xylose, and glucose were determined using standard methods. Cellulolytic and saccharolytic bacteria strains degraded mechanically pretreated herbaceous cellulosic wastes and fermented glucose and xylose to ethanol, acetic acid and gaseous products like hydrogen and CO2. Specifically, maximum yield of ethanol was reached at 96 h of fermentation and varied between 2.9 – 3.2 g/ 10 g of substrate. The content of acetic acid didn’t exceed 0.35 g/l. Other volatile fatty acids were detected in trace quantities.

Keywords: anaerobic bacteria, cellulosic wastes, Clostridia sp, ethanol

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1177 Isolation and Characterization of Lactic Acid Bacteria from Libyan Traditional Fermented Milk "Laban"

Authors: M. H. Nahaisi, N. M. Almaroum


Laban is a Libyan traditional fermented milk product. This lactic fermentation has been known in many cities of Libya long time ago as stable, nutritious, refreshing drink especially during the summer. 16 naturally fermented milk samples were collected from different cities located in North West of Libya. The average pH, titratable acidity, fat and total solids were 4.16, 0.73%, 1.54% and 8.12 % respectively. Coliform, yeast and mold counts were 21×10⁴, 39×10⁴ and 41 ×10³ cfu/ ml. respectively. The average Lactococcus, Streptococcus, Mesophilic Lactobacillus / Leuconostoc and Thermophilic Lactobacillus counts were 99 ×10⁷, 96 ×10⁷, 93 ×10⁷ and 15 ×10⁷ cfu / ml. respectively. A total of one hundred forty two lactic acid bacteria (LAB) isolates were identified to the genus level as Lactobacillus (48.59%), Lactococcus (43.66%), Streptococcus (4.93%) and Leuconostoc (2.82%). Sugar fermentation tests have revealed that the most frequently Lactobacillus species was found to be Lactobacillus delbrueckii ssp. lactis (62.32%) followed by Lactobacillus plantarum (31.88%). Furthermore, other selected LAB isolates were identified by API 50 CH test as Lactococcus lactis ssp. lactics, Lactobacillus pentosus, Lactobacillus brevis and Leuconostoc mesenteroides ssp. cremoris.

Keywords: traditional fermented milk, laban, lactococcus, streptococcus, mesophilic lactobacillus, thermophilic lactobacillus counts

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1176 Modelling and Simulation of a Commercial Thermophilic Biogas Plant

Authors: Jeremiah L. Chukwuneke, Obiora E. Anisiji, Chinonso H. Achebe, Paul C. Okolie


This paper developed a mathematical model of a commercial biogas plant for urban area clean energy requirement. It identified biodegradable waste materials like domestic/city refuse as economically viable alternative source of energy. The mathematical formulation of the proposed gas plant follows the fundamental principles of thermodynamics, and further analyses were accomplished to develop an algorithm for evaluating the plant performance preferably in terms of daily production capacity. In addition, the capacity of the plant is equally estimated for a given cycle of operation and presented in time histories. A nominal 1500 m3 power gas plant was studied characteristically and its performance efficiency evaluated. It was observed that the rate of bio gas production is essentially a function of the reactor temperature, pH, substrate concentration, rate of degradation of the biomass, and the accumulation of matter in the system due to bacteria growth. The results of this study conform to a very large extent with reported empirical data of some existing plant and further model validations were conducted in line with classical records found in literature.

Keywords: energy and mass conservation, specific growth rate, thermophilic bacteria, temperature, rate of bio gas production

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1175 Bioremediation of Arsenic from Industrially Polluted Soil of Vatva, Ahmedabad, Gujarat, India

Authors: C. Makwana, S. R. Dave


Arsenic is toxic to almost all living cells. Its contamination in natural sources affects the growth of microorganisms. The presence of arsenic is associated with various human disorders also. The attempt of this sort of study provides information regarding the performance of our isolated microorganisms in the presence of Arsenic, which have ample scope for bioremediation. Six isolates were selected from the polluted sample of industrial zone Vatva, Ahmedabad, Gujarat, India, out of which two were Thermophilic organisms. The thermophilic exopolysaccharide (EPS) producing Bacillus was used for microbial enhance oil recovery (MEOR) and in the bio beneficiation. Inorganic arsenic primarily exists in the form of arsenate or arsenite. This arsenic resistance isolate was capable of transforming As +3 to As+5. This isolate would be useful for arsenic remediation standpoint from aquatic systems. The study revealed that the thermophilic microorganism was growing at 55 degree centigrade showed considerable remediation property. The results on the growth and enzyme catalysis would be discussed in response to Arsenic remediation.

Keywords: aquatic systems, thermophilic, exopolysacchride, arsenic

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1174 Anti Staphylococcus aureus and Methicillin Resistant Staphylococcus aureus Action of Thermophilic Fungi Acrophialophora levis IBSD19 and Determination of Its Mode of Action Using Electron Microscopy

Authors: Shivankar Agrawal, Indira Sarangthem


Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA) remains one of the major causes of healthcare-associated and community-onset infections worldwide. Hence the search for non-toxic natural compounds having antibacterial activity has intensified for future drug development. The exploration of less studied niches of Earth can highly increase the possibility to discover novel bioactive compounds. Therefore, in this study, the cultivable fraction of fungi from the sediments of natural hot springs has been studied to mine potential fungal candidates with antibacterial activity against the human pathogen Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. We isolated diverse strains of thermophilic fungi from a collection of samples from sediment. Following a standard method, we isolated a promising thermophilic fungus strain IBSD19, identified as Acrophialophora levis, possessing the potential to produce an anti-Staphylococcus aureus agent. The growth conditions were optimized and scaled to fermentation, and its produced extract was subjected to chemical extraction. The ethyl acetate fraction was found to display significant activity against Staphylococcus aureus and MRSA with a minimum inhibitory concentration (MIC) of 0.5 mg/ml and 4 mg/ml, respectively. The cell membrane integrity assay and SEM suggested that the fungal metabolites cause bacteria clustering and further lysis of the cell.

Keywords: antibacterial activity, antioxidant, fungi, Staphylococcus aureus, MRSA, thermophiles

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1173 Selection Effects on the Molecular and Abiotic Evolution of Antibiotic Resistance

Authors: Abishek Rajkumar


Antibiotic resistance can occur naturally given the selective pressure placed on antibiotics. Within a large population of bacteria, there is a significant chance that some of those bacteria can develop resistance via mutations or genetic recombination. However, a growing public health concern has arisen over the fact that antibiotic resistance has increased significantly over the past few decades. This is because humans have been over-consuming and producing antibiotics, which has ultimately accelerated the antibiotic resistance seen in these bacteria. The product of all of this is an ongoing race between scientists and the bacteria as bacteria continue to develop resistance, which creates even more demand for an antibiotic that can still terminate the newly resistant strain of bacteria. This paper will focus on a myriad of aspects of antibiotic resistance in bacteria starting with how it occurs on a molecular level and then focusing on the antibiotic concentrations and how they affect the resistance and fitness seen in bacteria.

Keywords: antibiotic, molecular, mutation, resistance

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1172 High Efficient Biohydrogen Production from Cassava Starch Processing Wastewater by Two Stage Thermophilic Fermentation and Electrohydrogenesis

Authors: Peerawat Khongkliang, Prawit Kongjan, Tsuyoshi Imai, Poonsuk Prasertsan, Sompong O-Thong


A two-stage thermophilic fermentation and electrohydrogenesis process was used to convert cassava starch processing wastewater into hydrogen gas. Maximum hydrogen yield from fermentation stage by Thermoanaerobacterium thermosaccharolyticum PSU-2 was 248 mL H2/g-COD at optimal pH of 6.5. Optimum hydrogen production rate of 820 mL/L/d and yield of 200 mL/g COD was obtained at HRT of 2 days in fermentation stage. Cassava starch processing wastewater fermentation effluent consisted of acetic acid, butyric acid and propionic acid. The effluent from fermentation stage was used as feedstock to generate hydrogen production by microbial electrolysis cell (MECs) at an applied voltage of 0.6 V in second stage with additional 657 mL H2/g-COD was produced. Energy efficiencies based on electricity needed for the MEC were 330 % with COD removals of 95 %. The overall hydrogen yield was 800-900 mL H2/g-COD. Microbial community analysis of electrohydrogenesis by DGGE shows that exoelectrogens belong to Acidiphilium sp., Geobacter sulfurreducens and Thermincola sp. were dominated at anode. These results show two-stage thermophilic fermentation, and electrohydrogenesis process improved hydrogen production performance with high hydrogen yields, high gas production rates and high COD removal efficiency.

Keywords: cassava starch processing wastewater, biohydrogen, thermophilic fermentation, microbial electrolysis cell

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1171 Insights into the Annotated Genome Sequence of Defluviitoga tunisiensis L3 Isolated from a Thermophilic Rural Biogas Producing Plant

Authors: Irena Maus, Katharina Gabriella Cibis, Andreas Bremges, Yvonne Stolze, Geizecler Tomazetto, Daniel Wibberg, Helmut König, Alfred Pühler, Andreas Schlüter


Within the agricultural sector, the production of biogas from organic substrates represents an economically attractive technology to generate bioenergy. Complex consortia of microorganisms are responsible for biomass decomposition and biogas production. Recently, species belonging to the phylum Thermotogae were detected in thermophilic biogas-production plants utilizing renewable primary products for biomethanation. To analyze adaptive genome features of representative Thermotogae strains, Defluviitoga tunisiensis L3 was isolated from a rural thermophilic biogas plant (54°C) and completely sequenced on an Illumina MiSeq system. Sequencing and assembly of the D. tunisiensis L3 genome yielded a circular chromosome with a size of 2,053,097 bp and a mean GC content of 31.38%. Functional annotation of the complete genome sequence revealed that the thermophilic strain L3 encodes several genes predicted to facilitate growth of this microorganism on arabinose, galactose, maltose, mannose, fructose, raffinose, ribose, cellobiose, lactose, xylose, xylan, lactate and mannitol. Acetate, hydrogen (H2) and carbon dioxide (CO2) are supposed to be end products of the fermentation process. The latter gene products are metabolites for methanogenic archaea, the key players in the final step of the anaerobic digestion process. To determine the degree of relatedness of dominant biogas community members within selected digester systems to D. tunisiensis L3, metagenome sequences from corresponding communities were mapped on the L3 genome. These fragment recruitments revealed that metagenome reads originating from a thermophilic biogas plant covered 95% of D. tunisiensis L3 genome sequence. In conclusion, availability of the D. tunisiensis L3 genome sequence and insights into its metabolic capabilities provide the basis for biotechnological exploitation of genome features involved in thermophilic fermentation processes utilizing renewable primary products.

Keywords: genome sequence, thermophilic biogas plant, Thermotogae, Defluviitoga tunisiensis

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1170 Clostridium thermocellum DBT-IOC-C19, A Potential CBP Isolate for Ethanol Production

Authors: Nisha Singh, Munish Puri, Collin Barrow, Deepak Tuli, Anshu S. Mathur


The biological conversion of lignocellulosic biomass to ethanol is a promising strategy to solve the present global crisis of exhausting fossil fuels. The existing bioethanol production technologies have cost constraints due to the involvement of mandate pretreatment and extensive enzyme production steps. A unique process configuration known as consolidated bioprocessing (CBP) is believed to be a potential cost-effective process due to its efficient integration of enzyme production, saccharification, and fermentation into one step. Due to several favorable reasons like single step conversion, no need of adding exogenous enzymes and facilitated product recovery, CBP has gained the attention of researchers worldwide. However, there are several technical and economic barriers which need to be overcome for making consolidated bioprocessing a commercially viable process. Finding a natural candidate CBP organism is critically important and thermophilic anaerobes are preferred microorganisms. The thermophilic anaerobes that can represent CBP mainly belong to genus Clostridium, Caldicellulosiruptor, Thermoanaerobacter, Thermoanaero bacterium, and Geobacillus etc. Amongst them, Clostridium thermocellum has received increased attention as a high utility CBP candidate due to its highest growth rate on crystalline cellulose, the presence of highly efficient cellulosome system and ability to produce ethanol directly from cellulose. Recently with the availability of genetic and molecular tools aiding the metabolic engineering of Clostridium thermocellum have further facilitated the viability of commercial CBP process. With this view, we have specifically screened cellulolytic and xylanolytic thermophilic anaerobic ethanol producing bacteria, from unexplored hot spring/s in India. One of the isolates is a potential CBP organism identified as a new strain of Clostridium thermocellum. This strain has shown superior avicel and xylan degradation under unoptimized conditions compared to reported wild type strains of Clostridium thermocellum and produced more than 50 mM ethanol in 72 hours from 1 % avicel at 60°C. Besides, this strain shows good ethanol tolerance and growth on both hexose and pentose sugars. Hence, with further optimization this new strain could be developed as a potential CBP microbe.

Keywords: Clostridium thermocellum, consolidated bioprocessing, ethanol, thermophilic anaerobes

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1169 Thermophilic Anaerobic Granular Membrane Distillation Bioreactor for Wastewater Reuse

Authors: Duong Cong Chinh, Shiao-Shing Chen, Le Quang Huy


Membrane distillation (MD) is actually claimed to be a cost-effective separation process when waste heat, alternative energy sources, or wastewater are used. To the best of our knowledge, this is the first study that a thermophilic anaerobic granular bioreactor is integrated with membrane distillation (ThAnMDB) was investigated. In this study, the laboratory scale anaerobic bioreactor (1.2 litter) was set-up. The bioreactor was maintained at temperature 55 ± 2°C, hydraulic retention time = 0.5 days, organic loading rates of 7 and 10 kg chemical oxygen demand (COD) m³/day. Side-stream direct contact membrane distillation with the polytetrafluoroethylene membrane area was 150 cm². The temperature of the distillate was kept at 25°C. Results show that distillate flux was 19.6 LMH (Liters per square meter per hour) on the first day and gradually decreased to 6.9 LMH after 10 days, and the membrane was not wet. Notably, by directly using the heat from the thermophilic anaerobic for MD separation process, all distilled water from wastewater was reuse as fresh water (electrical conductivity < 120 µs/cm). The ThAnMDB system showed its high pollutant removal performance: chemical oxygen demand (COD) from 99.6 to 99.9%, NH₄⁺ from 60 to 95%, and PO₄³⁻ complete removal. In addition, methane yield was from 0.28 to 0.34 lit CH₄/gram COD removal (80 – 97% of the theoretical) demonstrated that the ThAnMDB system was quite stable. The achievement of the ThAnMDB is not only in removing pollutants and reusing wastewater but also in absolutely unnecessarily adding alkaline to the anaerobic bioreactor system.

Keywords: high rate anaerobic digestion, membrane distillation, thermophilic anaerobic, wastewater reuse

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1168 Evaluation of Antimicrobial Activity of Different Dithiolethiones

Authors: Zehour Rahmani, Messouda Dekmouche, Mohamed Hadjadj, Mokhtar Saidi


In the last decades of the nineteenth century, the study of disease – causing microorganisms became concentrated on bacteria and largely institutionalized. In earlier years, the scientists interested in bacteria had originally been chemists like Pasteur, physicists like Tyndall, or botanists like Cohn and ward. For this reason, the objective of this research was to evaluate the potential of some dithiolethiones on standard microorganism strains as well as multi-drug resistant bacteria, which were isolated from hospitals. Recent studies have demonstrated, that several dithiolethione compounds, particularly (3H-1,2-dithiole-3-thione), exhibit the biological activities against several bacteria.

Keywords: bacteria, dithiolethiones, microorganism, potential

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1167 Investigations of Protein Aggregation Using Sequence and Structure Based Features

Authors: M. Michael Gromiha, A. Mary Thangakani, Sandeep Kumar, D. Velmurugan


The main cause of several neurodegenerative diseases such as Alzhemier, Parkinson, and spongiform encephalopathies is formation of amyloid fibrils and plaques in proteins. We have analyzed different sets of proteins and peptides to understand the influence of sequence-based features on protein aggregation process. The comparison of 373 pairs of homologous mesophilic and thermophilic proteins showed that aggregation-prone regions (APRs) are present in both. But, the thermophilic protein monomers show greater ability to ‘stow away’ the APRs in their hydrophobic cores and protect them from solvent exposure. The comparison of amyloid forming and amorphous b-aggregating hexapeptides suggested distinct preferences for specific residues at the six positions as well as all possible combinations of nine residue pairs. The compositions of residues at different positions and residue pairs have been converted into energy potentials and utilized for distinguishing between amyloid forming and amorphous b-aggregating peptides. Our method could correctly identify the amyloid forming peptides at an accuracy of 95-100% in different datasets of peptides.

Keywords: aggregation, amyloids, thermophilic proteins, amino acid residues, machine learning techniques

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1166 Production of Poly-β-Hydroxybutyrate (PHB) by a Thermophilic Strain of Bacillus and Pseudomonas Species

Authors: Patience Orobosa Olajide


Five hydrocarbon degrading bacterial strains isolated from contaminated environment were investigated with respect to polyhydroxybutyrate (PHB) biosynthesis. Screening for bioplastic production was done on assay mineral salts agar medium containing 0.2% poly (3-hydroxybutyrate) as the sole carbon source. Two of the test bacteria were positive for PHB biosynthesis and were identified based on gram staining, biochemical tests, 16S rRNA gene sequence analysis as Pseudomonas aeruginosa and Bacillus licheniformis which grew at 37 and up to 65 °C respectively, thus suggesting the later to be thermotolerant. In this study, the effects of different carbon and nitrogen sources on PHB production in these strains were investigated. Maximum PHB production was obtained in 48 hr for the two strains and amounted to yields of 72.86 and 62.22 percentages for Bacillus licheniformis and Pseudomonas aeruginosa respectively. In these strains, glycine was the most efficient carbon sources for the production of PHB compared with other carbon (glucose, lactose, sucrose, Arabinose) and nitrogen (L- glycine, L-cysteine, DL-Tryptophan, and Potassium Nitrate) sources. The screening of microbial strains for industrial PHB production should be based on several factors including the cell’s capability to mineralize an inexpensive substrate, rate of growth and the extent of polymer accumulation.

Keywords: bacteria, poly-3-hydroxybutyrate (PHB), hydrocarbon, thermotolerant

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1165 Effects of Bacteria on Levels of AFM1 in Phosphate Buffer at Different Level of Energy Source

Authors: Ali M. Elgerbi, Obied A. Alwan, Al-Taher O. Alzwei, Abdurrahim A. Elouzi


The binding of AFM1 to bacteria in phosphate buffer solution depended on many factors such as: availability of energy, incubation period, species and strain of bacteria. Increase in concentration of sugar showed higher removal of AFM1 and faster than in phosphate buffer alone. With 1.0% glucose lactic acid bacteria and bifidobacteria showed toxin removal ranging from 7.7 to 39.7% whereas with 10.0% glucose the percentage removal was 21.8 to 45.4% at 96 hours of incubation.

Keywords: aflatoxin M1, lactic acid bacteria, bifidobacteria , binding, phosphate buffer

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1164 Anaerobic Digestion of Green Wastes at Different Solids Concentrations and Temperatures to Enhance Methane Generation

Authors: A. Bayat, R. Bello-Mendoza, D. G. Wareham


Two major categories of green waste are fruit and vegetable (FV) waste and garden and yard (GY) waste. Although, anaerobic digestions (AD) is able to manage FV waste; there is less confidence in the conditions for AD to handle GY wastes (grass, leaves, trees and bush trimmings); mainly because GY contains lignin and other recalcitrant organics. GY in the dry state (TS ≥ 15 %) can be digested at mesophilic temperatures; however, little methane data has been reported under thermophilic conditions, where conceivably better methane yields could be achieved. In addition, it is suspected that at lower solids concentrations, the methane yield could be increased. As such, the aim of this research is to find the temperature and solids concentration conditions that produce the most methane; under two different temperature regimes (mesophilic, thermophilic) and three solids states (i.e. 'dry', 'semi-dry' and 'wet'). Twenty liters of GY waste was collected from a public park located in the northern district in Tehran. The clippings consisted of freshly cut grass as well as dry branches and leaves. The GY waste was chopped before being fed into a mechanical blender that reduced it to a paste-like consistency. An initial TS concentration of approximately 38 % was achieved. Four hundred mL of anaerobic inoculum (average total solids (TS) concentration of 2.03 ± 0.131 % of which 73.4% were volatile solid (VS), soluble chemical oxygen demand (sCOD) of 4.59 ± 0.3 g/L) was mixed with the GY waste substrate paste (along with distilled water) to achieve a TS content of approximately 20 %. For comparative purposes, approximately 20 liters of FV waste was ground in the same manner as the GY waste. Since FV waste has a much higher natural water content than GY, it was dewatered to obtain a starting TS concentration in the dry solid-state range (TS ≥ 15 %). Three samples were dewatered to an average starting TS concentration of 32.71 %. The inoculum was added (along with distilled water) to dilute the initial FV TS concentrations down to semi-dry conditions (10-15 %) and wet conditions (below 10 %). Twelve 1-L batch bioreactors were loaded simultaneously with either GY or FV waste at TS solid concentrations ranging from 3.85 ± 1.22 % to 20.11 ± 1.23 %. The reactors were sealed and were operated for 30 days while being immersed in water baths to maintain a constant temperature of 37 ± 0.5 °C (mesophilic) or 55 ± 0.5 °C (thermophilic). A maximum methane yield of 115.42 (L methane/ kg VS added) was obtained for the GY thermophilic-wet AD combination. Methane yield was enhanced by 240 % compared to the GY waste mesophilic-dry condition. The results confirm that high temperature regimes and small solids concentrations are conditions that enhance methane yield from GY waste. A similar trend was observed for the anaerobic digestion of FV waste. Furthermore, a maximum value of VS (53 %) and sCOD (84 %) reduction was achieved during the AD of GY waste under the thermophilic-wet condition.

Keywords: anaerobic digestion, thermophilic, mesophilic, total solids concentration

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1163 Bacteria Removal from Wastewater by Electrocoagulation Process

Authors: Boudjema Nouara, Mameri Nabil


Bacteria have played an important role in water contamination as a consequence of organic pollution. In this study, an electrocoagulation process was adopted to remove fecal contamination and pathogenic bacteria from waste water. The effect of anode/cathodes materials as well as operating conditions for bacteria removal from water, such as current intensity and initial pH and temperature. The results indicated that the complete removal was achevied when using aluminium anode as anode at current intensity of 3A, initial pH of 7-8 and electrolysis time of 30 minutes. This process showed a bactericidal effect of 95 to 99% for the total and fecal coliforms and 99% to 100% for Eschercichia coli and fecal Streptococci. A decrease of 72% was recorded for sulphite-reducing Clostridia. Thus, this process has the potential to be one the options for treatment where high amount of bacteria in wastewater river.

Keywords: bacteria, el Harrach river, electrocoagulation, wastewater, treatment

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1162 Anaerobic Digestion of Spent Wash through Biomass Development for Obtaining Biogas

Authors: Sachin B. Patil, Narendra M. Kanhe


A typical cane molasses based distillery generates 15 L of waste water per liter of alcohol production. Distillery waste with COD of over 1,00,000 mg/l and BOD of over 30,000 mg/l ranks high amongst the pollutants produced by industries both in magnitude and strength. Treatment and safe disposal of this waste is a challenging task since long. The high strength of waste water renders aerobic treatment very expensive and physico-chemical processes have met with little success. Thermophilic anaerobic treatment of distillery waste may provide high degree of treatment and better recovery of biogas. It may prove more feasible in most part of tropical country like India, where temperature is suitable for thermophilic micro-organisms. Researchers have reviled that, at thermophilic conditions due to increased destruction rate of organic matter and pathogens, higher digestion rate can be achieved. Literature review reveals that the variety of anaerobic reactors including anaerobic lagoon, conventional digester, anaerobic filter, two staged fixed film reactors, sludge bed and granular bed reactors have been studied, but little attempts have been made to evaluate the usefulness of thermophilic anaerobic treatment for treating distillery waste. The present study has been carried out, to study feasibility of thermophilic anaerobic digestion to facilitate the design of full scale reactor. A pilot scale anaerobic fixed film fixed bed reactor (AFFFB) of capacity 25m3 was designed, fabricated, installed and commissioned for thermophilic (55-65°C) anaerobic digestion at a constant pH of 6.5-7.5, because these temperature and pH ranges are considered to be optimum for biogas recovery from distillery wastewater. In these conditions, working of the reactor was studied, for different hydraulic retention times (HRT) (0.25days to 12days) and variable organic loading rates (361.46 to 7.96 Kg COD/m3d). The parameters such as flow rate and temperature, various chemical parameters such as pH, chemical oxygen demands (COD), biogas quantity, and biogas composition were regularly monitored. It was observed that, with the increase in OLR, the biogas production was increased, but the specific biogas yield decreased. Similarly, with the increase in HRT, the biogas production got decrease, but the specific biogas yield was increased. This may also be due to the predominant activity of acid producers to methane producers at the higher substrate loading rates. From the present investigation, it can be concluded that for thermophilic conditions the highest COD removal percentage was obtained at an HRT of 08 days, thereafter it tends to decrease from 8 to 12 days HRT. There is a little difference between COD removal efficiency of 8 days HRT (74.03%) and 5 day HRT (78.06%), therefore it would not be feasible to increase the reactor size by 1.5 times for mere 4 percent more efficiency. Hence, 5 days HRT is considered to be optimum, at which the biogas yield was 98 m3/day and specific biogas yield was 0.385 CH4 m3/Kg CODr.

Keywords: spent wash, anaerobic digestion, biomass, biogas

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1161 Isolation and Characterization of Indigenous Rhizosphere Bacteria Producing Gibberellin Acid from Local Soybeans in Three Different Areas of South Sulawesi

Authors: Asmiaty Sahur, Ambo Ala, Baharuddin Patanjengi, Elkawakib Syam'un


This study aimed to isolate and characterize the indigenous Rhizosphere bacteria producing Gibberellin Acid as plant growth isolated from local soybean of three different areas in South Sulawesi, Indonesia. Several soil samples of soybean plants were collected from the Rhizosphere of local soybeans in three different areas of South Sulawesi such as Soppeng, Bone and Takalar. There were 56 isolates of bacteria were isolated and grouped into gram-positive bacteria and gram negative bacteria .There are 35 isolates produce a thick slime or slimy when cultured on media Natrium Broth and the remaining of those produced spores. The results showed that of potential bacterial isolated produced Gibberellin Acid in high concentration. The best isolate of Rhizosphere bacteria for the production of Gibberellin Acid is with concentration 2%. There are 4 isolates that had higher concentration are AKB 19 (4.67 mg/ml) followed by RKS 17 (3.80 mg/ml), RKS 25 (3.70 mg / ml) and RKS 24 (3.29 mg/ml) respectively.

Keywords: rhizosphere, bacteria, gibberellin acid, soybeans

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1160 The Effect of Bacteria on Mercury's Biological Removal

Authors: Nastaran Soltani


Heavy metals such as Mercury are toxic elements that enter the environment through different ways and endanger the environment, plants, animals, and humans’ health. Microbial activities reduce the amount of heavy metals. Therefore, an effective mechanism to eliminate heavy metals in the nature and factory slops, is using bacteria living in polluted areas. Karun River in Khuzestan Province in Iran has been always polluted by heavy metals as it is located among different industries in the region. This study was performed based on the data from sampling water and sediments of four stations across the river during the four seasons of a year. The isolation of resistant bacteria was performed through enrichment and direct cultivation in a solid medium containing mercury. Various bacteria such as Pseudomonas sp., Serratia Marcescens, and E.coli were identified as mercury-resistant bacteria. The power of these bacteria to remove mercury varied from 28% to 86%, with strongest power belonging to Pseudomonas sp. isolated in spring making a good candidate to be used for mercury biological removal from factory slops.

Keywords: bacteria, Karun River, mercury, biological removal, mercury-resistant

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1159 Examining the Presence of Heterotrophic Aerobic Bacteria (HAB), and Sulphate Reducing Bacteria (SRB) in Some Types of Water from the City of Tripoli, Libya

Authors: Abdulsalam. I. Rafida, Marwa. F. Elalem, Hasna. E. Alemam


This study aimed at testing the various types of water in some areas of the city of Tripoli, Libya for the presence of Heterotrophic Aerobic Bacteria (HAB), and anaerobic Sulphate Reducing Bacteria (SRB). The water samples under investigation included rainwater accumulating on the ground, sewage water (from the city sewage treatment station, sulphate water from natural therapy swimming sites), and sea water (i.e. sea water exposed to pollution by untreated sewage water, and unpolluted sea water from specific locations). A total of 20 samples have been collected distributed as follows: rain water (8 samples), sewage water (6 samples), and sea water (6 samples). An up-to-date method for estimation has been used featuring readymade solutions i.e. (BARTTM test for HAB and BARTTM test for SRB). However, with the exception of one rain water sample, the results have indicated that the target bacteria have been present in all samples. Regarding HAB bacteria the samples have shown a maximum average of 7.0 x 106 cfu/ml featuring sewage and rain water and a minimum average of 1.8 x 104 cuf/ml featuring unpolluted sea water collected from a specific location. As for SRB bacteria; a maximum average of 7.0 x 105 cfu/ml has been shown by sewage and rain water and a minimum average of 1.8 x 104 cfu/ml by sewage and sea water. The above results highlight the relationship between pollution and the presence of bacteria in water particularly water collected from specific locations, and also the presence of bacteria as the result of the use of water provided that a suitable environment exists for its growth.

Keywords: heterotrophic aerobic bacteria (HAB), sulphate reducing bacteria (SRB), water, environmental sciences

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1158 Isolation, Screening and Identification of Frog Cutaneous Bacteria for Anti-Batrachochytrium dendrobatidis Activity

Authors: Adria Rae Abigail R. Eda, Arvin C. Diesmos, Vance T. Vredenburg, Merab A. Chan


Mitigating strategies using symbiotic cutaneous bacteria is one of the major concerns in the conservation of amphibian population. Batrachochytrium dendrobatidis is the causative agent of chytridiomycosis associated with mass mortality and amphibian extinctions worldwide. In the Philippines, there is a lack of study on the cutaneous bacteria of Philippine amphibians that may have beneficial effects to ward off the deadly fungal infection. In this study, cutaneous bacteria from frogs were isolated and examined for anti-B. dendrobatidis activity. Eight species of frogs were collected at Mt. Palay-palay Mataas na Gulod National Park in Cavite, a site positive for the presence of B. dendrobatidis. Bacteria were isolated from the skin of frogs by swabbing the surfaces of the body and inoculated in Reasoner´s 2A (R2A) agar. Isolated bacteria were tested for potential inhibitory properties against B. dendrobatidis through zoospore inhibition assay. Results showed that frog cutaneous bacteria significantly inhibited the growth of B. dendrobatidis in vitro. By means of 16S rRNA gene primers, the anti-B. dendrobatidis bacteria were identified to be Enterobacter sp., Alcaligenes faecalis and Pseudomonas sp. Cutaneous bacteria namely Enterobacter sp. (isolates PLd33 and PCv4) and Pseudomonas (isolate PLd31) remarkably cleared the growth of B. dendrobatidis zoospore in 1% tryptone agar. Therefore, frog cutaneous bacteria inhibited B. dendrobatidis in vitro and could possibly contribute to the immunity and defense of frogs against the lethal chytridiomycosis.

Keywords: Batrachochytrium dendrobatidis, cutaneous bacteria, frogs, zoospore inhibition assay

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1157 Lactobacillus sp. Isolates Slaughterhouse Waste as Probiotics for Broilers

Authors: Nourmalita Safitri Ningsih, Ridwan, Iqri Puspa Yunanda


The aim of this study was to utilize the waste from slaughterhouses for chicken feed ingredients is probiotic. Livestock waste produced by livestock activities such as feces, urine, food remains, as well as water from livestock and cage cleaning. The process starts with the isolation of bacteria. Rumen fluid is taken at Slaughterhouse Giwangan, Yogyakarta. Isolation of Lactobacillus ruminus is done by using de Mann Rogosa Sharpe (MRS) medium. In the sample showed a rod-shaped bacteria are streaked onto an agar plates. After it was incubated at 37ºC for 48 hours, after which it is observed. The observation of these lactic acid bacteria it will show a clear zone at about the colony. These bacterial colonies are white, round, small, shiny on the agar plate mikroenkapsul In the manufacturing process carried out by the method of freeze dried using skim milk in addition capsulated material. Then the results of these capsulated bacteria are mixed with feed for livestock. The results from the mixing of capsulated bacteria in feed are to increase the quality of animal feed so as to provide a good effect on livestock. Scanning electron microscope testing we have done show the results of bacteria have been shrouded in skim milk. It can protect the bacteria so it is more durable in use. The observation of the bacteria showed a sheath on Lactobacillus sp. Preservation of bacteria in this way makes the bacteria more durable for use. As well as skim milk can protect bacteria that are resistant to the outside environment. Results of probiotics in chicken feed showed significant weight gain in chickens. Calculation Anova (P <0.005) shows the average chicken given probiotics her weight increased.

Keywords: chicken, probiotics, waste, Lactobacillus sp, bacteria

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1156 Study on the Heavy Oil Degradation Performance and Kinetics of Immobilized Bacteria on Modified Zeolite

Authors: Xiao L Dai, Wen X Wei, Shuo Wang, Jia B Li, Yan Wei


Heavy oil pollution generated from both natural and anthropogenic sources could cause significant damages to the ecological environment, due to the toxicity of some of its constituents. Nowadays, microbial remediation is becoming a promising technology to treat oil pollution owing to its low cost and prevention of secondary pollution; microorganisms are key players in the process. Compared to the free microorganisms, immobilized microorganisms possess several advantages, including high metabolic activity rates, strong resistance to toxic chemicals and natural competition with the indigenous microorganisms, and effective resistance to washing away (in open water system). Many immobilized microorganisms have been successfully used for bioremediation of heavy oil pollution. Considering the broad choices, low cost, simple process, large specific surface area and less impact on microbial activity, modified zeolite were selected as a bio-carrier for bacteria immobilization. Three strains of heavy oil-degrading bacteria Bacillus sp. DL-13, Brevibacillus sp. DL-1 and Acinetobacter sp. DL-34 were immobilized on the modified zeolite under mild conditions, and the bacterial load (bacteria /modified zeolite) was 1.12 mg/g, 1.11 mg/g, and 1.13 mg/g, respectively. SEM results showed that the bacteria mainly adsorbed on the surface or punctured in the void of modified zeolite. The heavy oil degradation efficiency of immobilized bacteria was 62.96%, higher than that of the free bacteria (59.83%). The heavy oil degradation process of immobilized bacteria accords with the first-order reaction equation, and the reaction rate constant is 0.1483 d⁻¹, which was significantly higher than the free bacteria (0.1123 d⁻¹), suggesting that the immobilized bacteria can rapidly start up the heavy oil degradation and has a high activity of heavy oil degradation. The results suggested that immobilized bacteria are promising technology for bioremediation of oil pollution.

Keywords: heavy oil pollution, microbial remediation, modified zeolite, immobilized bacteria

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1155 Biodiversity of Plants Rhizosphere and Rhizoplane Bacteria in the Presence of Petroleum Hydrocarbons

Authors: Togzhan D. Mukasheva, Anel A. Omirbekova, Raikhan S. Sydykbekova, Ramza Zh. Berzhanova, Lyudmila V. Ignatova


Following plants-barley (Hordeum sativum), alfalfa (Medicago sativa), grass mixture (red fescue-75%, long-term ryegrass - 20% Kentucky bluegrass - 10%), oilseed rape (Brassica napus biennis), resistant to growth in the contaminated soil with oil content of 15.8 g / kg 25.9 g / kg soil were used. Analysis of the population showed that the oil pollution reduces the number of bacteria in the rhizosphere and rhizoplane of plants and enhances the amount of spore-forming bacteria and saprotrophic micromycetes. It was shown that regardless of the plant, dominance of Pseudomonas and Bacillus genera bacteria was typical for the rhizosphere and rhizoplane of plants. The frequency of bacteria of these genera was more than 60%. Oil pollution changes the ratio of occurrence of various types of bacteria in the rhizosphere and rhizoplane of plants. Besides the Pseudomonas and Bacillus genera, in the presence of hydrocarbons in the root zone of plants dominant and most typical were the representatives of the Mycobacterium and Rhodococcus genera. Together the number was between 62% to 72%.

Keywords: pollution, root system, micromycetes, identification

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1154 Phylogenetic Characterization of Atrazine-Degrading Bacteria Isolated from Agricultural Soil in Eastern Thailand

Authors: Sawangjit Sopid


In this study sugarcane field soils with a long history of atrazine application in Chachoengsao and Chonburi provinces have been explored for their potential of atrazine biodegradation. For the atrazine degrading bacteria isolation, the soils used in this study named ACS and ACB were inoculated in MS-medium containing atrazine. Six short rod and gram-negative bacterial isolates, which were able to use this herbicide as a sole source of nitrogen, were isolated and named as ACS1, ACB1, ACB3, ACB4, ACB5 and ACB6. From the 16S rDNA nucleotide sequence analysis, the isolated bacteria ACS1 and ACB4 were identified as Rhizobium sp. with 89.1-98.7% nucleotide identity, ACB1 and ACB5 were identified as Stenotrophomonas sp. with 91.0-92.8% nucleotide identity, whereas ACB3 and ACB6 were Klebsiella sp. with 97.4-97.8% nucleotide identity.

Keywords: atrazine-degrading bacteria, bioremediation, Thai isolates, bacteria

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1153 Fungal Cellulase/Xylanase Complex and Their Industrial Applications

Authors: L. Kutateldze, T. Urushadze, R. Khvedelidze, N. Zakariashvili, I. Khokhashvili, T. Sadunishvili


Microbial cellulase/xylanase have shown their potential application in various industries including pulp and paper, textile, laundry, biofuel production, food and feed industry, brewing, and agriculture. Extremophilic micromycetes and their enzymes that are resistant to critical values of temperature and pH, and retaining enzyme activity for a long time are of great industrial interest. Among strains of microscopic fungi from the collection of S. Durmishidze Institute of Biochemistry and Biotechnology, strains isolated from different ecological niches of Southern Caucasus-active producers of cellulase/xylanase have been selected by means of screening under deep cultivation conditions. Extremophilic micromycetes and their enzymes that are resistant to critical values of temperature and pH, and retaining enzyme activity for a long time are of great industrial interest. Among strains of microscopic fungi from the collection of S. Durmishidze Institute of Biochemistry and Biotechnology, strains isolated from different ecological niches of Southern Caucasus-active producers of cellulase/xylanase have been selected by means of screening under deep cultivation conditions. Representatives of the genera Aspergillus, Penicillium and Trichoderma are outstanding by relatively high activities of these enzymes. Among the producers were revealed thermophilic strains, representatives of the genus Aspergillus-Aspergillus terreus, Aspergillus versicolor, Aspergillus wentii, also strains of Sporotrichum pulverulentum and Chaetomium thermophile. As a result of optimization of cultivation media and conditions, activities of enzymes produced by the strains have been increased by 4 -189 %. Two strains, active producers of cellulase/xylanase – Penicillium canescence E2 (mesophile) and Aspergillus versicolor Z17 (thermophile) were chosen for further studies. Cellulase/xylanase enzyme preparations from two different genera of microscopic fungi Penicillium canescence E2 and Aspergillus versicolor Z 17 were obtained with activities 220 U/g /1200 U/g and 125 U/g /940 U/g, correspondingly. Main technical characteristics were as follows: the highest enzyme activities were obtained for mesophilic strain Penicillium canescence E2 at 45-500C, while almost the same enzyme activities were fixed for the thermophilic strain Aspergillus versicolor Z 17 at temperature 60-65°C, exceeding the temperature optimum of the mesophile by 150C. Optimum pH of action of the studied cellulase/xylanases from mesophileic and thermophilic strains were similar and equaled to 4.5-5.0 It has been shown that cellulase/xylanase technical preparations from selected strains of Penicillium canescence E2 and Aspergillus versicolor Z17 hydrolyzed cellulose of untreated wheat straw to reducible sugars by 46-52%, and to glucose by 22-27%. However the thermophilic enzyme preparations from the thermophilic A.versicolor strains conducted the process at 600C higher by 100C as compared to mesophlic analogue. Rate of hydrolyses of the pretreated substrate by the same enzyme preparations to reducible sugars and glucose conducted at optimum for their action 60 and 500C was 52-61% and 29-33%, correspondingly. Thus, maximum yield of glucose and reducible sugars form untreated and pretreated wheat straw was achieved at higher temperature (600C) by enzyme preparations from thermophilic strain, which gives advantage for their industrial application.

Keywords: cellulase/xylanase, cellulose hydrolysis, microscopic fungi, thermophilic strain

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1152 Characterization of Bacteria by a Nondestructive Sample Preparation Method in a TEM System

Authors: J. Shiue, I. H. Chen, S. W. Y. Chiu, Y. L. Wang


In this work, we present a nondestructive method to characterize bacteria in a TEM system. Unlike the conventional TEM specimen preparation method, which needs to thin the specimen in a destructive way, or spread the samples on a tiny millimeter sized carbon grid, our method is easy to operate without the need of sample pretreatment. With a specially designed transparent chip that allows the electron beam to pass through, and a custom made chip holder to fit into a standard TEM sample holder, the bacteria specimen can be easily prepared on the chip without any pretreatment, and then be observed under TEM. The centimeter-sized chip is covered with Au nanoparticles in the surface as the markers which allow the bacteria to be observed easily on the chip. We demonstrate the success of our method by using E. coli as an example, and show that high-resolution TEM images of E. coli can be obtained with the method presented. Some E. coli morphology characteristics imaged using this method are also presented.

Keywords: bacteria, chip, nanoparticles, TEM

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1151 Use of Lactic Strains Isolated from Algerian Ewe's Milk in the Manufacture of a Natural Yogurt

Authors: Chougrani Fadela, Cheriguene Abderrahim


Fifty three strains of thermophilic and mesophilic lactic acid bacteria were isolated from the ewe’s milk. Identification reveals the presence of nineteen strains (36%) of Lactobacillus sp., seventeen strains (32%) of Lactococcus sp., nine strains (17%) of Streptococcus thermophilus and eight strains (15%) of Leuconostoc sp. The strains were characterized for their technological properties. A high diversity of properties among the studied strains was demonstrated. On the basis of technological characteristics, two strains (Lactobacillus bulgaricus and Streptococcus thermophilus) were screened with respect to their acid and flavour production for the preparation of a natural yogurt and compared to a commercial starter cultures. Sensorial analyses revealed that the product manufactured on the basis of the isolated strains have a cohesiveness and adhesiveness corresponding to standard products. The pH and the acidity recorded are also within accepted levels during all the period of conservation.

Keywords: Lactobacillus bulgaricus, Streptococcus thermophilus, yoghurt, cohesiveness, adhesiveness, Algerian ewe’s milk

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1150 On a Negative Relation between Bacterial Taxis and Turing Pattern Formation

Authors: A. Elragig, S. Townley, H. Dreiwi


In this paper we introduce a bacteria-leukocyte model with bacteria chemotaxsis. We assume that bacteria develop a tactic defense mechanism as a response to Leukocyte phagocytosis. We explore the effect of this tactic motion on Turing space in two parameter spaces. A fine tuning of bacterial chemotaxis shows a significant effect on developing a non-uniform steady state.

Keywords: chemotaxis-diffusion driven instability, bacterial chemotaxis, mathematical biology, ecology

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