Search results for: Escherichia coli bacteria

Authors: Zohreh Bayat, Dariush Minai-Tehrani

Abstract:

Lipases constitute one of the most important groups of industrial enzymes that catalyze the hydrolysis of triacylglycerol to glycerol and fatty acids. Muscarinic antagonist relieves smooth muscle spasm of the gastrointestinal tract and effect on the cardiovascular system. In this research, the effect of a muscarinic antagonist on the lipase activity of Pseudomonas aeruginosa was studied. Lineweaver–Burk plot showed that the drug inhibited the enzyme by competitive inhibition. The IC50 value (60 uM) and Ki (30 uM) of the drug revealed the drug bound to the enzyme with high affinity. Determination of enzyme activity in various pH and temperature showed that the maximum activity of lipase was at pH 8 and 60°C both in presence and absence of the drug.

Keywords: bacteria, inhibition, kinetics, lipase

Procedia PDF Downloads 432

Authors: Ayodele Odularu, Peter Ajibade, Albert Bolhuis

Abstract:

This study aimed at assessing the antibacterial activities of four zinc (II) complexes. Zinc (II) complexes of nitrogen, oxygen and sulfur coordination modes were synthesized using direct substitution reaction. The characterization techniques involved physicochemical properties (molar conductivity) and spectroscopic techniques. The molar conductivity gave the non-electrolytic nature of zinc (II) complexes. The spectral studies of zinc (II) complexes were done using electronic spectra (UV-Vis) and Fourier Transform Infra-red Spectroscopy (FT-IR). Spectral data from the spectroscopic studies confirmed the coordination of the mixed ligands with zinc (II) ion. The antibacterial activities of zinc(II) complexes of were all in supportive of Overtone’s concept and Tweedy’s theory of chelation for bacterial strains of S. aureus MRSA252 and E coli MC4100 because the zones of inhibition were greater than the corresponding ligands. In summary, all zinc (II) complexes of ZEPY, ZE1PH, ZE1PY and ZE135PY all have potentials for antibacterial activities.

Keywords: antibacterial activities, spectral studies, syntheses, zinc(II) complexes

Procedia PDF Downloads 252

Authors: Khosro Ghazvinian, Touba Khodaiean

Abstract:

Anaplasma organisms are obligatory intracellular bacteria belonging to the order Rickettsiales, family Anaplasmataceae. This disease is distributed around the globe and infected ticks are the most important vectors in anaplasmosis transmission. There is a little information about anaplasmosis in camels. This research investigated the blood films of 35 (20 male, 15 female) camels randomly selected from a flock of 150 camels. Samples were stained with Giemsa and Anaplasma sp. organisms were observed in six out of 35 (17.14 %) blood films. There were also some changes in Diff-Quick and morphology of leukocytes. No significant difference between male and female camels was observed (P>0.05). According to the results anaplasmosis is presented among camels in Iran.

Keywords: anaplasma, anaplasmosis, camel, Iran

Procedia PDF Downloads 232

Authors: Vanja Vlajkov, Ivana PajčIn, Mila Grahovac, Marta Loc, Dragana Budakov, Jovana Grahovac

Abstract:

The rhizosphere soil refers to the plant roots' dynamic environment characterized by their inhabitants' high biological activity. Rhizospheric bacteria are recognized as effective biocontrol agents and considered cardinal in alternative strategies for securing ecological plant diseases management. The need to suppress fungal pathogens is an urgent task, not only because of the direct economic losses caused by infection but also due to their ability to produce mycotoxins with harmful effects on human health. Aspergillus and Fusarium species are well-known producers of toxigenic metabolites with a high capacity to colonize crops and enter the food chain. The bacteria belonging to the Bacillus genus has been conceded as a plant beneficial species in agricultural practice and identified as plant growth-promoting rhizobacteria (PGPR). Besides incontestable potential, the full commercialization of microbial biopesticides is in the preliminary phase. Thus, there is a constant need for estimating the suitability of novel strains to be used as a central point of viable bioprocess leading to market-ready product development. In the present study, 76 potential producing strains were isolated from the rhizosphere soil, sampled from different localities in the Autonomous Province of Vojvodina, Republic of Serbia. The selective isolation process of strains started by resuspending 1 g of soil samples in 9 ml of saline and incubating at 28° C for 15 minutes at 150 rpm. After homogenization, thermal treatment at 100° C for 7 minutes was performed. Dilution series (10-1-10-3) were prepared, and 500 µl of each was inoculated on nutrient agar plates and incubated at 28° C for 48 h. The pure cultures of morphologically different strains indicating belonging to the Bacillus genus were obtained by the spread-plate technique. The cultivation of the isolated strains was carried out in an Erlenmeyer flask for 96 h, at 28 °C, 170 rpm. The antagonistic activity screening included two phytopathogenic fungi as test microorganisms: Aspergillus sp. and Fusarium sp. The mycelial growth inhibition was estimated based on the antimicrobial activity testing of cultivation broth by the diffusion method. For the Aspergillus sp., the highest antifungal activity was recorded for the isolates Kro-4a and Mah-1a. In contrast, for the Fusarium sp., following 15 isolates exhibited the highest antagonistic effect Par-1, Par-2, Par-3, Par-4, Kup-4, Paš-1b, Pap-3, Kro-2, Kro-3a, Kro-3b, Kra-1a, Kra-1b, Šar-1, Šar-2b and Šar-4. One-way ANOVA was performed to determine the antagonists' effect statistical significance on inhibition zone diameter. Duncan's multiple range test was conducted to define homogenous groups of antagonists with the same level of statistical significance regarding their effect on antimicrobial activity of the tested cultivation broth against tested pathogens. The study results have pointed out the significant in vitro potential of the isolated strains to be used as biocontrol agents for the suppression of the tested mycotoxigenic fungi. Further research should include the identification and detailed characterization of the most promising isolates and mode of action of the selected strains as biocontrol agents. The following research should also involve bioprocess optimization steps to fully reach the selected strains' potential as microbial biopesticides and design cost-effective biotechnological production.

Keywords: Bacillus, biocontrol, bioprocess, mycotoxigenic fungi

Procedia PDF Downloads 180

Authors: Fouzia Qamar, Shahida Hasnain

Abstract:

The objective of the present study was to evaluate the possible role of Agrobacterium tumefaciens as a possible insect biocontrol agent. Pests selected for the present challenge were adult males of Periplaneta americana and last instar larvae of Pieris brassicae and Spodoptera litura. Different ranges of bacterial doses were selected and tested to score the mortalities of the insects after 24 hours, for the lethal dose estimation studies. Mode of application for the inoculation of the bacteria, was the microinjection technique. The evaluation of the possible entomopathogenic carrying attribute of bacterial Ti plasmid, led to the conclusion that the loss of plasmid was associated with the loss of virulence against target insects.

Keywords: agrobacterium tumefaciens, toxicity assessment, biopesticidal attribute, entomopathogenic agent

Procedia PDF Downloads 358

Authors: Zuzana Sedrlova, Eva Slaninova, Ines Fritz, Christina Daffert, Stanislav Obruca

Abstract:

Cyanobacteria are ecologically extremely important phototrophic gram-negative bacteria capable of oxygenic photosynthesis. They synthesize many interesting metabolites such as glycogen, carotenoids, but the most interesting metabolites are polyhydroxyalkanoates (PHA). The main advantage of cyanobacteria is the fact they do not require costly organic substrate and, oppositely, cyanobacteria can fix CO₂. PHA serves primarily as a carbon and energy source and occurs in the form of intracellular granules in bacterial cells. It is possible, PHA helps cyanobacteria to survive stress conditions since increased PHA synthesis was observed during cultivation in stress conditions. PHA is microbial biopolymers that are biodegradable with similar properties as petrochemical synthetic plastics. Production of PHA by heterotrophic bacteria is expensive; for price reduction waste materials as input, materials are used. Positively, cyanobacteria principally do not require organic carbon substrate since they are capable of CO₂ fixation. In this work, we demonstrated that stress conditions lead to the highest obtained yields of PHA in cyanobacterial cultures. Two cyanobacterial cultures from genera Synechocystis were used in this work. Cultivations were performed either in Erlenmayer flask or in tube multicultivator. Multiple stressors were applied on cyanobacterial cultures, and stressors include PHA precursors. PHA precursors are chemical substances and some of them do not occur naturally in the environment. Cultivation with the same PHA precursors in the same concentration led to a 1,6x higher amount of PHA when a multicultivator was used. The highest amount of PHA reached 25 % of PHA in dry cyanobacterial biomass. Both strains are capable of co-polymer synthesis in the presence of their structural precursor. The composition of co-polymer differs in Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. Synechocystis sp. PCC 6803 cultivated with γ-butyrolakton accumulated co-polymer of 3-hydroxybutyrate (3HB) and 4-hydroxybutyrate (4HB) the composition of the copolymer was 56 % of 4HB and 44 % of 3HB. The total amount of PHA, as well as yield of biomass, was lower than in control due to the toxic properties of γ-butyrolakton. Funding: This study was partly funded by the project GA19- 19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), a project I 4082-B25. This work was supported by Brno, Ph.D. Talent – Funded by the Brno City Municipality.

Keywords: co-polymer, cyanobacteria, PHA, synechocystis

Procedia PDF Downloads 181

Authors: S. S. Mahlangu, R. K. K. Mbaya, D. D. Delport, H. Van. Zyl

Abstract:

The degrading effect due to bacterial growth on the structural integrity of concrete floor surfaces is predictable; this consequently cause development of surface micro cracks in which organisms penetrate through resulting in surface spalling. Hence, the need to develop mix design meeting the requirement of floor surfaces exposed to aggressive agent to improve certain material properties with good workability, extended lifespan and low cost is essential. In this work, tests were performed to examine the microbial activity on kitchen floor surfaces and the effect of adding admixtures. The biochemical test shows the existence of microorganisms (E.coli, Streptococcus) on newly casted structure. Of up to 6% porosity was reduced and improvement on structural integrity was observed upon adding mineral admixtures from the concrete mortar. The SEM result after 84 days of curing specimens, shows that chemical admixtures have significant role to enable retard bacterial penetration and good quality structure is achieved.

Keywords: admixture, organisms, porosity, strength

Procedia PDF Downloads 221

Authors: J. O. Alabi

Abstract:

Biogas is a clean burning, easily produced natural fuel that is an important source of energy for cooking and heating in rural areas and third world countries. Anaerobic bacteria inside biodigesters break down biomass to produce biogas. (Which is 70% methane)? Currently there is no simple way to compress and store biogas. So, in order to use biogas as a source of energy, a direct feed from biodigeser to the store tap or heater must be made. Any excess biogas is vented into the atmosphere, which is wasteful and car have a negative effect on the environment, we have been tasked with designing a system that will be able to compress biogas using an off-grid power supply, making the biogas portable and makes through the use of large-scale, shared biodigester. Our final design is a system that maximizes simplicity and safety while minimizing cost.

Keywords: biogas, biodigesters, natural fuel, bionanotechnology

Procedia PDF Downloads 343

Authors: Mokgadi Miranda Hlongwane, Ntebogeng Sharon Mokgalaka, Felix Dapare Dakora

Abstract:

Lessertia (L.) frutescens (syn. Sutherlandia frutescens) is a leguminous medicinal plant indigenous to South Africa. Traditionally, L. frutescens has been used to treat cancer, diabetes, epilepsy, fever, HIV, stomach problems, wounds and other ailments. This legume is endemic to the Cape fynbos, with large populations occurring wild and cultivated in the Cape Florist Region. Its widespread distribution in the Western Cape, Northern Cape, Eastern Cape and Kwazulu-Natal is linked to its increased use as a phytomedicine in the treatment of various diseases by traditional healers. The frequent harvesting of field plants for use as a medicine has made it necessary to undertake studies towards the conservation of Lessertia frutescens. As a legume, this species can form root nodules and fix atmospheric N₂ when in symbiosis with soil bacteria called rhizobia. So far, however, few studies (if any) have been done on the efficacy and diversity of native bacterial symbionts nodulating L. frutescens in South Africa. The aim of this project was to isolate and characterize L. frutescens-nodulating bacteria from five different locations in the Western Cape Province. This was done by trapping soil rhizobia using rhizosphere soil suspension to inoculate L. frutescens seedlings growing in sterilized sand and receiving sterile N-free Hoagland nutrient solution under glasshouse conditions. At 60 days after planting, root nodules were harvested from L. frutescens plants, surface-sterilized, macerated, and streaked on yeast mannitol agar (YMA) plates and incubated at 28 ˚C for observation of bacterial growth. The majority of isolates were slow-growers that took 6-14 days to appear on YMA plates. However, seven isolates were fast-growers, taking 2-4 days to appear on YMA plates. Single-colony cultures of the isolates were assessed for their ability to nodulate L. frutescens as a homologous host under glasshouse conditions. Of the 92 bacterial isolates tested, 63 elicited nodule formation on L. frutescens. Symbiotic effectiveness varied markedly between and among test isolates. There were also significant (p≤0.005) differences in nodulation, shoot biomass, photosynthetic rates, leaf transpiration and stomatal conductance of L. frutescens plants inoculated with the test isolates, which is an indication of their functional diversity.

Keywords: lessertia frutescens, nodulating, rhizobia, symbiotic effectiveness

Procedia PDF Downloads 171

Authors: F. A. Abdel-Mohdy, M. K. El-Bisi, A. Abou-Okeil, A. A. Sleem, S. El-Sabbagh, Kawther El-Shafei, Hoda S. El-Sayed, S. M. ElSawy

Abstract:

Chitosan/ polyacrylic acid membranes containing different amounts of Al2(SO4) and/or TiO2 were prepared. The prepared membranes were characterized by measuring mechanical properties, such as tensile strength and elongation at break, swelling properties, antimicrobial properties against gram-positive and gram-negative bacteria and blood clotting. The results obtained indicate that the presence of Al2(SO4) and TiO2 in the membrane formulations have an incremental effect on the antimicrobial properties and blood clotting in albino rate.

Keywords: Chitosan, acrylic acid, antibacterial, blood clotting, membrane

Procedia PDF Downloads 471

Authors: Karen Louise Lacey, Dario Javier Pavon Vargas, Massimiliano Rinaldi, Luca Cattani, Sara Rainieri

Abstract:

The effects of high-pressure processing (HPP) and thermal treatments (TT) on quality markers of strawberry nectar (12 °Brix, 3,3 pH) was studied before and after treatments. TT and HPP treatments ensured a 3-log aerobic bacteria inactivation. No significant difference was detected in terms of pH and °Brix. TT samples were less red (a* less positive) than all HPP treated samples, while all samples were less red than the control. Apparent viscosity was significantly increased in all the HPP treatments, at 10 1/s shear rate, control was 79.04±7.94 mPa•s and the 600 MPa-20 min treatment were 327.10±1.64 mPa•s. This work suggests that HPP treatments may maintain the quality markers of strawberry nectar better.

Keywords: HPP, strawberry nectar, colour , viscosity

Procedia PDF Downloads 106

Authors: Elmo Thiago Lins Cöuras Ford, Valentina Alessandra Carvalho do Vale

Abstract:

It presents a heating system using low cost alternative solar collectors to promote the disinfection of water in low income communities that take water contaminated by bacteria. The system consists of two solar collectors, with total area of 4 m² and was built using PET bottles and cans of beer and soft drinks. Each collector is made up of 8 PVC tubes, connected in series and work in continuous flow. It will determine the flux the most appropriate to generate the temperature to promote the disinfection. It will be presented results of the efficiency and thermal loss of system and results of analysis of water after undergoing the process of heating.

Keywords: Disinfection of water, solar heating system, poor communities, bioinformatics, biomedicine

Procedia PDF Downloads 465

Authors: Abdessemed Zineb, Atia Yahia, Yeza Salima

Abstract:

Water is self-purified by activated sludge process which makes its uniqueness. The main goal is the microbial biocenosis study of the input and output water of the waste water treatment system plant Timgad. 89.47% of the identified biocenosis belongs to ɤ-Proteobacteria while the remaining 10.52 % is equally divided between α-Proteobacteria and β-Proteobacteria. The antibiotics susceptibility profiles reveal that over 30 % are wild strains while the penicillinases are often present (11.30-20 %) with also other profiles. This proportion is worrying that the water discharged join the Oued Soltez used for irrigation. This disadvantage involves the installation of a chlorination step.

Keywords: activated sludge, biocenosis, antibiotics profiles, penicillinases, physic-chemical quality

Procedia PDF Downloads 283

Authors: Tariku Nefo Duke

Abstract:

Embelia is a genus of climbing shrubs in the family Myrsinaceae. Embelia schimperi is as important in traditional medicine as the other species in the genus. The plant has been much known as a local medicine for the treatment of tapeworms. In this project, extraction, phytochemical screening tests, isolation, and characterization of berries of the Embelia schimperi plant have been conducted. The chemical investigations of methanol and ethyl acetate (1:1) ratio extracts of the berries lead to the isolation of three new compounds. The compounds were identified to be alkaloids coded as AD, AN, and AG. Structural elucidations of the isolated compounds were accomplished using spectroscopic methods (IR, UV, ¹H NMR, ¹³C NMR, DEPT and 2D NMR, HPLC, and LC-MS). The alkaloid coded as (AN) has a wide MIC range of 6.31-25.46 mg/mL against all tested bacteria strains.

Keywords: Embelia schimper, HPLC, alkaloids, 2D NMR, MIC

Procedia PDF Downloads 79

Authors: Zexiao Zheng, Irene M. C. Lo

Abstract:

Organic pollutants, ammonia, and bacteria are major contaminants in sewage, which may adversely impact ecosystems without proper treatment. Conventional wastewater treatment plants (WWTPs) are operated to remove these contaminants from sewage but suffer from high carbon emissions and are powerless to remove emerging organic pollutants (EOPs). Herein, we have developed a low operational carbon emissions multifunctional photoelectrochemical (PEC) system for saline sewage treatment to simultaneously remove organic compounds, ammonia, and bacteria, coupled with H2 evolution. A reduced BiVO4 (r-BiVO4) with improved PEC properties due to the construction of oxygen vacancies and V4+ species was developed for the multifunctional PEC system. The PEC/r-BiVO4 process could treat saline sewage to meet local WWTPs’ discharge standard in 40 minutes at 2.0 V vs. Ag/AgCl and completely degrade carbamazepine (one of the EOPs), coupled with significant evolution of H2. A remarkable reduction in operational carbon emissions was achieved by the PEC/r-BiVO4 process compared with large-scale WWTPs, attributed to the restrained direct carbon emissions from the generation of greenhouse gases. Mechanistic investigation revealed that the PEC system could activate chloride ions in sewage to generate reactive chlorine species and facilitate •OH production, promoting contaminants removal. The PEC system exhibited operational feasibility at different pH and total suspended solids concentrations and has outstanding reusability and stability, confirming its promising practical potential. The study combined the simultaneous removal of three major contaminants from saline sewage and H2 evolution in a single PEC process, demonstrating a viable approach to supplementing and extending the existing wastewater treatment technologies. The study generated profound insights into the in-situ activation of existing chloride ions in sewage for contaminants removal and offered fundamental theories for applying the PEC system in sewage remediation with low operational carbon emissions. The developed PEC system can fit well with the future needs of wastewater treatment because of the following features: (i) low operational carbon emissions, benefiting the carbon neutrality process; (ii) higher quality of the effluent due to the elimination of EOPs; (iii) chemical-free in the operation of sewage treatment; (iv) easy reuse and recycling without secondary pollution.

Keywords: contaminants removal, H2 evolution, multifunctional PEC system, operational carbon emissions, saline sewage treatment, r-BiVO4 photoanodes

Procedia PDF Downloads 134

Authors: Mthokozisi B. C. Simelane

Abstract:

Despite the various efforts by governmental and non-governmental organizations aimed at eradicating the disease, malaria is said to kill a child every 30 seconds. Traditional healers use different concoctions prepared from medicinal plants to treat malaria. In the quest to bio-prospect plant-derived triterpenes for anti-malaria activity, we report here the in vivo antiplasmodial activity of ursolic acid acetate (ursolic acid isolated from dichloromethane extract of Mimusops caffra was chemically modified to its acetate derivative). The transdermal administration of ursolic acid acetate (UAA) dose dependently showed complete inhibition of the parasites’ growth at the highest concentration of 400 mg/kg after 15 days of Plasmodium berghei infection. UAA prevented the in vitro aggregation of MDH but did not prevent the expression of PfHsp 70 in E. coli XL1 blue cells. It, however, enhanced PfHsp70 ATPase activity with the specific activity of 65 units (amount of phosphate released 73.83 nmolPi/min.mg). Ursolic acid acetate prevented the formation of hemozoin (60 ± 0.02% at 6 mg/ml). The results suggest that Ursolic acid acetate possesses potential anti-malaria properties.

Keywords: Mimusops caffra, ursolic acid acetate, hemozoin, Malaria

Procedia PDF Downloads 407

Authors: Hattie Hope Makumbe, Bhekisisa Dlamini, Frederick Tabit

Abstract:

Listeria monocytogenes is one of the most important foodborne pathogens associated with ready-to-eat (RTE) food. This study investigated the incidence of Listeria monocytogenes in 80 RTE food sold in the formal (dairy and processed meat) and informal markets (vegetable salads, beef stew, and rice) of Johannesburg, South Africa. High Enterobacteriaceae, S. aureus, and E. coli counts were obtained, which ranged from 1.9-7.5 log CFU/g. Listeria monocytogenes microbial counts in the food samples ranged from 3.5-6.0 log colony forming unit per gram except in cooked rice. The Listeria monocytogenes isolates were identified using biochemical tests and confirmed with the Biolog identification system and PCR analyses. The percentage incidence for Listeria monocytogenes in ready to eat food was 12.5%. When Minimum Inhibitory Concentrations were under consideration, all disinfectants were effective against Listeria monocytogenes strains. For antimicrobial work, rates of resistance amongst the antibiotics ranged from 17-100%. Therefore, more effective preventive control strategies for Listeria monocytogenes are needed to reduce the prevalence of the pathogen in RTE food that is sold in Johannesburg.

Keywords: Listeria monocytogenes, Listeria species, ready to eat food, sanitiser efficacy

Procedia PDF Downloads 147

Authors: Borys Stegniy, Anton Gerilovych, Oleksii Solodiankin, Vitaliy Bolotin, Anton Stegniy, Denys Muzyka, Claudio Afonso

Abstract:

New APMV was isolated from white fronted goose in Ukraine. This isolate was tested serologically using monoclonal antibodies in haemagglutination-inhibition tests against APMV1-9. As the results obtained isolate showed cross reactions with APMV7. Following investigations were provided for the full genome sequencing using random primers and cloning into pCRII-TOPO. Analysis of 100 transformed colonies of E.coli using traditional sequencing gave us possibilities to find only 3 regions, which could identify by BLAST. The first region with the length of 367 bp had 70 % nucleotide sequence identity to the APMV 12 isolate Wigeon/Italy/3920_1/2005 at genome position 2419-2784. Next region (344 bp) had 66 % identity to the same APMV 12 isolate at position 4760-5103. The last region (365 bp) showed 71 % identity to Newcastle disease virus strain M4 at position 12569-12928.

Keywords: APMV, Newcastle disease virus, Ukraine, full genome sequencing

Procedia PDF Downloads 423

Authors: Elif Tugce Aksun Tumerkan, Chris Lowe, Hannah Krupa

Abstract:

Fluorescent proteins are self-sufficient in forming chromophores with a visible wavelength from 3 amino acids sequence within their own polypeptide structure. This chromophore – a molecule that absorbs a photon of light and exhibits an energy transition equal to the energy of the absorbed photon. Fluorescent proteins (FPs) consisted of a chain of 238 amino acid residues and composed of 11 beta strands shaped in a cylinder surrounding an alpha helix structure. A better understanding of the system of the chromospheres and the increasing advance in protein engineering in recent years, the properties of FPs offers the potential for new applications. They have used sensors and probes in molecular biology and cell-based research that giving a chance to observe these FPs tagged cell localization, structural variation and movement. For clarifying functional uses of fluorescent proteins, electrophoretic properties of these proteins are one of the most important parameters. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis is used for determining electrophoretic properties commonly. While there are many techniques are used for determining the functionality of protein-based research, SDS-PAGE analysis can only provide a molecular level assessment of the proteolytic fragments. Before SDS-PAGE analysis, fluorescent proteins need to successfully purified. Due to directly purification of the target, FPs is difficult from the animal, gene expression is commonly used which must be done by transformation with the plasmid. Furthermore, used gel within electrophoresis and staining agents properties have a key role. In this review, the different factors that have the impact on the electrophoretic properties of fluorescent proteins explored. Fluorescent protein separation and purification are the essential steps before electrophoresis that should be done very carefully. For protein purification, gene expression process and following steps have a significant function. For successful gene expression, the properties of selected bacteria for expression, used plasmid are essential. Each bacteria has own characteristics which are very sensitive to gene expression, also used procedure is the important factor for fluorescent protein expression. Another important factors are gel formula and used staining agents. Gel formula has an effect on the specific proteins mobilization and staining with correct agents is a key step for visualization of electrophoretic bands of protein. Visuality of proteins can be changed depending on staining reagents. Apparently, this review has emphasized that gene expression and purification have a stronger effect than electrophoresis protocol and staining agents.

Keywords: cell biology, gene expression, staining agents, SDS-page

Procedia PDF Downloads 171

Authors: Astrid Tannert, Anuradha Ramoji, Christina Ebert, Frederike Gladigau, Lorena Tuchscherr, Jürgen Popp, Ute Neugebauer

Abstract:

Staphylococcus aureus is a facultative intracellular pathogen, which by entering host cells may evade immunologic host response as well as antimicrobial treatment. In that way, S. aureus can cause persistent intracellular infections which are difficult to treat. Depending on the strain, S. aureus may persist at different intracellular locations like the phagolysosome. The first barrier invading pathogens from the blood stream that they have to cross are the endothelial cells lining the inner surface of blood and lymphatic vessels. Upon proceeding from an acute to a chronic infection, intracellular pathogens undergo certain biochemical and structural changes including a deceleration of metabolic processes to adopt for long-term intracellular survival and the development of a special phenotype designated as small colony variant. In this study, the endothelial cell line Ea.hy 926 was used as a model for acute and chronic S. aureus infection. To this end, Ea.hy 926 cells were cultured on QIAscout™ Microraft Arrays, a special graded cell culture substrate that contains around 12,000 microrafts of 200 µm edge length. After attachment to the substrate, the endothelial cells were infected with GFP-expressing S. aureus for 3 weeks. The acute infection and the development of persistent bacteria was followed by confocal laser scanning microscopy, scanning the whole Microraft Array for the presence and for detailed determination of the intracellular location of fluorescent intracellular bacteria every second day. After three weeks of infection representative microrafts containing infected cells, cells with protruded infections and cells that did never show any infection were isolated and fixed for Raman micro-spectroscopic investigation. For comparison, also microrafts with acute infection were isolated. The acquired Raman spectra are correlated with the fluorescence microscopic images to give hints about a) the molecular alterations in endothelial cells during acute and chronic infection compared to non-infected cells, and b) metabolic and structural changes within the pathogen when entering a mode of persistence within host cells. We thank Dr. Ruth Kläver from QIAGEN GmbH for her support regarding QIAscout technology. Financial support by the BMBF via the CSCC (FKZ 01EO1502) and from the DFG via the Jena Biophotonic and Imaging Laboratory (JBIL, FKZ PO 633/29-1, BA 1601/10-1) is highly acknowledged.

Keywords: correlative image analysis, intracellular infection, pathogen-host adaption, Raman micro-spectroscopy

Procedia PDF Downloads 165

Authors: Arjun Dakuri, J. Hayavadana

Abstract:

The study includes selection of 100 % bamboo fabric and cotton fabric for the study. The 100% bamboo fabrics were of 127 g/m², and 112 g/m² and 100% cotton grey fabric were of 104 g/m². The cotton fabric was desized, scoured, bleached and then treated with ZnO (as antimicrobial agent) with 1%, 2% and 3% using pad-dry cure method, whereas the bamboo fabrics were only desized. The antimicrobial activity of bamboo and ZnO treated cotton fabrics were evaluated and compared against E. coli and S. aureus as per the standard AATCC - 147. Moisture management properties of selected fabrics were also analyzed. Further, the selected fabric samples were tested for comfort properties like bending length, tearing strength, drape-ability, and specific handle force and air permeability. It was observed that bamboo fabrics show significant antibacterial activity and the same was shown by 3% ZnO treated cotton fabric. Both cotton and bamboo fabrics show improved moisture management properties than the cotton fabric. The comfort properties of bamboo fabrics are found to be superior to cotton fabrics making it more suitable for applications in place of cotton.

Keywords: antimicrobial activity, bamboo, cotton, comfort properties, moisture management, zinc oxide

Procedia PDF Downloads 329

Authors: Frans J. Smit, Wisdom A. Munzeiwa, Hermanus C. M. Vosloo, Lyn-Marie Birkholtz, Richard K. Haynes

Abstract:

Malaria and antimicrobial infections remain significant global health concerns, necessitating the continuous search for novel therapeutic approaches. This abstract presents an overview of the potential use of triazenes as effective agents against malaria and various antimicrobial pathogens. Triazenes are a class of compounds characterized by a linear arrangement of three nitrogen atoms, rendering them structurally distinct from their cyclic counterparts. This study investigates the efficacy of triazenes against malaria and explores their antimicrobial activity. Preliminary results revealed significant antimalarial activity of the triazenes, as evidenced by in vitro screening against P. falciparum, the causative agent of malaria. Furthermore, the compounds exhibited broad-spectrum antimicrobial activity, indicating their potential as effective antimicrobial agents. These compounds have shown inhibitory effects on various essential enzymes and processes involved in parasite survival, replication, and transmission. The mechanism of action of triazenes against malaria involves interactions with critical molecular targets, such as enzymes involved in the parasite's metabolic pathways and proteins responsible for host cell invasion. The antimicrobial activity of the triazenes against bacteria and fungi was investigated through disc diffusion screening. The antimicrobial efficacy of triazenes has been observed against both Gram-positive and Gram-negative bacteria, as well as multidrug-resistant strains, making them potential candidates for combating drug-resistant infections. Furthermore, triazenes possess favourable physicochemical properties, such as good stability, solubility, and low toxicity, which are essential for drug development. The structural versatility of triazenes allows for the modification of their chemical composition to enhance their potency, selectivity, and pharmacokinetic properties. These modifications can be tailored to target specific pathogens, increasing the potential for personalized treatment strategies. In conclusion, this study highlights the potential of triazenes as promising candidates for the development of novel antimalarial and antimicrobial therapeutics. Further investigations are necessary to determine the structure-activity relationships and optimize the pharmacological properties of these compounds. The results warrant additional research, including MIC studies, to further explore the antimicrobial activity of the triazenes. Ultimately, these findings contribute to the development of more effective strategies for combating malaria and microbial infections.

Keywords: malaria, anti-microbials, triazene, resistance

Procedia PDF Downloads 82

Authors: Andrea M. Rivas-Castillo, Marlenne Gómez-Ramirez, Isela Rodríguez-Pozos, Norma G. Rojas-Avelizapa

Abstract:

Spent catalysts are considered as hazardous residues of major concern, mainly due to the simultaneous presence of several metals in elevated concentrations. Although hydrometallurgical, pyrometallurgical and chelating agent methods are available to remove and recover some metals contained in spent catalysts; these procedures generate potentially hazardous wastes and the emission of harmful gases. Thus, biotechnological treatments are currently gaining importance to avoid the negative impacts of chemical technologies. To this end, diverse microorganisms have been used to assess the removal of metals from spent catalysts, comprising bacteria, archaea and fungi, whose resistance and metal uptake capabilities differ depending on the microorganism tested. Acidophilic sulfur oxidizing bacteria have been used to investigate the biotreatment and extraction of valuable metals from spent catalysts, namely Acidithiobacillus thiooxidans and Acidithiobacillus ferroxidans, as they present the ability to produce leaching agents such as sulfuric acid and sulfur oxidation intermediates. In the present work, the ability of A. thiooxidans DSM 26636 for the bioleaching of metals contained in five different spent catalysts was assessed by growing the culture in modified Starkey mineral medium (with elemental sulfur at 1%, w/v), and 1% (w/v) pulp density of each residue for up to 21 days at 30 °C and 150 rpm. Sulfur-oxidizing activity was periodically evaluated by determining sulfate concentration in the supernatants according to the NMX-k-436-1977 method. The production of sulfuric acid was assessed in the supernatants as well, by a titration procedure using NaOH 0.5 M with bromothymol blue as acid-base indicator, and by measuring pH using a digital potentiometer. On the other hand, Inductively Coupled Plasma - Optical Emission Spectrometry was used to analyze metal removal from the five different spent catalysts by A. thiooxidans DSM 26636. Results obtained show that, as could be expected, sulfuric acid production is directly related to the diminish of pH, and also to highest metal removal efficiencies. It was observed that Al and Fe are recurrently removed from refinery spent catalysts regardless of their origin and previous usage, although these removals may vary from 9.5 ± 2.2 to 439 ± 3.9 mg/kg for Al, and from 7.13 ± 0.31 to 368.4 ± 47.8 mg/kg for Fe, depending on the spent catalyst proven. Besides, bioleaching of metals like Mg, Ni, and Si was also obtained from automotive spent catalysts, which removals were of up to 66 ± 2.2, 6.2±0.07, and 100±2.4, respectively. Hence, the data presented here exhibit the potential of A. thiooxidans DSM 26636 for the simultaneous bioleaching of metals contained in spent catalysts from diverse provenance.

Keywords: bioleaching, metal removal, spent catalysts, Acidithiobacillus thiooxidans

Procedia PDF Downloads 123

Authors: Magdalena Diaka, Paweł Mazierski, Joanna Żebrowska, Michał Winiarski, Tomasz Klimczuk, Adriana Zaleska-Medynska

Abstract:

During the last years, TiO2 nanotubes have been widely studied due to their unique highly ordered array structure, unidirectional charge transfer and higher specific surface area compared to conventional TiO2 powder. These photoactive materials, in the form of thin layer, can be activated by low powered and low cost irradiation sources (such as LEDs) to remove VOCs, microorganism and to deodorize air streams. This is possible due to their directly growth on a support material and high surface area, which guarantee enhanced photon absorption together with an extensive adsorption of reactant molecules on the photocatalyst surface. TiO2 nanotubes exhibit also lots of other attractive properties, such as potential enhancement of electron percolation pathways, light conversion, and ion diffusion at the semiconductor-electrolyte interface. Pure TiO2 nanotubes were previously used to remove organic compounds from the gas phase as well as in water splitting reaction. The major factors limiting the use of TiO2 nanotubes, which have not been fully overcome, are their relatively large band gap (3-3,2 eV) and high recombination rate of photogenerated electron–hole pairs. Many different strategies were proposed to solve this problem, however titania nanostructures containing incorporated metal oxides like Ag2O shows very promising, new optical and photocatalytic properties. Unfortunately, there is still very limited number of reports regarding application of TiO2/MxOy nanostructures. In the present work, we prepared TiO2/Ag2O nanotubes obtained by anodization of Ti-Ag alloys containing 5, 10 and 15 wt. % Ag. Photocatalysts prepared in this way were characterized by X-ray diffraction spectroscopy (XRD), scanning electron microscopy (SEM), luminescence spectroscopy and UV-Vis spectroscopy. The activities of new TiO2/Ag2O were examined by photocatalytic degradation of toluene in gas phase reaction and phenol in aqueous phase using 1000 W Xenon lamp (Oriel) and light emitting diodes (LED) as a irradiation sources. Additionally efficiency of bacteria (Pseudomonas aeruginosa) removal from the gas phase was estimated. The number of surviving bacteria was determined by the serial twofold dilution microtiter plate method, in Tryptic Soy Broth medium (TSB, GibcoBRL).

Keywords: photocatalysis, antibacterial properties, titania nanotubes, new TiO2/MxOy nanostructures

Procedia PDF Downloads 279

Authors: Shikha Rani, Neeta Sehgal, Vipin Kumar Verma, Om Prakash

Abstract:

Introduction: Fish is an important protein source for humans and has great economic value. Fish cultures are affected due to various anthropogenic activities that lead to bacterial and viral infections. Aeromonas hydrophila is a fish pathogenic bacterium that causes several aquaculture outbreaks throughout the world and leads to huge mortalities. In this study, plants of no commercial value were used to investigate their immunostimulatory, antioxidant, anti-inflammatory, anti-bacterial, and disease resistance potential in fish against Aeromonas hydrophila, through fish feed fortification. Methods: The plant was dried at room temperature in the shade, dissolved in methanol, and analysed for biological compounds through GC-MS/MS. DPPH, FRAP, Phenolic, and flavonoids were estimated following standardized protocols. In silico molecular docking was also performed to validate its broad-spectrum activities based on binding affinity with specific proteins. Fish were divided into four groups (n=6; total 30 in a group): Group 1, non-challenged fish (fed on a non-supplemented diet); Group 2, fish challenged with bacteria (fed on a non-supplemented diet); Group 3 and 4, fish challenged with bacteria (A. hydrophila) and fed on plant supplemented feed at 2.5% and 5%. Blood was collected from the fish on 0, 7th, 14th, 21st, and 28th days. Serum was separated for glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase assay (ALP), lysozyme activity assay, superoxide dismutase assay (SOD), lipid peroxidation assay (LPO) and molecular parameters (including cytokine levels) were estimated through ELISA. The phagocytic activity of macrophages from the spleen and head kidney, along with quantitative analysis of immune-related genes, were analysed in different tissue samples. The digestive enzymes (Pepsin, Trypsin, and Chymotrypsin) were also measured to evaluate the effect of plant-supplemented feed on freshwater fish. Results and Discussion: GC-MS/MS analysis of a methanolic extract of plant validated the presence of key compounds having antioxidant, anti-inflammatory, anti-bacterial, anti-inflammatory, and immunomodulatory activities along with disease resistance properties. From biochemical investigations like ABTS, DPPH, and FRAP, the amount of total flavonoids, phenols, and promising binding affinities towards different proteins in molecular docking analysis helped us to realize the potential of this plant that can be used for investigation in the supplemented feed of fish. Measurement liver function tests, ALPs, oxidation-antioxidant enzyme concentrations, and immunoglobulin concentrations in the experimental groups (3 and 4) showed significant improvement as compared to the positive control group. The histopathological evaluation of the liver, spleen, and head kidney supports the biochemical findings. The isolated macrophages from the group fed on supplemented feed showed a higher percentage of phagocytosis and a phagocytic index, indicating an enhanced cell-mediated immune response. Significant improvements in digestive enzymes were also observed in fish fed on supplemented feed, even after weekly challenges with bacteria. Hence, the plant-fortified feed can be recommended as a regular feed to enhance fish immunity and disease resistance against the Aeromonas hydrophila infection after confirmation from the field trial.

Keywords: immunostimulation, antipathogen, plant fortified feed, macrophages, GC-MS/MS, in silico molecular docking

Procedia PDF Downloads 63

Authors: A. Dadda, D. Khelef, K. Ait-Oudia, R. Kaidi

Abstract:

Neonatal calf diarrhea is the most important disease of neonatal calves and results in the greatest economic losses due to disease in this age group in both dairy and beef calves. The objectives of the present study were to estimate the morbidity and the mortality of neonatal diarrhea in dairy calves also to determine aetiology and risk factors were caused diarrhea in dairy veal under 60 days old. A total of 324 claves, housed in 30 dairy breeding were followed during two velage season from January to Juan 2013. The total mortality was 5,9% and was significantly higher in calves had less than 15 days of age. The incidence rate of diarrhea was 31,5% and peaked in the first two weeks after velage. The main causes were breeding controls, defect of passive immunity, old of calf, production season, and nutrient of pregnant cattle, veal’s housing and infectious agents. ELISA test on 22 fecal samples revealed that the 31, 82% of dairy breeding were infected, by cryptosporidium parvum in 13, 6% of study population, E.Coli F5 in 9% and Rotavirus with rate of 4, 5%.

Keywords: diarrhoea, neonatal, mortality, aetiology, risk factors, incidence

Procedia PDF Downloads 618

Authors: Abdulrazzaq Hammal

Abstract:

In this study, Syrian sand was used to create ceramic membranes. The process of preparing the membranes involved several steps, starting with the purification of the studied sand using hydrochloric acid, sorting according to granular size, and mixing the sand with liquid sodium silicates as a binder. Next, the effects of binder ratio, pressure formation, treatment temperature, and sand grain size were studied. Further, nanoparticles of silver were added to the formed membranes to improve their ability to purify bacterially polluted water. Prepared membranes were quite successful in removing bacteria and chemicals from water, and the water's requirements were brought up to level with Syrian drinking water standards.

Keywords: ceramic, membrane, water, wastewater

Procedia PDF Downloads 47

Authors: Gousilin Leandra Rocha Da Silva, Stéfani T. A. Dantas, Bruna F. Rossi, Erika R. Bonsaglia, Ivana G. Castilho, Terue Sadatsune, Ary Fernandes Júnior, Vera l. M. Rall

Abstract:

Kidney failure causes decreased diuresis and accumulation of nitrogenous substances in the body. To increase patient survival, hemodialysis is used as a partial substitute for renal function. However, contamination of the water used in this treatment, causing bacteremia in patients, is a worldwide concern. The Burkholderia cepacia complex (Bcc), a group of bacteria with more than 20 species, is frequently isolated from hemodialysis water samples and comprises opportunistic bacteria, affecting immunosuppressed patients, due to its wide variety of virulence factors, in addition to innate resistance to several antimicrobial agents, contributing to the permanence in the hospital environment and to the pathogenesis in the host. The objective of the present work was to characterize molecularly and phenotypically Bcc isolates collected from the water and dialysate of the Hemodialysis Unit and from the blood of patients at a Public Hospital in Botucatu, São Paulo, Brazil, between 2019 and 2021. We used 33 Bcc isolates, previously obtained from blood cultures from patients with bacteremia undergoing hemodialysis treatment (2019-2021) and 24 isolates obtained from water and dialysate samples in a Hemodialysis Unit (same period). The recA gene was sequenced to identify the specific species among the Bcc group. All isolates were tested for the presence of some genes that encode virulence factors such as cblA, esmR, zmpA and zmpB. Considering the epidemiology of the outbreak, the Bcc isolates were molecularly characterized by Multi Locus Sequence Type (MLST) and by pulsed-field gel electrophoresis (PFGE). The verification and quantification of biofilm in a polystyrene microplate were performed by submitting the isolates to different incubation temperatures (20°C, average water temperature and 35°C, optimal temperature for group growth). The antibiogram was performed with disc diffusion tests on agar, using discs impregnated with cefepime (30µg), ceftazidime (30µg), ciprofloxacin (5µg), gentamicin (10µg), imipenem (10µg), amikacin 30µg), sulfametazol/trimethoprim (23.75/1.25µg) and ampicillin/sulbactam (10/10µg). The presence of ZmpB was identified in all isolates, while ZmpA was observed in 96.5% of the isolates, while none of them presented the cblA and esmR genes. The antibiogram of the 33 human isolates indicated that all were resistant to gentamicin, colistin, ampicillin/sulbactam and imipenem. 16 (48.5%) isolates were resistant to amikacin and lower rates of resistance were observed for meropenem, ceftazidime, cefepime, ciprofloxacin and piperacycline/tazobactam (6.1%). All isolates were sensitive to sulfametazol/trimethoprim, levofloxacin and tigecycline. As for the water isolates, resistance was observed only to gentamicin (34.8%) and imipenem (17.4%). According to PFGE results, all isolates obtained from humans and water belonged to the same pulsotype (1), which was identified by recA sequencing as B. cepacia¸, belonging to sequence type ST-767. By observing a single pulse type over three years, one can observe the persistence of this isolate in the pipeline, contaminating patients undergoing hemodialysis, despite the routine disinfection of water with peracetic acid. This persistence is probably due to the production of biofilm, which protects bacteria from disinfectants and, making this scenario more critical, several isolates proved to be multidrug-resistant (resistance to at least three groups of antimicrobials), turning the patient care even more difficult.

Keywords: hemodialysis, burkholderia cepacia, PFGE, MLST, multi drug resistance

Procedia PDF Downloads 78

Authors: Nidhi Nalin

Abstract:

This study is theoretical modelling of the fermentation process of glucose in agricultural wastes like discarded peaches to produce hydrogen, acetic acid, and carbon dioxide using Thermotoga neopolitana bacteria. The hydrogen gas produced in this process can be used in hydrogen fuel cells to generate power, and the fermented broth with acetic acid and salts could be utilized as salty vinegar if enough acetic acid is produced. The theoretical modelling was done using SuperPro software, and the results indicated how much sugar (discarded peaches) is required to produce both hydrogen and vinegar for the process to be profitable.

Keywords: fermentation, thermotoga, hydrogen, vinegar, biofuel

Procedia PDF Downloads 136

Authors: Agostinho Antunes

Abstract:

The completion of the human genome sequencing in 2003 opened a new perspective into the importance of whole genome sequencing projects, and currently multiple species are having their genomes completed sequenced, from simple organisms, such as bacteria, to more complex taxa, such as mammals. This voluminous sequencing data generated across multiple organisms provides also the framework to better understand the genetic makeup of such species and related ones, allowing to explore the genetic changes underlining the evolution of diverse phenotypic traits. Here, recent results from our group retrieved from comparative evolutionary genomic analyses of varied species will be considered to exemplify how gene novelty and gene enhancement by positive selection might have been determinant in the success of adaptive radiations into diverse habitats and lifestyles.

Keywords: adaptation, animals, evolution, genomics

Procedia PDF Downloads 411