Search results for: bacteria and Fungi.

Authors: Abimbola M. Enitan, John O. Odiyo, Feroz M. Swalaha

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The study of microbial ecology and their function in anaerobic digestion processes are essential to control the biological processes. This is to know the symbiotic relationship between the microorganisms that are involved in the conversion of complex organic matter in the industrial wastewater to simple molecules. In this study, diversity and quantity of bacterial community in the granular sludge taken from the different compartments of a full-scale upflow anaerobic sludge blanket (UASB) reactor treating brewery wastewater was investigated using polymerase chain reaction (PCR) and real-time quantitative PCR (qPCR). The phylogenetic analysis showed three major eubacteria phyla that belong to Proteobacteria, Firmicutes and Chloroflexi in the full-scale UASB reactor, with different groups populating different compartment. The result of qPCR assay showed high amount of eubacteria with increase in concentration along the reactor’s compartment. This study extends our understanding on the diverse, topological distribution and shifts in concentration of microbial communities in the different compartments of a full-scale UASB reactor treating brewery wastewater. The colonization and the trophic interactions among these microbial populations in reducing and transforming complex organic matter within the UASB reactors were established.

Keywords: Bacteria, brewery wastewater, real-time quantitative PCR, UASB reactor.

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Authors: Ime R. Udotong, Ofonime U. M. John, Justina I. R. Udotong

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The microbiological and physicochemical characteristics of wetland soils in Eket Local Government Area were studied between May 2001 and June 2003. Total heterotrophic bacterial counts (THBC), total fungal counts (TFC), and total actinomycetes counts (TAC) were determined from soil samples taken from four locations at two depths in the wet and dry seasons. Microbial isolates were characterized and identified. Particle size and chemical parameters were also determined using standard methods. THBC ranged from 5.2 (+0.17) x106 to 1.7 (+0.18) x107 cfu/g and from 2.4 (+0.02) x106 to 1.4 (+0.04) x107cfu/g in the wet and dry seasons, respectively. TFC ranged from 1.8 (+0.03) x106 to 6.6 (+ 0.18) x106 cfu/g and from 1.0 (+0.04) x106 to 4.2 (+ 0.01) x106 cfu/g in the wet and dry seasons, respectively .TAC ranged from 1.2 (+0.53) x106 to 6.0 (+0.05) x106 cfu/g and from 0.6 (+0.01) x106 to 3.2 (+ 0.12) x106 cfu/g in the wet and dry season, respectively. Acinetobacter, Alcaligenes, Arthrobacter, Bacillus, Beijerinckja, Enterobacter, Micrococcus, Flavobacterium, Serratia, Enterococcus, and Pseudomonas species were predominant bacteria while Aspergillus, Fusarium, Mucor, Penicillium, and Rhizopus were the dominant fungal genera isolated. Streptomyces and Norcadia were the actinomycetes genera isolated. The particle size analysis showed high sand fraction but low silt and clay. The pH and % organic matter were generally acidic and low, respectively at all locations. Calcium dominated the exchangeable bases with low electrical conductivity and micronutrients. These results provide the baseline data of Eket wetland soils for its management for sustainable agriculture.

Keywords: Wetland soils, Microbial counts, physicochemicalcharacteristics, Sustainable agriculture.

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Authors: Rupinder Kaur, Parmjit S. Panesar, Ram S. Singh

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Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050 was observed to be the maximum extracellular enzyme producer.

Keywords: β-galactosidase, fungus, yeast, whey.

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Authors: Z. Honarvar, M. Farhoodi, M. R. Khani, S. Shojaee-Aliabadi

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The purpose of the present study was to evaluate carboxymethyl cellulose (CMC) coated polypropylene (PP) films containing Zataria multiflora (ZEO) essential oils (4%) as an antimicrobial packaging for chicken breast stored at 4 °C. To increase PP film hydrophilicity, it was treated by atmospheric cold plasma prior to coating by CMC. Then, different films including PP, PP/CMC, PP/CMC containing 4% of ZEO were used for the chicken meat packaging in vapor phase. Total viable count and pseudomonads population and oxidative (TBA) changes of the chicken breast were analyzed during shelf life. Results showed that the shelf life of chicken meat kept in films containing ZEO improved from three to nine days compared to the control sample without any direct contact with the film. Study of oxygen barrier properties of bilayer film without essential oils (0.096 cm³ μm/m² d kPa) in comparison with PP film (416 cm³ μm/m² d kPa) shows that coating of PP with CMC significantly reduces oxygen permeation of the obtained packaging (P<0.05), which reduced aerobic bacteria growth. Chemical composition of ZEO was also evaluated by gas chromatography–mass spectrometry (GC–MS), and this shows that thymol was the main antimicrobial and antioxidant component of the essential oil. The results revealed that PP/CMC containing ZEO has good potential for application as active food packaging in indirect contact which would also improve sensory properties of product.

Keywords: Shelf life, chicken breast, polypropylene, carboxymethyl cellulose, essential oil.

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Authors: Kazem Alirezalu, Javad Hesari, Zabihollah Nemati, Boukaga Farmani

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The application of natural plant extracts which are rich in promising antioxidants and antimicrobial ingredients in the production of frankfurter-type sausages addresses consumer demands for healthier, more functional meat products. The effects of olive leaves, green tea and Urtica dioica L. extracts on physicochemical, microbiological and sensory characteristic of frankfurter-type sausage were investigated during 45 days of storage at 4 °C. The results revealed that pH and phenolic compounds decreased significantly (P < 0.05) in all samples during storage. Sausages containing 500 ppm green tea extract (1.78 mg/kg) showed the lowest TBARS values compared to olive leaves (2.01 mg/kg), Urtica dioica L. (2.26 mg/kg) extracts and control (2.74 mg/kg). Plant extracts significantly (P < 0.05) reduced the count of total mesophilic bacteria, yeast and mold by at least 2 log cycles (CFU/g) than those of control samples. Sensory characteristics of texture showed no difference (P > 0.05) between sausage samples, but sausage containing Urtica dioica L. extract had the highest score regarding flavor, freshness odor, and overall acceptability. Based on the results, sausage containing plant extracts could have a significant impact on antimicrobial activity, antioxidant capacity, sensory score, and shelf life stability of frankfurter-type sausage.

Keywords: Antimicrobial, antioxidant, frankfurter-type sausage, green tea, olive oil, shelf life, Urtica dioica L.

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Authors: Jaruwan Chutrtong, Tanakwan Bussabun

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Study production of tempeh inoculums powder by freeze-drying comparison with dry at 50°C and the sun bask for developing efficient tempeh inoculums for tempeh producing. Rhizopus oligosporus in PDA slant cultures was incubated at 30^oC for 3-5 days until spores and mycelium. Preparation spores suspension with sterilized water and then count the number of started spores. Fill spores suspension in Rice flour and soy flour, mixed with water (in the ratio 10: 7), which is steamed and sterilized at 121°C 15min. Incubated at room temperature for 4 days, count number of spores. Then take the progressive infection and full spore dough to dry at 50°C, sun bask, and lyophilize. Grind to powder. Then pack in plastic bags, stored at 5°C. To investigate quality of inoculums which use different methods, tempeh was fermented every 4 weeks for 24 weeks of the experiment. The result found that rice flour is not suitable to use as raw material in the production of powdered spores.  Fungi can growth rarely. Less number of spores and requires more time than soy flour. For drying method, lyophilization is the least possible time. Samples from this method are very hard and very dark and harder to grind than other methods. Drying at 50°C takes longer time than lyophilization but can also set time use for drying. Character of the dry samples is hard solid and brown color, but can be grinded easier. The sun drying takes the longest time, can’t determine the exact time. When the spore powder was used to fermented tempeh immediately, product has similar characters as which use spores that was fresh prepared. The tempeh has normal quality. When spore powder stored at low temperature, tempeh from storage spore in weeks 4, 8 and 12 is still normal. Time spending in production was close to the production of fresh spores. After storage  spores for 16 and 20 weeks, tempeh is still normal but growth and sporulation were take longer time than usual (about 6 hours). At 24 week storage, fungal growth is not good, made tempeh looks inferior to normal color, also smell and texture.

Keywords: Freeze drying, preparation, spore powder, tempeh.

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Authors: Ayfer Kilicarslan, Kubra Onol, Sercan Basit, Muhlis Nezihi Saridede

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Chalcopyrite (CuFeS2) is the most common primary mineral used for the commercial production of copper. The low dissolution efficiency of chalcopyrite in sulfate media has prevented an efficient industrial leaching of this mineral in sulfate media. Ferric ions, bacteria, oxygen and other oxidants have been used as oxidizing agents in the leaching of chalcopyrite in sulfate and chloride media under atmospheric or pressure leaching conditions. Two leaching methods were studied to evaluate chalcopyrite (CuFeS2) dissolution in acid media. First, the conventional oxidative acid leaching method was carried out using sulfuric acid (H2SO4) and potassium dichromate (K2Cr2O7) as oxidant at atmospheric pressure. Second, microwave-assisted acid leaching was performed using the microwave accelerated reaction system (MARS) for same reaction media. Parameters affecting the copper extraction such as leaching time, leaching temperature, concentration of H2SO4 and concentration of K2Cr2O7 were investigated. The results of conventional acid leaching experiments were compared to the microwave leaching method. It was found that the copper extraction obtained under high temperature and high concentrations of oxidant with microwave leaching is higher than those obtained conventionally. 81% copper extraction was obtained by the conventional oxidative acid leaching method in 180 min, with the concentration of 0.3 mol/L K2Cr2O7 in 0.5M H2SO4 at 50 ºC, while 93.5% copper extraction was obtained in 60 min with microwave leaching method under same conditions.

Keywords: Extraction, copper, microwave-assisted leaching, chalcopyrite, potassium dichromate.

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Authors: Victor Emery David Jr, Jiang Wenchao, Yasinta John, Md. Sahadat Hossain

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Sludge originates from the process of treatment of wastewater. It is the byproduct of wastewater treatment containing concentrated heavy metals and poorly biodegradable trace organic compounds, as well as potentially pathogenic organisms (viruses, bacteria, etc.) which are usually difficult to treat or dispose of. China, like other countries, is no stranger to the challenges posed by increase of wastewater. Treatment and disposal of sludge has been a problem for most cities in China. However, this problem has been exacerbated by other issues such as lack of technology, funding, and other factors. Suitable methods for such climatic conditions are still unavailable for modern cities in China. Against this background, this paper seeks to describe the methods used for treatment and disposal of sludge from industries and suggest a suitable method for treatment and disposal in Chongqing/China. From the research conducted, it was discovered that the highest treatment rate of sludge in Chongqing was 10.08%. The industrial waste piping system is not separated from the domestic system. Considering the proliferation of industry and urbanization, there is a likelihood that the production of sludge in Chongqing will increase. If the sludge produced is not properly managed, this may lead to adverse health and environmental effects. Disposal costs and methods for Chongqing were also included in this paper’s analysis. Research showed that incineration is the most expensive method of sludge disposal in China/Chongqing. Subsequent research therefore considered optional alternatives such as composting. Composting represents a relatively cheap waste disposal method considering the vast population, current technology and economic conditions of Chongqing, as well as China at large.

Keywords: Sludge, disposal of sludge, treatment, industrial sludge, Chongqing, wastewater.

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Authors: Zonglin Yang, Tatsuya Akiyama, Kerry S. Williamson, Michael J. Franklin, Thiruvarangan Ramaraj

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Pseudomonas aeruginosa is an opportunistic pathogen that forms surface-associated microbial communities (biofilms) on artificial implant devices and on human tissue. Biofilm infections are difficult to treat with antibiotics, in part, because the bacteria in biofilms are physiologically heterogeneous. One measure of biological heterogeneity in a population of cells is to quantify the cellular concentrations of ribosomes, which can be probed with fluorescently labeled nucleic acids. The fluorescent signal intensity following fluorescence in situ hybridization (FISH) analysis correlates to the cellular level of ribosomes. The goals here are to provide computationally and statistically robust approaches to automatically quantify cellular heterogeneity in biofilms from a large library of epifluorescent microscopy FISH images. In this work, the initial steps were developed toward these goals by developing an automated biofilm detection approach for use with FISH images. The approach allows rapid identification of biofilm regions from FISH images that are counterstained with fluorescent dyes. This methodology provides advances over other computational methods, allowing subtraction of spurious signals and non-biological fluorescent substrata. This method will be a robust and user-friendly approach which will enable users to semi-automatically detect biofilm boundaries and extract intensity values from fluorescent images for quantitative analysis of biofilm heterogeneity.

Keywords: Image informatics, Pseudomonas aeruginosa, biofilm, FISH, computer vision, data visualization.

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Authors: Samad Lotfollahzadeh

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Escherichia coli (E. coli) is the most isolated bacteria from blood circulation of septicemic calves. Given the prevalence of septicemia in animals and its economic importance in veterinary practice, better understanding of changes in clinical signs following disease, may contribute to early detection of disorder. The present study has been carried out to detect changes of clinical signs in induced sepsis in calves with E. coli. Colisepticemia has been induced in 10 twenty-day old healthy Holstein- Frisian calves with intravenous injection of 1.5 X 109 colony forming units (cfu) of O111:H8 strain of E. coli. Clinical signs including rectal temperature, heart rate, respiratory rate, shock, appetite, sucking reflex, feces consistency, general behavior, dehydration and standing ability were recorded in experimental calves during 24 hours after induction of colisepticemia. Blood culture was also carried out from calves four times during experiment. ANOVA with repeated measure is used to see changes of calves’ clinical signs to experimental colisepticemia, and values of P≤ 0.05 was considered statistically significant. Mean values of rectal temperature and heart rate as well as median values of respiratory rate, appetite, suckling reflex, standing ability and feces consistency of experimental calves increased significantly during study (P<0.05). In the present study median value of shock score was not significantly increased in experimental calves (P> 0.05). The results of present study showed that total score of clinical signs in calves with experimental colisepticemia increased significantly, although score of some clinical signs such as shock did not change significantly.

Keywords: Calves, Clinical signs scoring, E. coli O111:H8, Experimental colisepticemia,

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Authors: J. Pazourek, K. Šmejkal, P. Kollár, J. Rajchard, J. Šinko, Z. Balounová, E. Vlková, H. Salmonová

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Pectinatella magnifica (Leidy, 1851) is an invasive freshwater animal that lives in colonies. A colony of Pectinatella magnifica (a gelatinous blob) can be up to several feet in diameter large and under favorable conditions it exhibits an extreme growth rate. Recently European countries around rivers of Elbe, Oder, Danube, Rhine and Vltava have confirmed invasion of Pectinatella magnifica, including freshwater reservoirs in South Bohemia (Czech Republic). Our project (Czech Science Foundation, GAČR P503/12/0337) is focused onto biology and chemistry of Pectinatella magnifica. We monitor the organism occurrence in selected South Bohemia ponds and sandpits during the last years, collecting information about physical properties of surrounding water, and sampling the colonies for various analyses (classification, maps of secondary metabolites, toxicity tests). Because the gelatinous matrix is during the colony lifetime also a host for algae, bacteria and cyanobacteria (co-habitants), in this contribution, we also applied a high performance liquid chromatography (HPLC) method for determination of potentially present cyanobacterial toxins (microcystin-LR, microcystin-RR, nodularin). Results from the last 3-year monitoring show that these toxins are under limit of detection (LOD), so that they do not represent a danger yet. The final goal of our study is to assess toxicity risks related to fresh water resources invaded by Pectinatella magnifica, and to understand the process of invasion, which can enable to control it.

Keywords: Cyanobacteria, freshwater resources, Pectinatella magnifica invasion, toxicity monitoring.

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Authors: Elhamalsadat Ghaffari, Moghan Amirhosseinian, Amir Khaleghipour

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Multifunctional bioactive glasses (BGs) are designed with a focus on the provision of bactericidal and biological properties desired for angiogenesis, osteogenesis, and ultimately potential applications in bone tissue engineering. To achieve these, six sol-gel copper/magnesium substituted derivatives of 58S-BG, i.e. a mol% series of 60SiO₂-4P₂O₅-5CuO-(31-x) CaO/xMgO (where x=0, 1, 3, 5, 8, and 10), were synthesized. Afterwards, the effect of MgO/CaO substitution on the in vitro formation of nano-hydroxyapatite (HA), osteoblast-like cell responses and BGs antibacterial performance were studied. During the BGs synthesis, the elimination of nitrates was achieved at 700 °C that prevented the BGs crystallization and stabilized the obtained dried gels. The structural and morphological evaluations were performed with X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). These characterizations revealed that Cu-substituted 58S-BG consisting of 5 mol% MgO (BG-5/5) slightly had retarded the formation of HA. In addition, Cu-substituted 58S-BGs consisting 8 mol% and 10 mol% MgO (BG-5/8 and BG-5/10) displayed lower bioactivity probably due to the lower ion release rate of Ca–Si into the simulated body fluid (SBF). The determination of 3-(4, 5 dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and alkaline phosphate (ALP) activities proved that the highest values of both differentiation and proliferation of MC3T3-E1 cells can be obtained from a 5 mol% MgO substituted BG, while the over addition of MgO (8 mol% and 10 mol%) decreased the bioactivity. Furthermore, these novel Cu/Mg-substituted 58S-BGs displayed antibacterial effect against methicillin-resistant Staphylococcus aureus bacteria. Taken together, the results suggest the equally-substituted BG-5/5 (i.e. the one consists of 5 mol% of both CuO and MgO) as a promising candidate for bone tissue engineering, among all newly designed BGs in this work, owing to its desirable cell proliferation, ALP activity and antibacterial properties.

Keywords: Apatite, bioactivity, biomedical applications sol-gel processes.

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Authors: A. Rajesh Kanna, G. Suresh Kumar, Sathyanaryana N. Gummadi

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There are various sources of energies available worldwide and among them, crude oil plays a vital role. Oil recovery is achieved using conventional primary and secondary recovery methods. In-order to recover the remaining residual oil, technologies like Enhanced Oil Recovery (EOR) are utilized which is also known as tertiary recovery. Among EOR, Microbial enhanced oil recovery (MEOR) is a technique which enables the improvement of oil recovery by injection of bio-surfactant produced by microorganisms. Bio-surfactant can retrieve unrecoverable oil from the cap rock which is held by high capillary force. Bio-surfactant is a surface active agent which can reduce the interfacial tension and reduce viscosity of oil and thereby oil can be recovered to the surface as the mobility of the oil is increased. Research in this area has shown promising results besides the method is echo-friendly and cost effective compared with other EOR techniques. In our research, on laboratory scale we produced bio-surfactant using the strain Pseudomonas putida (MTCC 2467) and injected into designed simple sand packed column which resembles actual petroleum reservoir. The experiment was conducted in order to determine the efficiency of produced bio-surfactant in oil recovery. The column was made of plastic material with 10 cm in length. The diameter was 2.5 cm. The column was packed with fine sand material. Sand was saturated with brine initially followed by oil saturation. Water flooding followed by bio-surfactant injection was done to determine the amount of oil recovered. Further, the injection of bio-surfactant volume was varied and checked how effectively oil recovery can be achieved. A comparative study was also done by injecting Triton X 100 which is one of the chemical surfactant. Since, bio-surfactant reduced surface and interfacial tension oil can be easily recovered from the porous sand packed column.

Keywords: Bio-surfactant, Bacteria, Interfacial tension, Sand column.

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Authors: Samy A. Selim

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The antimicrobial, antiplasmid and cytotoxic activities of marine algae Halimeda opuntia and Sarconema filiforme were investigated. Antimicrobial bioassay against some human pathogenic bacteria and yeast were conducted using disc diffusion method. Halimeda extract exhibited antibacterial activity against six species of microrganisms, with significant inhibition against Staphylococcus aureus. While Sarconema extract was better potent as antifungal against Candida albicans. Comparative antibacterial studies showed that Halimeda extract showed equivalent or better activity as compared with commercial antibiotic when tested against Staphylococcus aureus. Further tests conducted using dilution method showed both extracts as having bacteriostatic mode of action against the tested microorganisms. Methanol extract of two species showed significant cytotoxicity (LC50 <500μg) on brine shrimp. Halimeda opuntia showed highest cytotoxic activity (LC50 =192.3μg). Also, the present investigation was undertaken to investigate the ability of methanolic extract of the algal extracts to cure R-plasmids from certain clinical E. coli isolates. The active fraction of Halimeda and Sarconema could cure plasmids from E. coli at curing efficiencies of approximately 78%. The active fraction mediated plasmid curing resulted in the subsequent loss of antibiotic resistance encoded in the plasmids as revealed by antibiotic resistance profile of cured strains. The screening results confirm the possible use of marine algae Halimeda opuntia and Sarconema filiforme as a source of pharmacological benefits.

Keywords: Antimicrobial, antiplasmid Cytotoxicity, Marine Algae.

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Authors: Alireza Arab, Morteza Elsa, Amirhossein Moghanian

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In this study, experiments were carried out to achieve a promising multifunctional and modified silicate based bioactive glass (BG). The main aim of the study was investigating the effect of lithium (Li) and magnesium (Mg) substitution, on in vitro bioactivity of substituted-58S BG. Moreover, it is noteworthy to state that modified BGs were synthesized in 60SiO₂–(36-x)CaO–4P₂O₅–(x)Li₂O and 60SiO₂–(36-x)CaO–4P₂O₅–(x)MgO (where x = 0, 5, 10 mol.%) quaternary systems, by sol-gel method. Their performance was investigated through different aspects such as biocompatibility, antibacterial activity as well as their effect on alkaline phosphatase (ALP) activity, and proliferation of MC3T3 cells. The antibacterial efficiency was evaluated against methicillin-resistant Staphylococcus aureus bacteria. To do so, CaO was substituted with Li₂O and MgO up to 10 mol % in 58S-BGs and then samples were immersed in simulated body fluid up to 14 days and then, characterized by X-ray diffraction, Fourier transform infrared spectroscopy, inductively coupled plasma atomic emission spectrometry, and scanning electron microscopy. Results indicated that this modification led to a retarding effect on in vitro hydroxyapatite (HA) formation due to the lower supersaturation degree for nucleation of HA compared with 58s-BG. Meanwhile, magnesium revealed further pronounced effect. The 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) and ALP analysis illustrated that substitutions of both Li₂O and MgO, up to 5 mol %, had increasing effect on biocompatibility and stimulating proliferation of the pre-osteoblast MC3T3 cells in comparison to the control specimen. Regarding to bactericidal efficiency, the substitution of either Li or Mg for Ca in the 58s BG composition led to statistically significant difference in antibacterial behaviors of substituted-BGs. Meanwhile, the sample containing 5 mol % CaO/Li₂O substitution (BG-5L) was selected as a multifunctional biomaterial in bone repair/regeneration due to the improved biocompatibility, enhanced ALP activity and antibacterial efficiency among all of the synthesized L-BGs and M-BGs.

Keywords: Alkaline, alkaline earth, bioactivity, biomedical applications, sol-gel processes.

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Authors: J. L. Chukwuneke, C. H. Achebe, S. N. Omenyi

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This research work presents the surface thermodynamics approach to M-TB/HIV-Human sputum interactions. This involved the use of the Hamaker coefficient concept as a surface energetics tool in determining the interaction processes, with the surface interfacial energies explained using van der Waals concept of particle interactions. The Lifshitz derivation for van der Waals forces was applied as an alternative to the contact angle approach which has been widely used in other biological systems. The methodology involved taking sputum samples from twenty infected persons and from twenty uninfected persons for absorbance measurement using a digital Ultraviolet visible Spectrophotometer. The variables required for the computations with the Lifshitz formula were derived from the absorbance data. The Matlab software tools were used in the mathematical analysis of the data produced from the experiments (absorbance values). The Hamaker constants and the combined Hamaker coefficients were obtained using the values of the dielectric constant together with the Lifshitz Equation. The absolute combined Hamaker coefficients A132abs and A131abs on both infected and uninfected sputum samples gave the values of A132abs = 0.21631x10-21Joule for M-TB infected sputum and Ã132abs = 0.18825x10-21Joule for M-TB/HIV infected sputum. The significance of this result is the positive value of the absolute combined Hamaker coefficient which suggests the existence of net positive van der waals forces demonstrating an attraction between the bacteria and the macrophage. This however, implies that infection can occur. It was also shown that in the presence of HIV, the interaction energy is reduced by 13% conforming adverse effects observed in HIV patients suffering from tuberculosis.

Keywords: Absorbance, dielectric constant, Hamaker coefficient, Lifshitz formula, macrophage, Mycobacterium tuberculosis, Van der Waals forces.

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Authors: Alaa Hussein Wadi

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The principal objective of a water treatment plant is to produce water that satisfies a set of drinking water quality standards at a reasonable price to the consumers. The gravel-bed flocculator provide a simple and inexpensive design for flocculation in small water treatment plants (less than 5000 m3/day capacity). The packed bed of gravel provides ideal conditions for the formation of compact settleable flocs because of continuous recontact provided by the sinuous flow of water through the interstices formed by the gravel. The field data which were obtained from the operation of the water supply treatment unit cover the physical, chemical and biological water qualities of the raw and settled water as obtained by the operation of the treatment unit. The experiments were carried out with the aim of assessing the efficiency of the gravel filter in removing the turbidity, pathogenic bacteria, from the raw water. The water treatment plant, which was constructed for the treatment of river water, was in principle a rapid sand filter. The results show that the average value of the turbidity level of the settled water was 4.83 NTU with a standard deviation of turbidity 2.893 NTU. This indicated that the removal efficiency of the sedimentation tank (gravel filter) was about 67.8 %. for pH values fluctuated between 7.75 and 8.15, indicating the alkaline nature of the raw water of the river Shatt Al-Hilla, as expected. Raw water pH is depressed slightly following alum coagulation. The pH of the settled water ranged from 7.75 to a maximum of 8.05. The bacteriological tests which were carried out on the water samples were: total coliform test, E-coli test, and the plate count test. In each test the procedure used was as outlined in the Standard Methods for the Examination of Water and Wastewater (APHA, AWWA, and WPCF, 1985). The gravel filter exhibit a low performance in removing bacterial load. The percentage bacterial removal, which is maximum for total plate count (19%) and minimum for total coliform (16.82%).

Keywords: Gravel bed flocculator, turbidity, total coliform.

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Authors: P. Murphy, D. Vasic, A. W. Gunaratne, T. Tugonon, M. Ison, C. Pagonis, E. T. Sitchon, A. Le Busque, T. J. Borody

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Irritable bowel syndrome (IBS) is a gastrointestinal disorder characterized by an alteration in bowel movements. There are three different types of IBS: diarrhea-predominant IBS (IBS-D), constipation-predominant IBS (IBS-C) and IBS with mixed bowel habit (IBS-M). Antimicrobials are increasingly being used as treatment for all types of IBS. Due to this increased use and subsequent success, the gut microbiome as a factor in the etiology of IBS is becoming more apparent. Accepted standard treatment has focused on IBS-C and involves either vancomycin or rifaximin. Here, we report on a cohort of 18 patients treated with both vancomycin and rifaximin for IBS-D. These patients’ records were reviewed retrospectively. In this cohort, patients were aged between 24-74 years (mean 44 years) and nine were female. At baseline all patients had diarrhea, four with mucus and one with blood. Other reported symptoms include abdominal pain (n = 11) bloating (n = 9), flatulence (n = 7), fatigue (n = 4) and nausea (n = 3). Patient’s treatments were personalized according to their symptom severity and tolerability and were treated with a combination of rifaximin (500-3000 mg/d) and vancomycin (500 mg-1500 mg/d) for an ongoing period. Follow-ups were conducted between 2-32 weeks. Of all patients, 89% reported improvement of at least 1 symptom, one reported no change and one patient’s symptoms got worse. The success of this combination treatment could be due to the different mechanisms of action undertaken by each medication. Vancomycin works by inhibiting the cell wall of the bacteria and rifaximin by inhibiting protein synthesis. This success in treatment validates the idea that IBS-D may be driven by a bacterial infection of the gastrointestinal microbiome. As IBS-D presents similarly to Clostridium difficile and symptom improvement can occur with the same treatment as Clostridium difficile of rifaximin and vancomycin, there is reason to suggest that the infectious agent could be an unidentified strain of Clostridium. Although these results offer some validity to the theory, more research is required.

Keywords: Clostridium difficile infection, diarrhea predominant irritable bowel syndrome, microbiome, vancomycin/rifaximin combination.

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Authors: M. Naimi, M. B. Khaled

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The present study consisted of an applied test in meat system to assess the effectiveness of three bio agents bacteriocinproducing strains: Lm24: Lactobacillus sakei, Lm14and Lm25: Pediococcus spp. Two tests were carried out: The ex situ test was intended for three batches added with crude bacteriocin solutions at 12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the in situ one consisted of four batches; three of them inoculated with one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The two used tests were done in the presence of the pathogen St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch served as a positive or a negative control was used too. The incubation was performed at 7°C. Total viable counts, staphylococci and lactic acid bacteria, at the beginning and at selected times with interval of three days were enumerated. Physico-chemical determinations (except for in situ test): pH, dry mater, sugars, fat and total protein, at the beginning and at end of the experiment, were done, according to the international norms. Our results confirmed the ex situ effectiveness. Furthermore, the batches affected negatively the total microbial load over the incubation days, and showed a significant regression in staphylococcal load at day seven, for Lm14, Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed that, at the last day of culture, staphylococcal load was nil for the three batches. In the in situ test, the cultures displayed less inhibitory attitude and recorded a decrease in staphylococcal load, for Lm14, Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore, physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%, 0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%, 5.37%, 5.5%. This study suggests that the use of selected strains as Lm25 could lead to the best results and would help in preserving and extending the shelf life of lamb meat.

Keywords: Biocontrol, in situ and ex situ, meat system, St. aureus, Lactobacillus sakei, Pediococcus spp.

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Authors: Mohammad Abdollahi

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Tomato (Lycopersicon esculentum Mill.) is one of the most popular, widely grown and the second most important vegetable crop, after potatoes. Nematodes have been identified as one of the major pests affecting tomato production throughout the world. The most destructive nematodes are the genus Meloidogyne. Most widespread and devastating species of this genus are M. incognita, M. javanica, and M. arenaria. These species can cause complete crop loss under adverse growing conditions. There are several potential methods for management of the root knot nematodes. Although the chemicals are widely used against the phytonematodes, because of hazardous effects of these compounds on non-target organisms and on the environment, there is a need to develop other control strategies. Nowadays, non-chemical measures are widely used to control the plant parasitic nematodes. Biocontrol of phytonematodes is an important method among environment-friendly measures of nematode management. There are some soil-inhabiting fungi that have biocontrol potential on phytonematodes, which can be used in nematode management program. The fungus Metarhizium anisopliae, originally is an entomopathogenic bioagent. Biocontrol potential of this fungus on some phytonematodes has been reported earlier. Recently, use of organic soil amendments as well as the use of bioagents is under special attention in sustainable agriculture. This research aimed to reduce the pesticide use in control of root-knot nematode, Meloidogyne javanica in tomato. The effects of M. anisopliae IMI 330189 and different levels of oak tree debris on M. javanica were determined. The combination effect of the fungus as well as the different rates of soil amendments was determined. Pots were filled with steam pasteurized soil mixture and the six leaf tomato seedlings were inoculated with 3000 second stage larvae of M. javanica/kg of soil. After eight weeks, plant growth parameters and nematode reproduction factors were compared. Based on the results of our experiment, combination of M. anisopliae IMI 330189 and oak debris caused more than 90% reduction in reproduction factor of nematode, at the rates of 100 and 150 g/kg soil (P ≤ 0.05). As compared to control, the reduction in number of galls was 76%. It was 86% for nematode reproduction factor, showing the significance of combined effect of both tested agents. Our results showed that plant debris can increase the biological activity of the tested bioagent. It was also proved that there was no adverse effect of oak debris, which potentially has antimicrobial activity, on antagonistic power of applied bioagent.

Keywords: Biological control, nematode management, organic soil, Quercus branti, root knot nematode, soil amendment.

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Authors: Aka S. Koffi, N’Goran Yao, Philippe Bastide, Denis Bruneau, Diby Kadjo

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Cocoa beans (Theobroma cocoa L.) are the main components for chocolate manufacturing. The beans must be correctly fermented at first. Traditional process to perform the first fermentation (lactic fermentation) often consists in confining cacao beans using banana leaves or a fermentation basket, both of them leading to a poor product thermal insulation and to an inability to mix the product. Box fermenter reduces this loss by using a wood with large thickness (e>3cm), but mixing to homogenize the product is still hard to perform. Automatic fermenters are not rentable for most of producers. Heat (T>45°C) and acidity produced during the fermentation by microbiology activity of yeasts and bacteria are enabling the emergence of potential flavor and taste of future chocolate. In this study, a cylindro-rotative fermenter (FCR-V1) has been built and coconut fibers were used in its structure to confine heat. An axis of rotation (360°) has been integrated to facilitate the turning and homogenization of beans in the fermenter. This axis permits to put fermenter in a vertical position during the anaerobic alcoholic phase of fermentation, and horizontally during acetic phase to take advantage of the mid height filling. For circulation of air flow during turning in acetic phase, two woven rattan with grid have been made, one for the top and second for the bottom of the fermenter. In order to reduce air flow during acetic phase, two airtight covers are put on each grid cover. The efficiency of the turning by this kind of rotation, coupled with homogenization of the temperature, caused by the horizontal position in the acetic phase of the fermenter, contribute to having a good proportion of well-fermented beans (83.23%). In addition, beans’pH values ranged between 4.5 and 5.5. These values are ideal for enzymatic activity in the production of the aromatic compounds inside beans. The regularity of mass loss during all fermentation makes it possible to predict the drying surface corresponding to the amount being fermented.

Keywords: Cocoa fermentation, fermenter, microbial activity, temperature, turning.

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Authors: Maribel G. Guzmán, Jean Dille, Stephan Godet

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Silver nanoparticles were prepared by chemical reduction method. Silver nitrate was taken as the metal precursor and hydrazine hydrate as a reducing agent. The formation of the silver nanoparticles was monitored using UV-Vis absorption spectroscopy. The UV-Vis spectroscopy revealed the formation of silver nanopart├¡cles by exhibing the typical surface plasmon absorption maxima at 418-420 nm from the UV–Vis spectrum. Comparison of theoretical (Mie light scattering theory) and experimental results showed that diameter of silver nanoparticles in colloidal solution is about 60 nm. We have used energy-dispersive spectroscopy (EDX), X-ray diffraction (XRD), transmission electron microscopy (TEM) and, UV–Vis spectroscopy to characterize the nanoparticles obtained. The energy-dispersive spectroscopy (EDX) of the nanoparticles dispersion confirmed the presence of elemental silver signal no peaks of other impurity were detected. The average size and morphology of silver nanoparticles were determined by transmission electron microscopy (TEM). TEM photographs indicate that the nanopowders consist of well dispersed agglomerates of grains with a narrow size distribution (40 and 60 nm), whereas the radius of the individual particles are between 10 and 20 nm. The synthesized nanoparticles have been structurally characterized by X-ray diffraction and transmission high-energy electron diffraction (HEED). The peaks in the XRD pattern are in good agreement with the standard values of the face-centered-cubic form of metallic silver (ICCD-JCPDS card no. 4-0787) and no peaks of other impurity crystalline phases were detected. Additionally, the antibacterial activity of the nanopart├¡culas dispersion was measured by Kirby-Bauer method. The nanoparticles of silver showed high antimicrobial and bactericidal activity against gram positive bacteria such as Escherichia Coli, Pseudimonas aureginosa and staphylococcus aureus which is a highly methicillin resistant strain.

Keywords: Silver nanoparticles, surface plasmon, UV-Vis absorption spectrum, chemicals reduction.

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Authors: Deepak K. Semwal, Ruchi B. Semwal, Aijaz Ahmad, Guy P. Kamatou, Alvaro M. Viljoen

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Kigelia africana (Lam.) Benth. (Bignoniaceae) is a reputed traditional remedy for various human ailments such as skin diseases, microbial infections, melanoma, stomach troubles, metabolic disorders, malaria and general pains. In spite of the fruit being widely used for purposes related to its antibacterial and antifungal properties, the chemical constituents associated with the activity have not been fully identified. To elucidate the active principles, we evaluated the antimicrobial activity of fruit extracts and purified fractions against Staphylococcus aureus, Enterococcus faecalis, Moraxella catarrhalis, Escherichia coli, Candida albicans and Candida tropicalis. Shade-dried fruits were powdered and extracted with hydroalcoholic (1:1) mixture by soaking at room temperature for 72 h. The crude extract was further fractionated by column chromatography, with successive elution using hexane, dichloromethane, ethyl acetate, acetone and methanol. The dichloromethane and ethyl acetate fractions were combined and subjected to column chromatography to furnish a wax and oil from the eluates of 20% and 40% ethyl acetate in hexane, respectively. The GC-MS and GC×GC-MS results revealed that linoleic acid, linolenic acid, palmitic acid, arachidic acid and stearic acid were the major constituents in both oil and wax. The crude hydroalcoholic extract exhibited the strongest activity with MICs of 0.125-0.5 mg/mL, followed by the ethyl acetate (MICs = 0.125-1.0 mg/mL), dichloromethane (MICs = 0.250-2.0 mg/mL), hexane (MICs = 0.25- 2.0 mg/mL), acetone (MICs = 0.5-2.0 mg/mL) and methanol (MICs = 1.0-2.0 mg/mL), whereas the wax (MICs = 2.0-4.0 mg/mL) and oil (MICs = 4.0-8.0 mg/mL) showed poor activity. The study concludes that synergistic interactions of chemical constituents could be responsible for the antimicrobial activity of K. africana fruits, which needs a more holistic approach to understand the mechanism of its antimicrobial activity.

Keywords: Kigelia Africana, traditional medicine, antimicrobial activity, Candida albicans, palmitic acid, synergistic interaction.

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Authors: Suttijit Sriwatcharakul

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The bioactivity studies from the weed ethanolic crude extracts from leaf, stem, pod and root of wild spider flower; Cleoma viscosa Linn. were analyzed for the growth inhibition of 6 bacterial species; Salmonella typhimurium TISTR 5562, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus TISTR 1466, Streptococcus epidermidis ATCC 1228, Escherichia coli DMST 4212 and Bacillus subtilis ATCC 6633 with initial concentration crude extract of 50 mg/ml. The agar well diffusion results found that the extracts inhibit only gram positive bacteria species; S. aureus, S. epidermidis and B. subtilis. The minimum inhibition concentration study with gram positive strains revealed that leaf crude extract give the best result of the lowest concentration compared with other plant parts to inhibit the growth of S. aureus, S. epidermidis and B. subtilis at 0.78, 0.39 and lower than 0.39 mg/ml, respectively. The determination of total phenolic compounds in the crude extracts exhibited the highest phenolic content was 10.41 mg GAE/g dry weight in leaf crude extract. Analyzed the efficacy of free radical scavenging by using DPPH radical scavenging assay with all crude extracts showed value of IC50 of leaf, stem, pod and root crude extracts were 8.32, 12.26, 21.62 and 35.99 mg/ml, respectively. Studied cytotoxicity of crude extracts on human breast adenocarcinoma cell line by MTT assay found that pod extract had the most cytotoxicity CC50 value, 32.41 µg/ml. Antioxidant activity and cytotoxicity of crude extracts exhibited that the more increase of extract concentration, the more activities indicated. According to the bioactivities results, the leaf crude extract of Cleoma viscosa Linn. is the most interesting plant part for further work to search the beneficial of this weed.

Keywords: Antimicrobial, antioxidant activity, Cleoma viscosa Linn., cytotoxicity test, total phenolic compound.

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Authors: Avita K. Marihal, K.S. Jagadeesh, Sarita Sinha

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Pentachlorophenol (PCP) is a polychlorinated aromatic compound that is widespread in industrial effluents and is considered to be a serious pollutant. Among the variety of industrial effluents encountered, effluents from tanning industry are very important and have a serious pollution potential. PCP is also formed unintentionally in effluents of paper and pulp industries. It is highly persistent in soils and is lethal to a wide variety of beneficial microorganisms and insects, human beings and animals. The natural processes that breakdown toxic chemicals in the environment have become the focus of much attention to develop safe and environmentfriendly deactivation technologies. Microbes and plants are among the most important biological agents that remove and degrade waste materials to enable their recycling in the environment. The present investigation was carried out with the aim of developing a microbial system for bioremediation of PCP polluted soils. A number of plant species were evaluated for their ability to tolerate different concentrations of pentachlorophenol (PCP) in the soil. The experiment was conducted for 30 days under pot culture conditions. The toxic effect of PCP on plants was studied by monitoring seed germination, plant growth and biomass. As the concentration of PCP was increased to 50 ppm, the inhibition of seed germination, plant growth and biomass was also increased. Although PCP had a negative effect on all plant species tested, maize and groundnut showed the maximum tolerance to PCP. Other tolerating crops included wheat, safflower, sunflower, and soybean. From the rhizosphere soil of the tolerant seedlings, as many as twenty seven PCP tolerant bacteria were isolated. From soybean, 8; sunflower, 3; safflower 8; maize 2; groundnut and wheat, 3 each isolates were made. They were screened for their PCP degradation potentials. HPLC analyses of PCP degradation revealed that the isolate MAZ-2 degraded PCP completely. The isolate MAZ-1 was the next best isolate with 90 per cent PCP degradation. These strains hold promise to be used in the bioremediation of PCP polluted soils.

Keywords: Biodegradation, pentachlorophenol, rhizobacteria.

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Authors: Ana C. Sousa, Beatriz C. Santos, Fátima N. Serralha

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The Tagus estuary is heavily affected by industrial and urban activities, making bioremediation studies crucial for environmental preservation. Fuel contamination in the area can arise from various anthropogenic sources, such as oil spills from shipping, fuel storage and transfer operations, and industrial discharges. These pollutants can cause severe harm to the ecosystem and the organisms, including humans, that inhabit it. Nonetheless, there are always natural organisms with the ability to resist these pollutants and transform them into non-toxic or harmless substances, which defines the process of bioremediation. Exploring the microbial communities existing in soil and their capacity to break down hydrocarbons has the potential to enhance the development of more efficient bioremediation approaches. The aim of this investigation was to explore the existence of hydrocarbonoclastic microorganisms in six locations within the Tagus estuary, three on the north bank: Trancão River, Praia Fluvial do Cais das Colinas and Praia de Algés, and three on the south bank: Praia Fluvial de Alcochete, Praia Fluvial de Alburrica, and Praia da Trafaria. In all studied locations, native microorganisms of the genus Pseudomonas were identified. The bioremediation rate of common hydrocarbons like gasoline, hexane, and toluene was assessed using the redox indicator 2,6-dichlorophenolindophenol (DCPIP). Effective hydrocarbon-degrading bacterial strains were identified in all analyzed areas, despite adverse environmental conditions. The highest bioremediation rates were achieved for gasoline (68%) in Alburrica, hexane (65%) in Algés, and toluene (79%) in Algés. Generally, the bacteria demonstrated efficient degradation of hydrocarbons added to the culture medium, with higher rates of aerobic biodegradation of hydrocarbons observed. These findings underscore the necessity for further in situ studies to better comprehend the relationship between native microbial communities and the potential for pollutant degradation in soil.

Keywords: Biodegradability rate, hydrocarbonoclastic microorganisms, soil bioremediation, Tagus estuary.

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Authors: Shima Fasahat

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This research is an experimental research which was done about microbial fuel cells in order to study them for electricity generating and wastewater treatment. These days, it is very important to find new, clean and sustainable ways for energy supplying. Because of this reason there are many researchers around the world who are studying about new and sustainable energies. There are different ways to produce these kind of energies like: solar cells, wind turbines, geothermal energy, fuel cells and many other ways. Fuel cells have different types one of these types is microbial fuel cell. In this research, an MFC was built in order to study how it can be used for electricity generating and wastewater treatment. The microbial fuel cell which was used in this research is a reactor that has two tanks with a catalyst solution. The chemical reaction in microbial fuel cells is a redox reaction. The microbial fuel cell in this research is a two chamber MFC. Anode chamber is an anaerobic one (ABR reactor) and the other chamber is a cathode chamber. Anode chamber consists of stabilized sludge which is the source of microorganisms that do redox reaction. The main microorganisms here are: Propionibacterium and Clostridium. The electrodes of anode chamber are graphite pages. Cathode chamber consists of graphite page electrodes and catalysts like: O₂, KMnO₄ and C₆N₆FeK₄. The membrane which separates the chambers is Nafion117. The reason of choosing this membrane is explained in the complete paper. The main goal of this research is to generate electricity and treating wastewater. It was found that when you use electron receptor compounds like: O_2, MnO₄, C₆N₆FeK₄ the velocity of electron receiving speeds up and in a less time more current will be achieved. It was found that the best compounds for this purpose are compounds which have iron in their chemical formula. It is also important to pay attention to the amount of nutrients which enters to bacteria chamber. By adding extra nutrients in some cases the result will be reverse.  By using ABR the amount of chemical oxidation demand reduces per day till it arrives to a stable amount.

Keywords: Anaerobic baffled reactor, bioenergy, electrode, energy efficient, microbial fuel cell, renewable chemicals, sustainable.

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Authors: Lars A. L. Giske, Ola J. Mork, Emil Bjoerlykhaug

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The design of fish processing equipment greatly impacts how easy the cleaning process for the equipment is. This is a critical issue in fish processing, as cleaning of fish processing equipment is a task that is both costly and time consuming, in addition to being very important with regards to product quality. Even more, poorly cleaned equipment could in the worst case lead to contaminated product from which consumers could get ill. This paper will elucidate how equipment design changes could improve the work for the cleaners and saving money for the fish processing facilities by looking at a case for product design improvements. The design of fish processing equipment largely determines how easy it is to clean. “Design for cleaning” is the new hype in the industry and equipment where the ease of cleaning is prioritized gets a competitive advantage over equipment in which design for cleaning has not been prioritized. Design for cleaning is an important research area for equipment manufacturers. SeaSide AS is doing continuously improvements in the design of their products in order to gain a competitive advantage. The focus in this paper will be conveyors for internal logistic and a product called the “electro stunner” will be studied with regards to “Design for cleaning”. Often together with SeaSide’s customers, ideas for new products or product improvements are sketched out, 3D-modelled, discussed, revised, built and delivered. Feedback from the customers is taken into consideration, and the product design is revised once again. This loop was repeated multiple times, and led to new product designs. The new designs sometimes also cause the manufacturing processes to change (as in going from bolted to welded connections). Customers report back that the concrete changes applied to products by SeaSide has resulted in overall more easily cleaned equipment. These changes include, but are not limited to; welded connections (opposed to bolted connections), gaps between contact faces, opening up structures to allow cleaning “inside” equipment, and generally avoiding areas in which humidity and water may gather and build up. This is important, as there will always be bacteria in the water which will grow if the area never dries up. The work of creating more cleanable design is still ongoing, and will “never” be finished as new designs and new equipment will have their own challenges.

Keywords: Cleaning, design, equipment, fish processing, innovation.

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Authors: Oqba Basal, András Szabó

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Legume crops are able to fix atmospheric nitrogen by the symbiotic relation with specific bacteria, which allows the use of the mineral nitrogen-fertilizer to be reduced, or even excluded, resulting in more profit for the farmers and less pollution for the environment. Soybean (Glycine max (L.) Merrill) is one of the most important legumes with its high content of both protein and oil. However, it is recommended to combine the two nitrogen sources under stress conditions in order to overcome its negative effects. Drought stress is one of the most important abiotic stresses that increasingly limits soybean yields. A precise rate of mineral nitrogen under drought conditions is not confirmed, as it depends on many factors; soybean yield-potential and soil-nitrogen content to name a few. An experiment was conducted during 2017 growing season in Debrecen, Hungary to investigate the effects of nitrogen source on the physiology and the yield of the soybean cultivar 'Boglár'. Three N-fertilizer rates including no N-fertilizer (0 N), 35 kg ha^-1 of N-fertilizer (35 N) and 105 kg ha^-1 of N-fertilizer (105 N) were applied under three different irrigation regimes; severe drought stress (SD), moderate drought stress (MD) and control with no drought stress (ND). Half of the seeds in each treatment were pre-inoculated with Bradyrhizobium japonicum inoculant. The overall results showed significant differences associated with fertilization and irrigation, but not with inoculation. Increasing N rate was mostly accompanied with increased chlorophyll content and leaf area index, whereas it positively affected the plant height only when the drought was waived off. Plant height was the lowest under severe drought, regardless of inoculation and N-fertilizer application and rate. Inoculation increased the yield when there was no drought, and a low rate of N-fertilizer increased the yield furthermore; however, the high rate of N-fertilizer decreased the yield to a level even less than the inoculated control. On the other hand, the yield of non-inoculated plants increased as the N-fertilizer rate increased. Under drought conditions, adding N-fertilizer increased the yield of the non-inoculated plants compared to their inoculated counterparts; moreover, the high rate of N-fertilizer resulted in the best yield. Regardless of inoculation, the mean yield of the three fertilization rates was better when the water amount increased. It was concluded that applying N-fertilizer to provide the nitrogen needed by soybean plants, with the absence of N₂-fixation process, is very important. Moreover, adding relatively high rate of N-fertilizer is very important under severe drought stress to alleviate the drought negative effects. Further research to recommend the best N-fertilizer rate to inoculated soybean under drought stress conditions should be executed.

Keywords: Drought stress, inoculation, N-fertilizer, soybean physiology, yield.

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Authors: Tommaso Adamo, Gianpaolo Ghiani, Antonio D. Grieco, Emanuela Guerriero

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Batch production plants provide a wide range of scheduling problems. In pharmaceutical industries a batch process is usually described by a recipe, consisting of an ordering of tasks to produce the desired product. In this research work we focused on pharmaceutical production processes requiring the culture of a microorganism population (i.e. bacteria, yeasts or antibiotics). Several sources of uncertainty may influence the yield of the culture processes, including (i) low performance and quality of the cultured microorganism population or (ii) microbial contamination. For these reasons, robustness is a valuable property for the considered application context. In particular, a robust schedule will not collapse immediately when a cell of microorganisms has to be thrown away due to a microbial contamination. Indeed, a robust schedule should change locally in small proportions and the overall performance measure (i.e. makespan, lateness) should change a little if at all. In this research work we formulated a constraint programming optimization (COP) model for the robust planning of antibiotics production. We developed a discrete-time model with a multi-criteria objective, ordering the different criteria and performing a lexicographic optimization. A feasible solution of the proposed COP model is a schedule of a given set of tasks onto available resources. The schedule has to satisfy tasks precedence constraints, resource capacity constraints and time constraints. In particular time constraints model tasks duedates and resource availability time windows constraints. To improve the schedule robustness, we modeled the concept of (a, b) super-solutions, where (a, b) are input parameters of the COP model. An (a, b) super-solution is one in which if a variables (i.e. the completion times of a culture tasks) lose their values (i.e. cultures are contaminated), the solution can be repaired by assigning these variables values with a new values (i.e. the completion times of a backup culture tasks) and at most b other variables (i.e. delaying the completion of at most b other tasks). The efficiency and applicability of the proposed model is demonstrated by solving instances taken from a real-life pharmaceutical company. Computational results showed that the determined super-solutions are near-optimal.

Keywords: Constraint programming, super-solutions, robust scheduling, batch process, pharmaceutical industries.

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