Search results for: bacterial inoculum

Authors: Mohd Adil, Rosina Khan, Asad U. Khan, Vasantha Rupasinghe HP

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The study examined the effects of Syzygium aromaticum extracts on the virulence properties of Streptococcus mutans. The activity of glucosyltransferases in the presence of crude and diethylether fraction was reduced to 80% at concentration 78.12μg/ml and 39.06μg/ml respectively. The glycolytic pH drop by S. mutans cells was also disrupted by these extracts without affecting the bacterial viability. Microscopic analysis revealed morphological changes of the S. mutans biofilms, indicating that these plant extracts at sub-MICs could significantly affect the ability of S. mutans to form biofilm with distorted extracellular matrix. Furthermore, with the help of quantitative RT-PCR, the expression of different genes involved in adherence, quorum sensing, in the presence of these extracts were down regulated. The crude and active fractions were found effective in preventing caries development in rats. The data showed that S. aromaticum holds promise as a naturally occurring source of compounds that may prevent biofilm-related oral diseases.

Keywords: biofilm, quorum sensing, Streptococcus mutans, Syzygium aromaticum extract

Procedia PDF Downloads 288

Authors: M. Mehani, N. Salhi, T. Valeria, S. Ladjel

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Red River Gum (Eucalyptus camaldulensis) is a tree of the genus Eucalyptus widely distributed in Algeria and in the world. The value of its aromatic secondary metabolites offers new perspectives in the pharmaceutical industry. This strategy can contribute to the sustainable development of our country. Preliminary tests performed on the essential oil of Eucalyptus camendulensis showed that this oil has antibacterial activity vis-à-vis the bacterial strains (Enterococcus feacalis, Enterobacter cloaceai, Proteus microsilis, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa) and antifungic (Fusarium sporotrichioide and Fusarium graminearum). The culture medium used was nutrient broth Muller Hinton. The interaction between the bacteria and the essential oil is expressed by a zone of inhibition with diameters of MIC indirectly expression of. And we used the PDA medium to determine the fungal activity. The extraction of the aromatic fraction (essentially oil- hydrolat) of the fresh aerian part of the Eucalyptus camendulensis was performed by hydrodistillation. The average essential oil yield is 0.99%. The antimicrobial and fungal study of the essential oil and hydrosol showed a high inhibitory effect on the growth of pathogens.

Keywords: essential oil, Eucalyptus camendulensis, bacteria and fungi, red river gum

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Authors: Ravi J. Shah

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The present study deals with the synthesis of novel spiro[azetidine-2, 3’-indole]-2’, 4(1’H)-dione derivative from the reactions of 3-(phenylimino)-1,3-dihydro-2H-indol-2-one derivatives with chloracetyl chloride in presence of triethyl amine (TEA). All the compounds were characterized using IR, 1H NMR, MS and elemental analysis. They were screened for their antibacterial and antifungal activities. Results revealed that, compounds (7a), (7b), (7c), (7d) and (7e) showed very good activity with MIC value of 6.25-12.5 μg/ml against three evaluated bacterial strains and the remaining compounds showed good to moderate activity comparable to standard drugs as antibacterial agents. Compounds (7c) and (7h) displayed equipotent antifungal activity in comparison to standard drugs. Structure-activity relationship study of the compounds showed that the presence of electron withdrawing group substitution at 5’ and 7’ positions of indoline ring and on ortho or para position of phenyl ring increases both antibacterial and antifungal activity of the compound. Henceforth, our findings will have a good impact on chemists and biochemists for further investigations in search of bromine containing spiro fused antimicrobial agents.

Keywords: antibacterial activity, antifungal activity, 2-Azetidinone, indoline

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Authors: Fathiah Mohamed Zuki, Robert George Edyvean

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Electrostatic interaction energy (∆EEDL) is a part of the Extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory, which, together with van der Waals (∆EVDW) and acid base (∆EAB) interaction energies, has been extensively used to investigate the initial adhesion of bacteria to surfaces. Electrostatic or electrical double layer interaction energy is considerably affected by surface potential, however it cannot be determined experimentally and is usually replaced by zeta (ζ) potential via electrophoretic mobility. This paper focuses on the effect of ionic concentration as a function of pH and the effect of mineral grain size on ζ potential. It was found that both ionic strength and mineral grain size play a major role in determining the value of ζ potential for the adhesion of P. putida to hematite and quartz surfaces. Higher ζ potential values lead to higher electrostatic interaction energies and eventually to higher total XDLVO interaction energy resulting in bacterial repulsion.

Keywords: XDLVO, electrostatic interaction energy, zeta potential, P. putida, mineral

Procedia PDF Downloads 429

Authors: M. Peña Aguilar Juan, E. Nava Galván Claudia, Pastrana Palma Alberto

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In this work was developed and implemented a thermal fogging disinfection system to counteract pathogens from poultry feces in agribusiness farms, to reduce mortality rates and increase biosafety in them. The control system consists of two phases for the conditioning of the farm during the sanitary break. In the first phase, viral and bacterial inactivation was performed by treating the stool dry cleaning, along with the development of a specialized product that foster the generation of temperatures above 55 °C in less than 24 hr, for virus inactivation. In the second phase, a process for disinfection by fogging was implemented, along with the development of a specialized disinfectant that guarantee no risk for the operators’ health or birds. As a result of this process, it was possible to minimize the level of mortality of chickens on farms from 12% to 5.49%, representing a reduction of 6.51% in the death rate, through the formula applied to the treatment of poultry litter based on oxidising agents used as antiseptics, hydrogen peroxide solutions, glacial acetic acid and EDTA in order to act on bacteria, viruses, micro bacteria and spores.

Keywords: innovation, triple helix, poultry farms, biosecurity

Procedia PDF Downloads 269

Authors: Muna Jalal Ali, Essam A. Makky, Mashitah M. Yusoff

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Objectives: The aim of this study to evaluate the antimicrobial susceptibility of combined toothpaste with medicinal plants and the relations between the commercial toothpaste to its price and the patient age as well. Materials and Methods: Oral isolates of different patients aged 3 to 60 years were obtained, purified, and tested against four different ethnomedicinal plant extracts for antimicrobial activity. A total of 10 different commercial toothpastes (different brands and prices) were collected from the market, and the combined action of the medicinal plants and toothpaste was studied. Results: We found a higher bacterial population in the age group of 3–40 years than the group of 40–60 years, with approximately 44% and 32%, respectively. The combined action of ethanolic extract (alone) against oral isolates showed a synergistic effect, with 32.20, 30.50, and 25.42% for combinations A (Ci/Ca), B (Ci/Ca/P), and C (Ci/Ca/P/N), respectively. By contrast, the combined action of ethnomedicinal plants with 10 different toothpastes improved the antimicrobial sensitivity by 60, 100, and 0% for combinations A, B, and C respectively. Clinical relevance: The ethanolic extract of only combinations A and B with commercial toothpaste showed high antibacterial activity against oral isolates and the effectiveness of toothpaste is not related to the price.

Keywords: microbial evolution, oral isolates, ethnomedicinal plants, antimicrobial activity, toothpaste

Procedia PDF Downloads 291

Authors: Mohammed Jibrin Ndejiko

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Modern and current models such as flow cell technology which enhances a non-destructive growth and inspection of the sessile microbial communities revealed a great understanding of biofilms. A continuous flow system was designed to evaluate possibility of biofilm formation by Escherichia coli DH5α on the stainless steel (type 304) under continuous nutrient supply. The result of the colony forming unit (CFU) count shows that bacterial attachment and subsequent biofilm formation on stainless steel coupons with average surface roughness of 1.5 ± 1.8 µm and 2.0 ± 0.09 µm were both significantly higher (p ≤ 0.05) than those of the stainless steel coupon with lower surface roughness of 0.38 ± 1.5 µm. These observations support the hypothesis that surface profile is one of the factors that influence biofilm formation on stainless steel surfaces. The SEM and FESEM micrographs of the stainless steel coupons also revealed the attached Escherichia coli DH5α biofilm and dehydrated extracellular polymeric substance on the stainless steel surfaces. Thus, the fabricated flow system represented a very useful tool to study biofilm formation under continuous nutrient supply.

Keywords: biofilm, flowcell, stainless steel, coupon

Procedia PDF Downloads 305

Authors: Z. Herceg, V. Stulic, T. Vukusic, A. Rezek Jambrak

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High voltage electrical discharge plasmas are new nonthermal developing techniques used for water decontamination. To the full understanding of cell inactivation mechanisms, this study brings inactivation, recovery and cellular leakage of L. innocua cells before and after the treatment. Bacterial solution (200 mL) of L. innocua was treated in a glass reactor with a point-to-plate electrode configuration (high voltage electrode-titanium wire, was in the gas phase and grounded electrode was in the liquid phase). Argon was injected into the headspace of the reactor at the gas flow of 5 L/min. Frequency of 60, 90 and 120 Hz, time of 5 and 10 min, positive polarity and conductivity of media of 100 µS/cm were chosen to define listed parameters. With a longer treatment time inactivation was higher as well as the increase in cellular leakage. Despite total inactivation recovery of cells occurred probably because of a high leakage of proteins, compared to lower leakage of nucleic acids (DNA and RNA). In order to define mechanisms of inactivation further research is needed.

Keywords: Listeria innocua ATCC 33092, inactivation, gas phase plasma, cellular leakage, recovery of cells

Procedia PDF Downloads 159

Authors: R. Gounina-Allouane, I. Moussaoui, N. Boukahel

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Bacillus antimicrobial metabolites, especially those of Bacillus thuringiensis (Bt), are of great interest for research because of health risks generated by the excessive use of chemical additives as well as the propagation of resistant microbial strains, caused by the massive treatment with antibiotics. The objective of this study was the selection of Bt strains producing antimicrobial peptides (bacteriocins), and the partial purification of the most powerful bacteriocins, then the determination of their spectra of antimicrobial action. A collection of twenty one Bt strains isolated from soil at Boumerdès (northern Algeria) was used for screening strains having an antagonistic activity against phylogenetically closed bacteria. Spectra of antagonistic activity of two selected strains was determined against other Bt strains, Gram positive and Gram negative bacterial strains of clinical origin and others from ATCC collection as well as yeasts isolated in human dermatology. Bacteriocins of these two strains were partially purified and their effect on the kinetics of growth of the most sensitive microbial strains was studied. The bacteriocinogenic strains were biochemically characterized and their sensitivity to antibiotics was studied.

Keywords: antimicrobial peptides, Bacillus thuringiensis, bacteriocin, partial purification

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Authors: Suchao Donpudsa, Suwattana Visetnan, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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The Single Wap Domain (SWD) is a type III crustin antimicrobial peptide whose function is to defense the host animal against the bacterial infection by means of antimicrobial and antiproteinase activities. A study of LvSWD from Litopenaeus vannamei is reported herein about its activities and function against bacteria, particularly the Vibrio parahaemolyticus AHPND (VPAHPND) that causes acute hepatopancreatic necrosis disease. The over-expressed mature recombinant (r)LvSWD exhibits antimicrobial activity against both Gram-positive and Gram-negative bacteria, especially VPAHPND. With four times the MIC of rLvSWD, the treated post larval shrimp infected by VPAHPND is able to survive longer with the 50% survival rate as long as 78 h as compared to 36 h of the infected shrimp without rLvSWD. To a certain extent, we have demonstrated that the rLvSWD can be applied to protect the post larval shrimp.

Keywords: crustin, Litopenaeus vannamei, Vibrio parahaemolyticus AHPND, antimicrobial activity

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Authors: Jiri Nepereny, Vladimir Vrzal

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Staphylococcus intermedius/pseudintermedius bacteria are commonly found on the skin of healthy dogs and can cause pruritic skin diseases under certain circumstances (trauma, allergy, immunodeficiency, ectoparasitosis, endocrinological diseases, glucocorticoid therapy, etc.). These can develop into complicated superficial or deep pyoderma, which represent a large group of problematic skin diseases in dogs. These are predominantly inflammations of a secondary nature, associated with the occurrence of coagulase-positive Staphylococcus spp. A major problem is increased itching, which greatly complicates the healing process. The aim of this work is to verify the efficacy of the developed preparation Bacteriophage SI (Staphylococcus intermedius). The tested preparation contains a lysate of bacterial cells of S. intermedius host culture including culture medium and live virions of specific phage. Sodium Merthiolate is added as a preservative in a safe concentration. Validation of the efficacy of the product was demonstrated by monitoring the therapeutic effect after application to indicated cases from clinical practice. The indication for inclusion of the patient into the trial was an adequate history and clinical examination accompanied by sample collection for bacteriological examination and isolation of the specific causative agent. Isolate identification was performed by API BioMérieux identification system (API ID 32 STAPH) and rep-PCR typing. The suitability of therapy for a specific case was confirmed by in vitro testing of the lytic ability of the bacteriophage to lyse the specific isolate = formation of specific plaques on the culture isolate on the surface of the solid culture medium. So far, a total of 32 dogs of different sexes, ages and breed affiliations with different symptoms of staphylococcal dermatitis have been included in the testing. Their previous therapy consisted of more or less successful systemic or local application of broad-spectrum antibiotics. The presence of S. intermedius/pseudintermedius has been demonstrated in 26 cases. The isolates were identified as a S. pseudintermedius, in all cases. Contaminant bacterial microflora was always present in the examined samples. The test product was applied subcutaneously in gradually increasing doses over a period of 1 month. After improvement in health status, maintenance therapy was followed by application of the product once a week for 3 months. Adverse effects associated with the administration of the product (swelling at the site of application) occurred in only 2 cases. In all cases, there was a significant reduction in clinical signs (healing of skin lesions and reduction of inflammation) after therapy and an improvement in the well-being of the treated animals. A major problem in the treatment of pyoderma is the frequent resistance of the causative agents to antibiotics, especially the increasing frequency of multidrug-resistant and methicillin-resistant S. pseudintermedius (MRSP) strains. Specific phagolysate using for the therapy of these diseases could solve this problem and to some extent replace or reduce the use of antibiotics, whose frequent and widespread application often leads to the emergence of resistance. The advantage of the therapeutic use of bacteriophages is their bactericidal effect, high specificity and safety. This work was supported by Project FV40213 from Ministry of Industry and Trade, Czech Republic.

Keywords: bacteriophage, pyoderma, staphylococcus spp, therapy

Procedia PDF Downloads 157

Authors: Mosaab A. Omar, Mohammad Saleh Al-Aboody, Mohmed A. Rizk, Shimaa M. Elsayed, Ahmed ElSify, Naoaki Yokoyama, Ikuo Igarashi

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Enoxacin is a broad-spectrum 6-fluoronaphthyridinone antibacterial agent (fluoroquinolones) structurally related to nalidixic acid used mainly in the treatment of urinary tract infections and gonorrhea. Also, it has been shown recently that it may have cancer inhibiting effect. The primary antibabesial effect of Enoxacin is due to inhibition of DNA gyrase subunit A, and DNA topoisomerase. In the present study, enoxacin was tested as a potent inhibitor against the in vitro growth of bovine and equine Piroplasms. The in vitro growth of five Babesia species that were tested was significantly inhibited (P<0.05) by micromolar concentrations of enoxacin (IC50 values= 13.5, 7.2, 7.5, and 24.2 µM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Enoxacin IC50 values for Babesia and Theileria parasites were satisfactory as the drug is a potent antibacterial drug with minimum side effects. Therefore, enoxacin might be used for the treatment of Babesiosis and Theileriosis especially in case of mixed infections with bacterial diseases or in the case of animal sensitivity against diminazin toxicity.

Keywords: enoxacin, Babesia, Theileria, IC50 and dimenazin

Procedia PDF Downloads 451

Authors: David D. Adams, Ibrahim B. Bello

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Spent oil contaminated Soil from ten selected mechanic workshops were investigated for their bacteria and bioremediation potentials. The bacterial isolates were morphologically and molecularly identified as Enterobacter hormaechei, Escherichia coli, Klebsiella pneumoniae, Shigella flexneri , Wesiella cibaria, Lactobacillus planetarium. The singles and a consortium of these bacteria incubated in the minimal salt medium incorporated with 1% engine oil exhibited various biodegradation rates, with the mixed consortium exhibiting the highest for this oil. The gene for the hydrocarbon enzyme Catechol 2, 3 dioxygenase (C2,30) was detected and amplified in Enterobacter hormaechei, Escherichia coli and Shigella flexneri using PCR and Agarose gel electrophoresis. The detection of the (C2,30) enzyme gene in, and the spent oil biodegradation activity exhibited by these bacteria suggest their possible possession of bioremediating potentials for the spent engine oil. It is therefore suggested that a pilot study on the field application of these bacteria for bioremediation and restoration of spent oil polluted environment should be done in mechanic workshops.

Keywords: spent engine oil, pollution, bacteria, enzyme, bioremediation, mechanic workshop

Procedia PDF Downloads 200

Authors: R. Gounina-Allouane, I. Moussaoui, N. Boukahel

Abstract:

Bacillus antimicrobial metabolites, especially those of Bacillus thuringiensis (Bt), are of great interest for research because of health risks generated by the excessive use of chemical additives as well as the propagation of resistant microbial strains, caused by the massive treatment with antibiotics. The objective of this study was the selection of Bt strains producing antimicrobial peptides (bacteriocins), and the partial purification of the most powerful bacteriocins, then the determination of their spectra of antimicrobial action. A collection of twenty one Bt strains isolated from soil at Boumerdès (northern of Algeria) was used for screening strains having an antagonistic activity against phylogenetically closed bacteria. Spectra of antagonistic activity of two selected strains was determined against other Bt strains, Gram positive and Gram negative bacterial strains of clinical origin and others from ATCC collection as well as yeasts isolated in human dermatology. Bacteriocins of these two strains were partially purified and their effect on the kinetics of growth of the most sensitive microbial strains was studied. The bacteriocinogenic strains were biochemically characterized and their sensitivity to antibiotics was studied.

Keywords: antimicrobial peptides, Bacillus thuringiensis, bacteriocin, partial purification

Procedia PDF Downloads 344

Authors: Khaoula Bensaida, Osama Eljamal

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The electrical energy generation through Microbial Fuel Cells (MFCs) using microorganisms is a renewable and sustainable approach. It creates truly an efficient technology for power production and wastewater treatment. MFC is an electrochemical device which turns wastewater into electricity. The most important part of MFC is microbes. Nano zero-valent Iron NZVI technique was successfully applied in degrading the chemical pollutants and cleaning wastewater. However, the use of NZVI for enhancing the current production is still not confirmed yet. This study aims to confirm the effect of these particles on the current generation by using MFC. A constructed microbial fuel cell, which utilizes domestic wastewater, has been considered for wastewater treatment and bio-electricity generation. The two electrodes were connected to an external resistor (200 ohms). Experiments were conducted in two steps. First, the MFC was constructed without adding NZVI particles (Control) while at a second step, nanoparticles were added with a concentration of 50mg/L. After 20 hours, the measured voltage increased to 5 and 8mV, respectively. To conclude, the use of zero-valent iron in an MFC system can increase electricity generation.

Keywords: bacterial growth, electricity generation, microbial fuel cell MFC, nano zero-valent iron NZVI.

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Authors: Prajna Metta, Yanti Rusyanti, Nunung Rusminah, Bremmy Laksono

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The decrease of neutrophil chemotaxis function may cause increased susceptibility to aggressive periodontitis (AP). Neutrophil chemotaxis is affected by formyl peptide receptor 1 (FPR1), which when activated will respond to bacterial chemotactic peptide formyl methionyl leusyl phenylalanine (FMLP). FPR1 protein value is decreased in response to a wide number of inflammatory stimuli in AP patients. This study was aimed to assess the alteration of FPR1 protein value in AP patients and if FPR1 protein value could be used as an indicator of neutrophil chemotaxis dysfunction in AP. This is a case control study with 20 AP patients and 20 control subjects. Three milliliters of peripheral blood were drawn and analyzed for FPR1 protein value with ELISA. The data were statistically analyzed with Mann-Whitney test (p>0,05). Results showed that the mean value of FPR1 protein value in AP group is 0,353 pg/mL (0,11 to 1,18 pg/mL) and the mean value of FPR1 protein value in control group is 0,296 pg/mL (0,05 to 0,88 pg/mL). P value 0,787 > 0,05 suggested that there is no significant difference of FPR1 protein value in both groups. The present study suggests that FPR1 protein value has no significance alteration in AP patients and could not be used as an indicator of neutrophil chemotaxis dysfunction.

Keywords: aggressive periodontitis, chemotaxis dysfunction, FPR1 protein value, neutrophil

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Authors: Mona Hajirasouli

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Abstract: introduction: Antimicrobial compounds are important in the first line of the host defense system of many animal species. Material and methods: In the present study antibacterial activity of crude and proteins precipitate of hemolymph and crude hepatopancreas extracts from Portunus segnis and Grapsus albolineatus against a range of 6 different bacterial strains evaluated. Amoxicillin as a positive control were also used. Results: Maximum activity (15.9 mm) was recorded in male haemolymph of p.segnis against Entrobacter and minimum activity (7 mm) was recorded against Serratia marcescens, Enterobacter sp. and Proteus mirabilis from different extracts of Grapsus albolineatus. Data were analyzed using independent-t in SPSS version 16, and results indicate that there were not any significant differences between hemolymph and hepatopancreas extracts of 2 species. Discussion: Antimicrobial activity has been reported earlier in the hemolymph of some brachyuran crabs such as: blue crab Callinectes sapidus, mud crab Scylla serrata, Ocypode macrocera and Carcinus maenas. This study shows that hemolymph and hepatopancreas of Portunus segnis and Grapsus albolineatus may potential antibiotics.

Keywords: brachyuran, Portunus segnis, Grapsus albolineatus, hemolymph, hepatopancreas, antibacterial

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Authors: Ikram Kamal, Mohamed Blaghen

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Pollution caused by petroleum hydrocarbons in terrestrial and aquatic environment is a common phenomenon that causes significant ecological and social problems. Biosurfactant applications in the environmental industries are promising due to their biodegradability, low toxicity and effectiveness in enhancing biodegradation and solubilization of low solubility compounds. Currently, the main application is for enhancement of oil recovery and hydrocarbon bioremediation due to their biodegradability and low critical micelle concentration (CMC). In this study we have investigated the potential of bacterial strains collected aseptically from the lagoon Marchika (water and soil) in Nador, Morocco; for the production of biosurfactants. This study also aimed to optimize the biosurfactant production process by changing the variables that influence the type and amount of biosurfactant produced by these microorganisms such as: carbon sources and also other physical and chemical parameters such as temperature and pH. Emulsification index, methylene blue test and thin layer chromatography (TLC) revealed the ability of strains used in this study to produce compounds that could emulsify gasoline. In addition a GC/MS was used to separate and identify different biosurfactants purified.

Keywords: petroleum hydrocarbons, biosurfactant, biodegradability, critical micelle concentration, lagoon Marchika

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Authors: Austin Belfiori, Kevin Rinek, Zach Barcroft, Jennifer Berglind

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Diet influences the composition of the gut microbiota and host's health. Disruption of the balance among the microbiota, epithelial cells, and resident immune cells in the intestine is involved in the pathogenesis of inflammatory bowel disease (IBD). To study the role of gut microbiota in intestinal inflammation, the microbiome of control mice (C57BL6) given a gluten-containing standard diet versus C57BL6 mice given the gluten-free (GF) feed (n=10 in each group) was examined. All mice received the 3% DSS for 5 days. Throughout the study, feces were collected and processed for DNA extraction and MiSeq Illumina sequencing of V4 region of bacterial 16S rRNA gene. Alpha and beta diversities and compositional differences at phylum and genus levels were determined in intestinal microbiota. The mice receiving the GF diet showed a significantly increased abundance of Firmicutes and a decrease of Bacteroides and Lactobacillus at phylum level. Therefore, the gluten free diet led to reductions in beneficial gut bacteria populations. These findings indicate a role of wheat gluten in dysbiosis of the intestinal microbiota.

Keywords: gluten, colitis, microbiota, DSS, dextran sulphate sodium

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Authors: Tahisa M. Pedroso, Hérida R. N. Salgado

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The microbiological turbidimetric assay allows the determination of potency of the drug, by measuring the turbidity (absorbance), caused by inhibition of microorganisms by ertapenem sodium. Ertapenem sodium (ERTM), a synthetic antimicrobial agent of the class of carbapenems, shows action against Gram-negative, Gram-positive, aerobic and anaerobic microorganisms. Turbidimetric assays are described in the literature for some antibiotics, but this method is not described for ertapenem. The objective of the present study was to develop and validate a simple, sensitive, precise and accurate microbiological assay by turbidimetry to quantify ertapenem sodium injectable as an alternative to the physicochemical methods described in the literature. Several preliminary tests were performed to choose the following parameters: Staphylococcus aureus ATCC 25923, IAL 1851, 8 % of inoculum, BHI culture medium, and aqueous solution of ertapenem sodium. 10.0 mL of sterile BHI culture medium were distributed in 20 tubes. 0.2 mL of solutions (standard and test), were added in tube, respectively S1, S2 and S3, and T1, T2 and T3, 0.8 mL of culture medium inoculated were transferred to each tube, according parallel lines 3 x 3 test. The tubes were incubated in shaker Marconi MA 420 at a temperature of 35.0 °C ± 2.0 °C for 4 hours. After this period, the growth of microorganisms was inhibited by addition of 0.5 mL of 12% formaldehyde solution in each tube. The absorbance was determined in Quimis Q-798DRM spectrophotometer at a wavelength of 530 nm. An analytical curve was constructed to obtain the equation of the line by the least-squares method and the linearity and parallelism was detected by ANOVA. The specificity of the method was proven by comparing the response obtained for the standard and the finished product. The precision was checked by testing the determination of ertapenem sodium in three days. The accuracy was determined by recovery test. The robustness was determined by comparing the results obtained by varying wavelength, brand of culture medium and volume of culture medium in the tubes. Statistical analysis showed that there is no deviation from linearity in the analytical curves of standard and test samples. The correlation coefficients were 0.9996 and 0.9998 for the standard and test samples, respectively. The specificity was confirmed by comparing the absorbance of the reference substance and test samples. The values obtained for intraday, interday and between analyst precision were 1.25%; 0.26%, 0.15% respectively. The amount of ertapenem sodium present in the samples analyzed, 99.87%, is consistent. The accuracy was proven by the recovery test, with value of 98.20%. The parameters varied did not affect the analysis of ertapenem sodium, confirming the robustness of this method. The turbidimetric assay is more versatile, faster and easier to apply than agar diffusion assay. The method is simple, rapid and accurate and can be used in routine analysis of quality control of formulations containing ertapenem sodium.

Keywords: ertapenem sodium, turbidimetric assay, quality control, validation

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Authors: E. Kuley, M. Durmuş, E. Balikci, G. Ozyurt, Y. Uçar, F. Kuley, F. Ozogul, Y. Ozogul

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Ammonia and biogenic amine production of fish spoilage bacteria in sardine infusion decarboxylase broth and antimicrobial effect of olive by products (olive leaf extract:OL, olive cake: OC and black water:BW) was monitored using HPLC method. Fish spoilage bacteria produced all biogenic amine tested, mainly histamine and serotonin. Ammonia was accumulated more than 13.60 mg/L. Histamine production was in range 37.50 mg/L by Ser. liquefaciens and 86.71 mg/L by Ent. cloacae. The highest putrescine and cadaverine production was observed by Ent. cloacae (17.80 vs. 17.69 mg/L). The presence of OL, OC and BW in the broth significantly affected biogenic amine accumulation by bacteria. The antibacterial effect of olive by products depended on bacterial strains. OL and OC resulted in significant inhibition effect on HIS accumulation by bacteria apart from Ser. liquefaciens and Prot. mirabilis. The study result revealed that usefulness of OL and OC to prevent the accumulation of this amine which may affect human health.

Keywords: Antimicrobials, biogenic amine, fish spoilage bacteria, olive-by products

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Authors: A. Sebti Bouzid, S. Igoud, L. Aoudjit, H. Lebik

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Mathematical modeling and numerical simulation have emerged over the past two decades as one of the key tools for design and optimize performances of physical and chemical processes intended to water disinfection. Water photolysis is an efficient and economical technique to reduce bacterial contamination. It exploits the germicidal effect of solar ultraviolet irradiation to inactivate pathogenic microorganisms. The design of photo-reactor operating in continuous disinfection system, required tacking in account the hydrodynamic behavior of water in the reactor. Since the kinetic of disinfection depends on irradiation intensity distribution, coupling the hydrodynamic and solar radiation distribution is of crucial importance. In this work we propose a numerical simulation study for hydrodynamic and solar irradiation distribution in a tubular photo-reactor. We have used the Computational Fluid Dynamic code Fluent under the assumption of three-dimensional incompressible flow in unsteady turbulent regimes. The results of simulation concerned radiation, temperature and velocity fields are discussed and the effect of inclination angle of reactor relative to the horizontal is investigated.

Keywords: solar water disinfection, hydrodynamic modeling, solar irradiation modeling, CFD Fluent

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Authors: Amanda Gregorim Fernandes, Lorena Cardoso Cintra, Rosalia Santos Amorim Jesuino, Fabricia Paula De Faria, Marcio José Poças Fonseca

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Bioethanol is one of the most promising biofuels and able to replace fossil fuels and reduce its different environmental impacts and can be generated from various agroindustrial waste. The Brazil is in first place in bioethanol production to be the largest producer of sugarcane. The bagasse sugarcane (SCB) has lignocellulose which is composed of three major components: cellulose, hemicellulose and lignin. Cellulose is a homopolymer of glucose units connected by glycosidic linkages. Among all species of Penicillium, Penicillium echinulatum has been the focus of attention because they produce high quantities of cellulase and the mutant strain 9A02S1 produces higher enzyme levels compared to the wild. Among the cellulases, the cellobiohydrolases enzymes are the main components of the cellulolytic system of fungi, and are also responsible for most of the potential hydrolytic in enzyme cocktails for the industrial processing of plant biomass and several cellobiohydrolases Penicillium had higher specific activity against cellulose compared to CBH I from Trichoderma reesei. This fact makes it an interesting pattern for higher yields in the enzymatic hydrolysis, and also they are important enzymes in the hydrolysis of crystalline regions of cellulose. Therefore, finding new and more active enzymes become necessary. Meanwhile, β-glycosidases act on soluble substrates and are highly dependent on cellobiohydrolases and endoglucanases action to provide the substrate in the hydrolysis of the biomass, but the cellobiohydrolases and endoglucanases are highly dependent β-glucosidases to maintain efficient hydrolysis. Thus, there is a need to understand the structure-function relationships that govern the catalytic activity of cellulolytic enzymes to elucidate its mechanism of action and optimize its potential as industrial biocatalysts. To evaluate the enzyme β-glucosidase of Penicillium echinulatum (PeBGL1) the gene was synthesized from the assembly sequence from a library in induction conditions and then the PeBGL1 gene was cloned in the vector pPICZαA and transformed into P. pastoris GS115. After processing, the producers of PeBGL1 were analyzed for enzyme activity and protein profile where a band of approximately 100 kDa was viewed. It was also carried out the zymogram. In partial characterization it was determined optimum temperature of 50°C and optimum pH of 6,5. In addition, to increase the secreted recombinant PeBGL1 production by Pichia pastoris, three parameters of P. pastoris culture medium were analysed: methanol, nitrogen source concentrations and the inoculum size. A 23 factorial design was effective in achieving the optimum condition. Altogether, these results point to the potential application of this P. echinulatum β-glucosidase in hydrolysis of cellulose for the production of bioethanol.

Keywords: bioethanol, biotechnology, beta-glucosidase, penicillium echinulatum

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Authors: Andrew Read, Alice Parry, Kate Woods

Abstract:

Since the advent of the SARS-CoV-2 pandemic, the search for medications that can reduce mortality and morbidity has been a global research priority. Several multi-center trials have recently demonstrated improved mortality associated with the use of Tocilizumab, an interleukin-6 receptor antagonist, in patients with severe SARS-CoV-2 pneumonia. Initial data supported the administration in patients requiring respiratory support (non-invasive or invasive ventilation), but more recent data has shown benefit in all hypoxic patients. At the height of the second wave of COVID-19 infections in London, our hospital introduced the use of Tocilizumab for patients with severe COVID-19. Tocilizumab is licensed for use in chronic inflammatory conditions and has been associated with an increased risk of severe bacterial and fungal infections, as well as reactivation of chronic viral infections (e.g., hepatitis B). It is a specialist drug that suppresses the formation of C-reactive protein (CRP) for 6 – 12 weeks. It is not widely used by the general medical community. We aimed to assess Tocilizumab use in our hospital and to implement changes to the protocol as required to ensure administration was safe and appropriate. A retrospective study design was used to assess prescriptions over an initial 3-week period in both intensive care and on the medical wards. This amounted to a total of 13 patients. The initial data collection identified four key areas of concern: adherence to national and local inclusion & exclusion criteria; a collection of appropriate screening blood prior to administration; documentation of informed consent or best interest decision and documentation of Tocilizumab administration on patient discharge information, to alert future healthcare providers that typical measures of inflammation and infection, such as CRP, are unreliable for up to 3-months. Data were collected from electronic notes, blood results and observation charts, and cross referenced with pharmacy data. Initial results showed that all four key areas were completed in approximately 50% of cases. Of particular concern was adherence to exclusion criteria, such as current evidence of bacterial infection, and ensuring the correct screening blood was sent to exclude infections such as hepatitis. To remedy this and improve patient safety, the initial data was presented to relevant healthcare professionals. Subsequently, three interventions were introduced and education on each provided to hospital staff. An electronic ‘order set’ collating the appropriate screening blood was created simplifying the screening process. Pre-formed electronic documentation which can be inserted into the notes was created to provide a framework for consent discussions and reduce the time needed for junior doctors to complete this task. Additionally, a ‘Tocilizumab’ administration card was created and administered via pharmacy. This was distributed to each patient on discharge to ensure future healthcare professionals were aware of the potential effects of Tocilizumab administration, including suppression of CRP. Following these changes, repeat data collection over two months illustrated that each of the 4 safety aspects was met with a 100% success rate in every patient. Although this demonstrates good progress and effective interventions the challenge will be to maintain this progress. The audit data collection is ongoing

Keywords: education, patient safety , SARS-CoV-2, Tocilizumab

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Authors: Geeta Sharma, Suma Chandra

Abstract:

Sugarcane is one of the major commercial crop playing roles in agriculture and industrial economy of India. Globally sugarcane is affected by approximately 240 diseases caused by various plant pathogenic organisms. Among them, red rot disease caused by the fungus Colletotrichum falcatum, is one of the most important diseases. In the present investigation, one fungal bioagent of Trichoderma harzianum, Pant Bioagent 1 and one bacterial bioagent Pseudomonas fluorescence, Pant Bioagent 2 (PBAT 1 and PBAT 2, respectively) were tested by dual culture method against the pathogen under laboratory conditions. The effectiveness of biocontrol agents was observed against four isolates of C. falcatum. In the case of PBAT1 maximum percent inhibition of pathogen was recorded in isolated Cf 0238 (61.05%), followed by Cf 09 (60.62%) whereas, minimum percent inhibition was recorded in Cf 3220 (48.55%) and in case of PBAT2 maximum mycelial growth inhibition percent was recorded in Cf 767 (50.50%) followed by Cf 088230(48.83%), whereas minimum percent inhibition was recorded in Cf 08 (40.16%) followed by Cf 0238 (41.83%). The present study showed that these biocontrol agents have the potential of controlling the pathogen and can further be used for the management of red rot disease in field.

Keywords: biocontrol agents, Colletotrichum falcatum, isolates, sugarcane

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Authors: Neda Sinaei, Davood Zare, Mehrdad Azin

Abstract:

Polyhydroxyalkanoates (PHAs) are biocompatible and biodegradable polymers produced by a wide range of bacterial strains. These biopolymers are significantly studied for drug delivery and tissue engineering applications because of their fascinating physicochemical properties. Polyhydroxybutyrate (PHB) scaffold that has been extracted from a novel bacteria using oil wastewater was selected to study. Some physical parameters affecting scaffold properties such as PHB concentration, solvent evaporation speed, and ultrasonic time were investigated. Scanning electron microscopy was used to evaluate the porosity. Afterward, the biocompatibility of PHB scaffold was assessed. Initial results showed the highly porous PHB scaffold structure with a variety of pore sizes. Subsequent results indicated that more unique pore sizes can be obtained by optimizing physical factors. It would be noticed that the morphology of the pore structure was accordingly affected by ultrasonic time. Hence, In vitro cell viability tests on the PHB scaffold using human foreskin fibroblasts revealed strong cell attachment and proliferation supports. Therefore, it can be concluded that the cost-effective PHB scaffold has the potential using as a biomaterial cell adhesion substrate in therapeutic applications.

Keywords: Polyhydroxybutyrate, biocompatible, scaffold, porous, tissue engineering

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Authors: L. Rakesh, T. Y. Heblekar

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This study entails the development and optimization of a mathematical model for a floating drum biogas reactor from first principles using thermal and empirical considerations. The model was derived on the basis of mass conservation, lumped mass heat transfer formulations and empirical biogas formation laws. The treatment leads to a system of coupled nonlinear ordinary differential equations whose solution mapped four-time independent controllable parameters to five output variables which adequately serve to describe the reactor performance. These equations were solved numerically using fourth order Runge-Kutta method for a range of input parameter values. Using the data so obtained an Artificial Neural Network with a single hidden layer was trained using Levenberg-Marquardt Damped Least Squares (DLS) algorithm. This network was then fine-tuned for optimal mapping by varying hidden layer size. This fast forward model was then employed as a health score generator in the Bacterial Foraging Optimization code. The optimal operating state of the simplified Biogas reactor was thus obtained.

Keywords: biogas, floating drum reactor, neural network model, optimization

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Authors: Elmira Davasaz Tabrizi, Müşteba Sevil, Ercan Arican

Abstract:

In recent decades, there has been a sharp increase in the prevalence of several unrelated chronic diseases. The use of long-term antibiotics for chronic illnesses is increasing. The antibiotic resistance occurrence and its relationship with host microbiomes are still unclear. Properties of the identifying antibodies have been the focus of chronic disease research, such as prostatitis or autoimmune. The immune system is made up of a complicated but well-organized network of cell types that constantly monitor and maintain their surroundings. The regulated homeostatic interaction between immune system cells and their surrounding environment shapes the microbial flora. Researchers believe that the disappearance of special bacterial species from our ancestral microbiota might have altered the body flora that can cause a rise in disease during the human life span. This unpleasant pattern demonstrates the importance of focusing on discovering and revealing the root causes behind the disappearance or alteration of our microbiota. In this review, we gathered the results of some studies that reveal changes in the diversity and quantity of microorganisms that may affect chronic and autoimmune diseases. Additionally, a Ph.D. thesis that is still in process as Metagenomic studies in chronic prostatitis samples is mentioned.

Keywords: metagenomic, autoimmune, prostatitis, microbiome

Procedia PDF Downloads 80

Authors: P. Serrano-Pérez, M. C. Rodríguez-Molina, C. Palo, E. Palo, A. Lacasa

Abstract:

Phytophthora nicotianae is the principal causal agent of root and crown rot disease of red pepper plants in Extremadura (Western Spain). There is a need to develop a biologically-based method of soil disinfestation that facilitates profitable and sustainable production without the use of chemical fumigants. Anaerobic Soil Disinfestation (ASD), as well know as biodisinfestation, has been shown to control a wide range of soil-borne pathogens and nematodes in numerous crop production systems. This method implies soil wetting, incorporation of a easily decomposable carbon-rich organic amendment and covering with plastic film for several weeks. ASD with rapeseed cake (var. Tocatta, a glucosinolates-free variety) used as C-source was assayed in spring 2014, before the pepper crop establishment. The field experiment was conducted at the Agricultural Research Centre Finca La Orden (Southwestern Spain) and the treatments were: rapeseed cake (RCP); rapeseed cake without plastic cover (RC); control non-amendment (CP) and control non-amendment without plastic cover (C). The experimental design was a randomized complete block design with four replicates and a plot size of 5 x 5 m. On 26 March, rapeseed cake (1 kg·m-2) was incorporated into the soil with a rotovator. Biological probes with the inoculum were buried at 15 and 30-cm depth (biological probes were previously prepared with 100 g of disinfected soil inoculated with chlamydospores (chlam) of P. nicotianae P13 isolate [100 chlam·g-1 of soil] and wrapped in agryl cloth). Sprinkler irrigation was run until field capacity and the corresponding plots were covered with transparent plastic (PE 0.05 mm). On 6 May plastics were removed, the biological probes were dug out and a bioassay was established. One pepper seedling at the 2 to 4 true-leaves stage was transplanted in the soil from each biological probe. Plants were grown in a climatic chamber and disease symptoms were recorded every week during 2 months. Fragments of roots and crown of symptomatic plants were analyzed on NARPH media and soil from rizospheres was analyzed using carnation petals as baits. Results of “survival” were expressed as the percentage of soil samples where P. nicotianae was detected and results of “infectivity” were expressed as the percentage of diseased plants. No differences were detected in deep effect. Infectivity of P. nicotianae chlamydospores was successfully reduced in RCP treatment (4.2% of infectivity) compared with the controls (41.7% of infectivity). The pattern of survival was similar to infectivity observed by the bioassay: 21% of survival in RCP; 79% in CP; 83% in C and 87% in RC. Although ASD may be an effective alternative to chemical fumigants to pest management, more research is necessary to show their impact on the microbial community and chemistry of the soil.

Keywords: biodisinfestation, BSD, soil fumigant alternatives, organic amendments

Procedia PDF Downloads 200

Authors: Sunil Kumar

Abstract:

For many years, small organic molecules derived naturally from microbes and plants have delivered a number of expedient therapeutic drug agents. The search for naturally occurring lead compounds has continued in recent years as well, with the constituents of marine flora and fauna along with those of telluric microorganisms and plants being investigated for their anti-bacterial and anti-cancer activities. It has been observed that such promising lead molecules incline to promptly generate substantial attention among scientists like synthetic organic chemists and biologists. Subsequently, the availability of a given precious natural product sample may be enriched, and it may be possible to determine a preliminary idea of structure-activity relationships to develop synthetic analogues. For instance, anti-tumor drug topotecan is a synthetic chemical compound similar in chemical structure to camptothecin which is found in extracts of Camptotheca acuminate. Similarly, researchers at AstraZeneca discovered anti-biotic pyrrolamide through a fragment-based lead generation approach from kibdelomycin, which is isolated from Staphylococcus aureuss.

Keywords: anticancer, antibiotic, lead molecule, natural product, synthetic analogues

Procedia PDF Downloads 134