Search results for: principal strains
1398 Role of ABC-Type Efflux Transporters in Antifungal Resistance of Candida auris
Authors: Mohamed Mahdi Alshahni, Takashi Tamura, Koichi Makimura
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Objective: The objective of this study is to evaluate roles of ABC-type efflux transporters in the resistance of Candida auris against common antifungal agents. Material and Methods: A wild-type C. auris strain and its antifungal resistant derivative strain that is generated through induction by antifungal agents were used in this study. The strains were cultured onto media containing beauvericin alone or in combination with azole agents. Moreover, expression levels of four ABC-type transporter’s homologs in those strains were analyzed by real time PCR with or without antifungal stress by fluconazole or voriconazole. Results: Addition of beauvericin helped to partially restore the susceptibility of the resistant strain against fluconazole, suggesting participation of ABC-type transporters in the resistance mechanism. Real time PCR results showed that mRNA levels of three out of the four analyzed transporters in the resistant strain were more than 2-fold higher than their counterparts in the wild-type strain under negative control and antifungal agent-containing conditions. Conclusion: C. auris is an emerging multidrug-resistant pathogen causing human mortality worldwide. Providing effective treatment has been hampered by the resistance to antifungal drugs, demanding understanding the resistance mechanism in order to devise new therapeutic strategies. Our data suggest a partial contribution of ABC-type transporters to the resistance of this pathogen.Keywords: resistance, C. auris, transporters, antifungi
Procedia PDF Downloads 1611397 Phytochemical Study and Antimicrobial Activity of Nigella sativa L. (Renunculaceae) in Algeria
Authors: L. Bendifallah, F. Acheuk, M. Djouabi, M. Oukili, R. Ghezraoui, W. Lakhdari, R. Allouane
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Nigella sativa L. (Renunculaceae) native to the Mediterranean region and Western Asia, Black cumin is grown to India, through Sudan and Ethiopia. It is widely cultivated in Egypt, the Middle East, Saudi Arabia, Turkey, Sudan, Afghanistan and Europe. It is among the most important medicinal plants in Algeria that is known for its antifungal and antimicrobial properties. Despite its plethora of uses for treating various diseases, it has garnered very little scientific interest so far, particularly in Algeria. For this study, the seeds of Algerian Nigella sativa L cultivated in the area of Magra (M’sila) in northern Algeria, were collected in summer. In such a propitious context, the aim of this study was to enhance Nigella sativa as a medicinal herb. The phytochemical screening methods are used. For their antimicrobial activity, extracts of tannin and polyphenols were screened against four pathogenic bacterial strains and two pathogenic yeast strains. The phytochemical analysis results showed a remarkable combination of chemical components including a high content in tannins, in flavonoïds, and in alkaloids. The tannins and the polyphenols have strong antimicrobial activity against all the species. The maximum zone of inhibition was noted for polyphenol and tannin extracts against Escerichia coli (14 mm, 12.33 mm) and an antifungic activity against Aspergillus niger (11.66 mm, 9 mm). These results indicate to some benefits of Nigella sativa seeds which can use to treatment the microbial infection.Keywords: Nigella sativa, phytochemistry, antimicrobial activity, Algeria
Procedia PDF Downloads 3181396 Phytochemical Study and Antimicrobial Activity of Nigella Sativa L. (Renunculaceae) in Algeria
Authors: L. Bendifallah, F.Acheuk, M. Djouabi, M. Oukili, R. Ghezraoui, W. Lakhdari, R. Allouane
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Nigella sativa L. (Renunculaceae) native to the Mediterranean region and Western Asia, Black cumin is grown to India, through Sudan and Ethiopia. It is widely cultivated in Egypt, the Middle East, Saudi Arabia, Turkey, Sudan, Afghanistan and Europe. It is among the most important medicinal plants in Algeria that is known for its antifungal and antimicrobial properties. Despite its plethora of uses for treating various diseases, it has garnered very little scientific interest so far, particularly in Algeria. For this study, the seeds of Algerian Nigella sativa L cultivated in the area of Magra (M’sila) in northern Algeria, were collected in summer. In such a propitious context, the aim of this study was to enhance Nigella sativa as a medicinal herb. The phytochemical screening methods are used. For their antimicrobial activity, extracts of tannin and polyphenols were screened against four pathogenic bacterial strains and two pathogenic yeast strains. The phytochemical analysis results showed a remarkable combination of chemical components including a high content in tannins, in flavonoïds, and in alkaloids. The tannins and the polyphenols have strong antimicrobial activity against all the species. The maximum zone of inhibition was noted for polyphenol and tannin extracts against Escerichia coli (14 mm, 12.33 mm) and an antifungic activity against Aspergillus niger (11.66 mm, 9 mm). These results indicate to some benefits of Nigella sativa seeds which can use to treatment the microbial infection.Keywords: Algeria, antimicrobial activity, Nigella sativa, phytochemistry
Procedia PDF Downloads 5591395 Numerical Simulations on the Torsional Behavior of Multistory Concrete Masonry Buildings
Authors: Alvaro Jose Cordova, Hsuan Teh Hu
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The use of concrete masonry constructions in developing countries has become very frequent, especially for domestic purpose. Most of them with asymmetric wall configurations in plan resulting in significant torsional actions when subjected to seismic loads. The study consisted on the finding of a material model for hollow unreinforced concrete masonry and a validation with experimental data found in literature. Numerical simulations were performed to 20 buildings with variations in wall distributions and heights. Results were analyzed by inspection and with a non-linear static method. The findings revealed that eccentricities as well as structure rigidities have a strong influence on the overall response of concrete masonry buildings. In addition, slab rotations depicted more accurate information about the torsional behavior than maximum versus average displacement ratios. The failure modes in low buildings were characterized by high tensile strains in the first floor. Whereas in tall buildings these strains were lowered significantly by higher compression stresses due to a higher self-weight. These tall buildings developed multiple plastic hinges along the height. Finally, the non-linear static analysis exposed a brittle response for all masonry assemblies. This type of behavior is undesired in any construction and the need for a material model for reinforced masonry is pointed out.Keywords: concrete damaged plasticity, concrete masonry, macro-modeling, nonlinear static analysis, torsional capacity
Procedia PDF Downloads 2871394 Comparative Growth Kinetic Studies of Two Strains Saccharomyces cerevisiae Isolated from Dates and a Commercial Strain
Authors: Nizar Chaira
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Dates, main products of the oases, due to their therapeutic interests, are considered highly nutritious fruit. Several studies on the valuation biotechnology and technology of dates are made, and several products are already prepared. Isolation of the yeast Saccharomyces cerevisiae, naturally presents in a scrap of date, optimization of growth in the medium based on date syrup and production biomass can potentially expand the range of secondary products of dates. To this end, this paper tries to study the suitability for processing dates technology and biotechnology to use the date pulp as a carbon source for biological transformation. Two strains of Saccharomyces cerevisiae isolated from date syrup (S1, S2) and a commercial strain have used for this study. After optimization of culture conditions, production in a fermenter on two different media (date syrup and beet molasses) was performed. This is followed by studying the kinetics of growth, protein production and consumption of sugars in crops strain 1, 2 and the commercial strain and on both media. The results obtained showed that a concentration of 2% sugar, 2.5 g/l yeast extract, pH 4.5 and a temperature between 25 and 35°C are the optimal conditions for cultivation in a bioreactor. The exponential phase of the specific growth rate of a strain on both media showed that it is about 0.3625 h-1 for the production of a medium based on date syrup and 0.3521 h-1 on beet molasses with a generation time equal to 1.912 h and on the medium based on date syrup, yeast consumes preferentially the reducing sugars. For the production of protein, we showed that this latter presents an exponential phase when the medium starts to run out of reducing sugars. For strain 2, the specific growth rate is about 0.261h-1 for the production on a medium based on date syrup and 0207 h-1 on beet molasses and the base medium syrup date of the yeast consumes preferentially reducing sugars. For the invertase and other metabolits, these increases rapidly after exhaustion of reducing sugars. The comparison of productivity between the three strains on the medium based on date syrup showed that the maximum value is obtained with the second strain: p = 1072 g/l/h as it is about of 0923 g/l/h for strain 1 and 0644 g/l/h for the commercial strain. Thus, isolates of date syrup are more competitive than the commercial strain and can give the same performance in a shorter time with energy gain.Keywords: date palm, fermentation, molasses, Saccharomyces, syrup
Procedia PDF Downloads 3151393 Isolation, Selection and Identification of Bacteria for Bioaugmentation of Paper Mills White Water
Authors: Nada Verdel, Tomaz Rijavec, Albin Pintar, Ales Lapanje
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Objectives: White water circuits of woodfree paper mills contain suspended, dissolved, and colloidal particles, such as cellulose, starch, paper sizings, and dyes. By closing the white water circuits, these particles start to accumulate and affect the production. Due to high amount of organic matter that scavenge radicals and adsorbs onto catalyst surfaces, treatment of white water with photocatalysis is inappropriate. The most suitable approach should be bioaugmentation-assisted bioremediation. Accordingly, objectives were: - to isolate bacteria capable of degrading organic compounds used for the papermaking process - to select the most active bacteria for bioaugmentation. Status: The state-of-the-art of bioaugmentation of pulp and paper mill effluents is mostly based on biodegradation of lignin. Whereas in white water circuits of woodfree paper mills only papermaking compounds are present. As far as one can tell from the literature, the study on degradation activities of bacteria for all possible compounds of the papermaking process is a novelty. Methodology: The main parameters of the selected white water were systematically analyzed during a period of two months. Bacteria were isolated on selective media with particular carbon source. Organic substances used as carbon source either enter white water circuits as base paper or as recycled broke. The screening of bacterial activities for starch, cellulose, latex, polyvinyl alcohol, alkyl ketene dimers, and resin acids was followed by addition of lugol. Degraders of polycyclic aromatic dyes were selected by cometabolism tests; cometabolism is simultaneous biodegradation of two compounds, in which the degradation of the second compound depends on the presence of the first. The obtained strains were identified by 16S rRNA sequencing. Findings: 335 autochthonous strains were isolated on plates with selected carbon source. The isolated strains were selected according to degradation of the particular carbon source. The ultimate degraders of cationic starch, cellulose, and sizings are Pseudomonas sp. NV-CE12-CF and Aeromonas sp. NV-RES19-BTP. The most active strains capable of degrading azo dyes are Aeromonas sp. NV-RES19-BTP and Sphingomonas sp. NV-B14-CF. Klebsiella sp. NV-Y14A-BTP degrade polycyclic aromatic direct blue 15 and also yellow dye, Agromyces sp. NV-RED15A-BF and Cellulosimicrobium sp. NV-A4-BF are specialists for whitener and Aeromonas sp. NV-RES19-BTP is general degrader of all compounds. To the white water adapted bacteria were isolated and selected according to their degradation activities for particular organic substances. Mostly isolated bacteria are specialized to lower the competition in the microbial community. Degraders of readily-biodegradable compounds do not degrade recalcitrant polycyclic aromatic dyes and vice versa. General degraders are rare.Keywords: bioaugmentation, biodegradation of azo dyes, cometabolism, smart wastewater treatment technologies
Procedia PDF Downloads 1961392 Characterization of Biosurfactants Produced by Bacteria Degrading Gasoline
Authors: Ikram Kamal, Mohamed Blaghen
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Biosurfactants are amphiphilic biological compounds consisting of hydrophobic and hydrophilic domains produced extracellularly or as part of the cell membrane by a variety of yeast, bacteria and filamentous fungi. 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). The use of biosurfactants has also been proposed for various industrial applications, such as in food additives, cosmetics, detergent formulations and in combinations with enzymes for wastewater treatment. In this study, we have investigated the potential of bacterial strains: Mannheimia haemolytica, Burkholderia cepacia and Serratia ficaria were 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: biosurfactants, Mannheimia haemolytica, biodegradability, Burkholderia cepacia, Serratia ficaria
Procedia PDF Downloads 2531391 Physiochemical and Antibacterial Assessment of Iranian Propolis Gathering in Qazvin Province
Authors: Nematollah Gheibi, Nader Divan Khosroshahi, Mahdi Mohammadi Ghanbarlou
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Introduction: Nowadays, the phenomenon of bacterial resistance is one of the most important challenge of the health community in the world. Propolis is most important production of bee colonies that collected from of various plants. So far, a lot of investigations carried out about its antibacterial effects. Material and methods: Thirty gram of propolis prepared as ethanolic extract and after different process of purification, 7.5 gr of its pure form were obtained. Propolis compounds identification was performed by TLC and VLC methods. The HPLC spectrum obtaining from propolis ethanolic extract was compared with some purified standard phenolic and flavonoid substances. Antibacterial effects of ethanol extract of purified propolis were evaluated on two strains of Staphylococcus aureus and Pseudomonas aeruginosa and their MIC was determined by the microdillution assay. Results: Ethanolic propolis extraction analyzed by TLC were resulted to confirm several phenolic and flavonoid compounds in this extract and some of the confirmed by HPLC technique. Minimum inhibitory concentration (MIC) for standard Staphylococcus aureus (ATCC25923) and Pseudomonas aeruginosa (ATCC27853) strains were obtained 2.5 mg/ml and 50 mg/ml respectively. Conclusion: Bee Propolis is a mix organic compound that has a lot of beneficial effects such as anti-bacterial that emphasized in this investigation. It is proposed as a rich source of natural phenolic and flavonoids compounds in designing of new biological resources for hygienic and medical applications.Keywords: propolis, Staphylococcus aureus, Pseudomonas aeruginosa, antibacterial
Procedia PDF Downloads 2991390 Characteristics of Acute Bacterial Prostatitis in Elderly Patients Attended in the Emergency Department
Authors: Carles Ferré, Ferran Llopis, Javier Jacob, Jordi Giol, Xavier Palom, Ignasi Bardés
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Objective: To analyze the characteristics of acute bacterial prostatitis (ABP) in elderly patients attended in the emergency department (ED). Methods: Observational and cohort study with prospective follow-up including patients with ABP presenting to the ED from January-December 2012. Data were collected for demographic variables, comorbidities, clinical and microbiological findings, treatment, outcome, and reconsultation at 30 days follow up. Findings were compared between patients ≥ 75 years (study group) and < 75 years (control group). Results: During the study period 241 episodes of ABP were included for analysis. Mean age was 62,9 ± 16 years, and 64 (26.5%) were ≥ 75 years old. A history of prostate adenoma was reported in 54 cases (22,4%), diabetes mellitus in 47 patients (19,5%) and prior manipulation of the lower urinary tract in 40 (17%). Mean symptoms duration was 3.38 ± 4.04 days, voiding symptoms were present in 176 cases (73%) and fever in 154 (64%). From 216 urine cultures, 128 were positive (59%) and 24 (17,6%) out of 136 blood cultures. Escherichia coli was the main pathogen in 58.6% of urine cultures and 64% of blood cultures (with resistant strains to fluoroquinolones in 27,7%, cotrimoxazole in 22,9% and amoxicillin/clavulanic in 27.7% of cases). Seventy patients (29%) were admitted to the hospital, and 3 died. At 30-day follow-up, 29 patients (12%) returned to the ED. In the bivariate analysis previous manipulation of the urinary tract, history of cancer, previous antibiotic treatment, resistant E. coli strains to amoxicillin-clavulanate and ciprofloxacin and extended spectrum beta-lactamase (ESBL) producers, renal impairment, and admission to the hospital were significantly more frequent (p < 0.05) among patients ≥ 75 years compared to those younger than 75 years. Conclusions: Ciprofloxacin and amoxicillin-clavulanate appear not to be good options for the empiric treatment of ABP for patients ≥ 75 years given the drug-resistance pattern in our series, and the proportion of ESBL-producing strains of E. coli should be taken into account. Awaiting bacteria identification and antibiogram from urine and/or blood cultures, treatment on an inpatient basis should be considered in older patients with ABP.Keywords: acute bacterial prostatitits, antibiotic resistance, elderly patients, emergency
Procedia PDF Downloads 3731389 Microbial Deterioration of Some Different Archaeological Objects Made from Cellulose by Bacillus Group
Authors: Mohammad Abdel Fattah Mohammad Kewisha
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Microbial deterioration of ancient materials became one of the biggest problems facing the workers in the field of cultural heritage protection because the microbial deterioration of artifacts causes detrimental effects on the aesthetic value of the monuments due to colonization, whether they are made of inorganic materials such as stone or organic like wood, textiles, wall paintings, and paper. So, the early identification of the bacterial strains that caused deterioration is the most important point for the protection of monument objects. The present study focuses on the Bacillus spp. group, which was isolated from some biodeterioration monuments from different areas of Egypt. The investigated objects in this study were made from organic materials (cellulose), paper, textile, and wood. Isolated strains were identified up to the species level biochemically. Eleven bacterial isolates were obtained from collected samples. They were taken from different archaeological objects, four microbicides, cetrimonium bromide, sodium azide, tetraethyl ammonium bromide, and dichloroxylenol, at various concentrations ranging from 25 ppm to 500 ppm. They were screened for their antibacterial activity against the Bacillus spp. isolates, and detection of Minimum inhibitory concentration (MIC). It was also necessary to indicate the ideal Minimum inhibitory concentration for each strain for the purpose of biotreatment of the infected monuments with less damaging effect on monument materials.Keywords: microbial deterioration, ancient materials, heritage protection, protection of monuments, biodeteriorative monuments
Procedia PDF Downloads 511388 Image Multi-Feature Analysis by Principal Component Analysis for Visual Surface Roughness Measurement
Authors: Wei Zhang, Yan He, Yan Wang, Yufeng Li, Chuanpeng Hao
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Surface roughness is an important index for evaluating surface quality, needs to be accurately measured to ensure the performance of the workpiece. The roughness measurement based on machine vision involves various image features, some of which are redundant. These redundant features affect the accuracy and speed of the visual approach. Previous research used correlation analysis methods to select the appropriate features. However, this feature analysis is independent and cannot fully utilize the information of data. Besides, blindly reducing features lose a lot of useful information, resulting in unreliable results. Therefore, the focus of this paper is on providing a redundant feature removal approach for visual roughness measurement. In this paper, the statistical methods and gray-level co-occurrence matrix(GLCM) are employed to extract the texture features of machined images effectively. Then, the principal component analysis(PCA) is used to fuse all extracted features into a new one, which reduces the feature dimension and maintains the integrity of the original information. Finally, the relationship between new features and roughness is established by the support vector machine(SVM). The experimental results show that the approach can effectively solve multi-feature information redundancy of machined surface images and provides a new idea for the visual evaluation of surface roughness.Keywords: feature analysis, machine vision, PCA, surface roughness, SVM
Procedia PDF Downloads 2081387 Synthesis, Antibacterial Activities, and Synergistic Effects of Novel Juglone and Naphthazarin Derivatives Against Clinical Methicillin-Resistant Staphylococcus aureus Strains
Authors: Zohra Benfodda, Valentin Duvauchelle, Chaimae Majdi, David Bénimélis, Catherine Dunyach-Remy, Patrick Meffre
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New antibiotics are necessary to treat microbial pathogens, especially ESKAPE pathogens that are becoming increasingly resistant to available treatment. Despite the medical need, the number of newly approved drugs continues to decline. The majority of antibiotics under clinical development are natural products or derivatives thereof. 43 juglone/naphthazarin derivatives were synthesized using Minisci-type direct C–H alkylation and evaluated for their antibacterial properties against various clinical and reference Gram-positive MSSA, clinical Gram-positive MRSA. Different compounds of the synthesized series showed promising activity against clinical and reference MSSA (MIC: 1–8 μg/ml) and good efficacy against clinical MRSA (MIC: 2–8 μg/ml) strains. The synergistic effects of active compounds were evaluated with reference antibiotics (vancomycin and cloxacillin), and it was found that the antibiotic combination with those active compounds efficiently enhanced the antimicrobial activity and consequently the MIC values of reference antibiotics were lowered up to 1/16th of the original MIC. These synthesized compounds did not present hemolytic activity on sheep red blood cells. In addition to the in silico prediction of ADME profile parameter which is promising and encouraging for further development.Keywords: juglone, naphthazarin, antibacterial, clinical MRSA, synergistic studies, MIC determination
Procedia PDF Downloads 1171386 Preliminary Characterization of Hericium Species Sampled in Tuscany, Italy
Authors: V. Cesaroni, C. Girometta, A. Bernicchia, M. Brusoni, F. Corana, R. M. Baiguera, C. M. Cusaro, M. L. Guglielminetti, B. Mannucci, H. Kawagishi, C. Perini, A. M. Picco, P. Rossi, E. Salerni, E. Savino
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Fungi of the genus Hericium contain various compounds with antibacterial activity, cytotoxic effect on cancer cells and bioactive molecules. Some of the active metabolites stimulate the synthesis of the Nerve Growth Factor (NGF). Recently, the effect of dietary supplement based on Hericium erinaceus on recognition memory and on hippocampal mossy fiber-CA3 neurotransmission was published. The aim of this study was to investigate the presence of Hericium species on Italian territory in order to isolate the strains for further studies and applications. The first step was to collect Hericium sporophores in Tuscany: H. alpestre Pers., H. coralloides (Scop.) Pers. and H. erinaceus (Bull.) Pers. were the species present. The strains of H. alpestre (H.a.1), H. coralloides (H.c.1) and H. erinaceus (H.e.1 & H.e.2) have been isolated in pure culture and preserved in the collection of the University of Pavia (MicUNIPV). The DNA sequences obtained from the strains were compared to other sequences found in international databases. Therefore, it was possible to construct a phylogenetic tree that highlights the clear separation in clades of the sequences and the molecular identification of our strains with the species of Hericium considered. The second step was to cultivate indoor and outdoor H. erinaceus in order to obtain as many sporophores as possible for further chemical analysis. All the procedures for H. erinaceus cultivation have been followed. Among the available recipes for indoor H. erinaceus cultivation, it was used a substrate formulation contained 70% oak sawdust, 20% rice bran, 10% wheat straw, 1% CaCO3 and 1% sucrose. The bioactive compounds present in the mycelia and in the sporophores of H. erinaceus were chemically analyzed in collaboration with the Centro Grandi Strumenti of the University of Pavia using high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS). The materials to be analyzed were previously freeze-dried and then extracted with an alcoholic procedure. Preliminary chromatographic analysis revealed the presence of potentially bioactive and structurally different secondary metabolites such as polysaccharides, erinacins, ericenones, steroids and other terpenoids. Ericenones C and D (in sporophores) and erinacin A (in mycelium) have been identified by comparison with the respective standards. These molecules are known to have effects on the Central Nervous System (CNS) cells, which is the main objective of our studies. Thanks to the high sensitivity in the detection of bioactive compounds of H. erinaceus, it will be possible to use the To obtain lyophilized mycelium and the respective culture broth, 4 small pieces (about 5 mm2) of the respective H.e.1 or H.c.1 strains, taken from the margin of growing cultures (MEA), were inoculated into 1 liter of 2% ME (malt extract, Biokar Diagnostics). The static liquid cultures were kept at 24 °C in the dark chamber and fungi grew for one month. 10 replicates for each strain have been done. The method proposed as an analytical screening protocol to determine the optimal growth conditions of the fungus and to improve the production chain of H. erinaceus. These results encourage to carry out chemical analyzes also on H. alpestre and H. coralloides in order to evaluate the presence of bioactive compounds in these two species.Keywords: Hericium species, Hercium erinaceus bioactive compounds, medicinal mushrooms, mushroom cultivation
Procedia PDF Downloads 1391385 Stenotrophomonas maltophilia: The Major Carbapenem Resistance Bacteria from Waste Water Treatment Plant of Pig Farm
Authors: Young-Ji Kim, Jin-Hyeong Park, Hong-Seok Kim, Jung-Whan Chon, Kwang-Yeop Kim, Dong-Hyeon Kim, Il-Byeong Kang, Da-Na Jeong, Jin-Hyeok Yim, Ho-Seok Jang, Kwang-Young Song, Kun-Ho Seo
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Stenotrophomonas maltophilia is one of the emerging opportunistic pathogens, and also known to have extensive drug resistance intrinsically including carbepenems which is last resort for most serious infections. One possible way for S. maltophilia to infect human is via wastewater treatment plant (WWTP). In the period between October 2016 and February 2017, effluent samples of WWTP from 3 different pig farms were collected once a month and screened for isolation of S. maltophilia. Total 16 strains of S. maltophilia were isolated and, the antibiotic susceptibility phenotypes were determined by Vitek 2 system for 16 antibiotics, ampicillin (AMP), amoxicillin/clavulanic acid (AMC), piperacillin/tazobactam (TZP), cefazolin (CZ), cefoxitin (FOX), cefotaxime (CTX), ceftazidime (CAZ), cefepime (FEP), aztreonam (AZT), ertapenem (ETP), imipenem (IMP), amikacin (AK), gentamicin (GN), ciprofloxacin (CIP), tigecycline (TGC) and trimethoprim/sulfamethoxazole (SXT). All isolates showed high resistance to AMP (100%), CZ (100%), FOX (100%), CTX (100%), CAZ (100%), FEP (94%), AZT (100%), ETP (100%), IMP (100%), AK (100%), GN (100%) whereas were susceptible to CIP (0%), TGC (0%), SXT (6%). All strains harbored at least one of the antibiotic resistance determinant such as spgM, rmlA, and rpfF. Some isolates had similar MLST (multilocus sequence typing) types with clinical isolates, suggesting WWTP could have potential role in the transmission of S. maltophilia to aquatic environment and, possibly, to humans.Keywords: Stenotrophomonas maltophilia, Carbapenem resistance, waste water treatment plant, pig farm
Procedia PDF Downloads 4551384 Improving the Biocontrol of the Argentine Stem Weevil; Using the Parasitic Wasp Microctonus hyperodae
Authors: John G. Skelly, Peter K. Dearden, Thomas W. R. Harrop, Sarah N. Inwood, Joseph Guhlin
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The Argentine stem weevil (ASW; L. bonariensis) is an economically important pasture pest in New Zealand, which causes about $200 million of damage per annum. Microctonus hyperodae (Mh), a parasite of the ASW in its natural range in South America, was introduced into New Zealand to curb the pasture damage caused by the ASW. Mh is an endoparasitic wasp that lays its eggs in the ASW halting its reproduction. Mh was initially successful at preventing ASW proliferation and reducing pasture damage. The effectiveness of Mh has since declined due to decreased parasitism rates and has resulted in increased pasture damage. Although the mechanism through which ASW has developed resistance to Mh has not been discovered, it has been proposed to be due to the different reproductive modes used by Mh and the ASW in New Zealand. The ASW reproduces sexually, whereas Mh reproduces asexually, which has been hypothesised to have allowed the ASW to ‘out evolve’ Mh. Other species within the Microctonus genus reproduce both sexually and asexually. Strains of Microctonus aethiopoides (Ma), a species closely related to Mh, reproduce either by sexual or asexual reproduction. Comparing the genomes of sexual and asexual Microctonus may allow for the identification of the mechanism of asexual reproduction and other characteristics that may improve Mh as a biocontrol agent. The genomes of Mh and three strains of Ma, two of which reproduce sexually and one reproduces asexually, have been sequenced and annotated. The French (MaFR) and Moroccan (MaMO) reproduce sexually, whereas the Irish strain (MaIR) reproduces asexually. Like Mh, The Ma strains are also used as biocontrol agents, but for different weevil species. The genomes of Mh and MaIR were subsequently upgraded using Hi-C, resulting in a set of high quality, highly contiguous genomes. A subset of the genes involved in mitosis and meiosis, which have been identified though the use of Hidden Markov Models generated from genes involved in these processes in other Hymenoptera, have been catalogued in Mh and the strains of Ma. Meiosis and mitosis genes were broadly conserved in both sexual and asexual Microctonus species. This implies that either the asexual species have retained a subset of the molecular components required for sexual reproduction or that the molecular mechanisms of mitosis and meiosis are different or differently regulated in Microctonus to other insect species in which these mechanisms are more broadly characterised. Bioinformatic analysis of the chemoreceptor compliment in Microctonus has revealed some variation in the number of olfactory receptors, which may be related to host preference. Phylogenetic analysis of olfactory receptors highlights variation, which may be able to explain different host range preferences in the Microctonus. Hi-C clustering implies that Mh has 12 chromosomes, and MaIR has 8. Hence there may be variation in gene regulation between species. Genome alignment of Mh and MaIR implies that there may be large scale genome structural variation. Greater insight into the genetics of these agriculturally important group of parasitic wasps may be beneficial in restoring or maintaining their biocontrol efficacy.Keywords: argentine stem weevil, asexual, genomics, Microctonus hyperodae
Procedia PDF Downloads 1471383 In vivo Evaluation of LAB Probiotic Potential with the Zebrafish Animal Model
Authors: Iñaki Iturria, Pasquale Russo, Montserrat Nacher-Vázquez, Giuseppe Spano, Paloma López, Miguel Angel Pardo
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Introduction: It is known that some Lactic Acid Bacteria (LAB) present an interesting probiotic effect. Probiotic bacteria stimulate host resistance to microbial pathogens and thereby aid in immune response, and modulate the host's immune responses to antigens with a potential to down-regulate hypersensitivity reactions. Therefore, probiotic therapy is valuable against intestinal infections and may be beneficial in the treatment of Inflammatory Bowel Disease (IBD). Several in vitro tests are available to evaluate the probiotic potential of a LAB strain. However, an in vivo model is required to understand the interaction between the host immune system and the bacteria. During the last few years, zebrafish (Danio rerio) has gained interest as a promising vertebrate model in this field. This organism has been extensively used to study the interaction between the host and the microbiota, as well as the host immune response under several microbial infections. In this work, we report on the use of the zebrafish model to investigate in vivo the colonizing ability and the immunomodulatory effect of probiotic LAB. Methods: Lactobacillus strains belonging to different LAB species were fluorescently tagged and used to colonize germ-free zebrafish larvae gastrointestinal tract (GIT). Some of the strains had a well-documented probiotic effect (L. acidophilus LA5); while others presented an exopolysaccharide (EPS) producing phenotype, thus allowing evaluating the influence of EPS in the colonization and immunomodulatory effect. Bacteria colonization was monitored for 72 h by direct observation in real time using fluorescent microscopy. CFU count per larva was also evaluated at different times. The immunomodulatory effect was assessed analysing the differential expression of several innate immune system genes (MyD88, NF-κB, Tlr4, Il1β and Il10) by qRT- PCR. The anti-inflammatory effect was evaluated using a chemical enterocolitis zebrafish model. The protective effect against a pathogen was also studied. To that end, a challenge test was developed using a fluorescently tagged pathogen (Vibrio anguillarum-GFP+). The progression of the infection was monitored up to 3 days using a fluorescent stereomicroscope. Mortality rates and CFU counts were also registered. Results and conclusions: Larvae exposed to EPS-producing bacteria showed a higher fluorescence and CFU count than those colonized with no-EPS phenotype LAB. In the same way, qRT-PCR results revealed an immunomodulatory effect on the host after the administration of the strains with probiotic activity. A downregulation of proinflammatory cytoquines as well as other cellular mediators of inflammation was observed. The anti-inflammatory effect was found to be particularly marked following exposure to LA% strain, as well as EPS producing strains. Furthermore, the challenge test revealed a protective effect of probiotic administration. As a matter of fact, larvae fed with probiotics showed a decrease in the mortality rate ranging from 20 to 35%. Discussion: In this work, we developed a promising model, based on the use of gnotobiotic zebrafish coupled with a bacterial fluorescent tagging in order to evaluate the probiotic potential of different LAB strains. We have successfully used this system to monitor in real time the colonization and persistence of exogenous LAB within the gut of zebrafish larvae, to evaluate their immunomodulatory effect and for in vivo competition assays. This approach could bring further insights into the complex microbial-host interactions at intestinal level.Keywords: gnotobiotic, immune system, lactic acid bacteria, probiotics, zebrafish
Procedia PDF Downloads 3231382 Extremophilic Amylases of Mycelial Fungi Strains Isolated in South Caucasus for Starch Processing
Authors: T. Urushadze, R. Khvedelidze, L. Kutateladze, M. Jobava, T. Burduli, T. Alexidze
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There is an increasing interest in reliable, wasteless, ecologically friendly technologies. About 40% of enzymes produced all over the world are used for production of syrups with high concentration of glucose-fructose. One of such technologies complies obtaining fermentable sugar glucose from raw materials containing starch by means of amylases. In modern alcohol-producing factories this process is running in two steps, involving two enzymes of different origin: bacterial α-amylase and fungal glucoamylase, as generally fungal amylases are less thermostable as compared to bacterial amylases. Selection of stable and operable at 700С and higher temperatures enzyme preparation with both α- and glucoamylase activities will allow conducting this process in one step. S. Durmishidze Institute of Biochemistry and Biotechnology owns unique collection of mycelial fungi, isolated from different ecological niches of Caucasus. As a result of screening our collection 39 strains poducing amylases were revealed. Most of them belong to the genus Aspergillus. Optimum temperatures of action of selected amylases from three producers were estableshed to be within the range 67-80°C. A. niger B-6 showed higher α-amylase activity at 67°C, and glucoamylase activity at 62°C, A. niger 6-12 showed higher α-amylase activity at 72°C, and glucoamylase activity at 65°C, Aspergillus niger p8-3 showed higher activities at 82°C and 70°C, for α-amylase and glucoamylase activities, respectively. Exhaustive hydrolysis process of starch solutions of different concentrations (3, 5, 15, and 30 %) with cultural liquid and technical preparation of Aspergillus niger p8-3 enzyme was studied. In case of low concentrations exhaustive hydrolysis of starch lasts 40–60 minutes, in case of high concentrations hydrolysis takes longer time. 98, 6% yield of glucose can be reached at incubation during 12 hours with enzyme cultural liquid and 8 hours incubation with technical preparation of the enzyme at gradual increase of temperature from 50°C to 82°C during the first 20 minutes and further decrease of temperature to 70°C. Temperature setting for high yield of glucose and high hydrolysis (pasteurizing), optimal for activity of these strains is the prerequisite to be able to carry out hydrolysis of starch to glucose in one step, and consequently, using one strain, what will be economically justified.Keywords: amylase, glucose hydrolisis, stability, starch
Procedia PDF Downloads 3441381 Isolation, Identification and Antimicrobial Susceptibility of Mycobacterium tuberculosis among Pulmonary Tuberculosis Patients
Authors: Naima Nur, Safa Islam, Saeema Islam, Faridul Alam
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Background: Drug-resistant pulmonary tuberculosis (DR-PTB), particularly multidrug-resistant tuberculosis (MDR-TB) and pre-extensive drug-resistant (pre-XDR), is a major challenge in effectively controlling TB, especially in developing. This study aimed to identify the strains of M. tuberculosis complex (MTC) and drug resistance patterns among the pulmonary tuberculosis patients. Methods: The study used a cross-sectional design, and 815 patients were recruited randomly in three study periods. In the first-period, 210 treated PTB patients, who were completed their treatment, received their diagnoses using light microscopy, fluorescence microscopy and cultured on Lowenstein-Jensen (L-J) slant, and then strains were identified as MTC by biochemical tests, and then sensitivity test in National Institute of Diseases of the Chest and Hospital. In the second-period, 220 re-treated PTB patients, who were completed their treatment, received their diagnoses using culture on L-J slant, line probe assay (LPA), and GeneXpert in the same hospital. In the last-period, during treatment, 385 MDR-PTB patients received their diagnoses using culture on L-J slant and LPA in the same hospital. Results: Among sixty-two (29.5%) PTB patients, 13% were sensitive to all first-line anti-TB drugs, 26% were MDR-TB patients, and 14.2% were pre-XDR-TB among 14 MDR-TB patients. After three years, 31% were MDR-TB among 220 re-treated PTB patients. After five years, 16.4% was pre-XDR-TB among 385 MDR-TB patients. Compared to females, male patients were significantly higher at all times. Conclusion: The current study demonstrated that in three study periods, the proportions of DR-TB, MDR-TB, and pre-XDR patients were an alarming issue and increasing daily.Keywords: multi-drug resistant, drug-resistant, pre-extensive drug resistant, pulmonary tuberculosis
Procedia PDF Downloads 491380 Factors for Entry Timing Choices Using Principal Axis Factorial Analysis and Logistic Regression Model
Authors: C. M. Mat Isa, H. Mohd Saman, S. R. Mohd Nasir, A. Jaapar
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International market expansion involves a strategic process of market entry decision through which a firm expands its operation from domestic to the international domain. Hence, entry timing choices require the needs to balance the early entry risks and the problems in losing opportunities as a result of late entry into a new market. Questionnaire surveys administered to 115 Malaysian construction firms operating in 51 countries worldwide have resulted in 39.1 percent response rate. Factor analysis was used to determine the most significant factors affecting entry timing choices of the firms to penetrate the international market. A logistic regression analysis used to examine the firms’ entry timing choices, indicates that the model has correctly classified 89.5 per cent of cases as late movers. The findings reveal that the most significant factor influencing the construction firms’ choices as late movers was the firm factor related to the firm’s international experience, resources, competencies and financing capacity. The study also offers valuable information to construction firms with intention to internationalize their businesses.Keywords: factors, early movers, entry timing choices, late movers, logistic regression model, principal axis factorial analysis, Malaysian construction firms
Procedia PDF Downloads 3711379 Discrete Element Modeling of the Effect of Particle Shape on Creep Behavior of Rockfills
Authors: Yunjia Wang, Zhihong Zhao, Erxiang Song
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Rockfills are widely used in civil engineering, such as dams, railways, and airport foundations in mountain areas. A significant long-term post-construction settlement may affect the serviceability or even the safety of rockfill infrastructures. The creep behavior of rockfills is influenced by a number of factors, such as particle size, strength and shape, water condition and stress level. However, the effect of particle shape on rockfill creep still remains poorly understood, which deserves a careful investigation. Particle-based discrete element method (DEM) was used to simulate the creep behavior of rockfills under different boundary conditions. Both angular and rounded particles were considered in this numerical study, in order to investigate the influence of particle shape. The preliminary results showed that angular particles experience more breakages and larger creep strains under one-dimensional compression than rounded particles. On the contrary, larger creep strains were observed in he rounded specimens in the direct shear test. The mechanism responsible for this difference is that the possibility of the existence of key particle in rounded particles is higher than that in angular particles. The above simulations demonstrate that the influence of particle shape on the creep behavior of rockfills can be simulated by DEM properly. The method of DEM simulation may facilitate our understanding of deformation properties of rockfill materials.Keywords: rockfills, creep behavior, particle crushing, discrete element method, boundary conditions
Procedia PDF Downloads 3101378 Dietary Pattern and Risk of Breast Cancer Among Women:a Case Control Study
Authors: Huma Naqeeb
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Epidemiological studies have shown the robust link between breast cancer and dietary pattern. There has been no previous study conducted in Pakistan, which specifically focuses on dietary patterns among breast cancer women. This study aims to examine the association of breast cancer with dietary patterns among Pakistani women. This case-control research was carried in multiple tertiary care facilities. Newly diagnosed primary breast cancer patients were recruited as cases (n = 408); age matched controls (n = 408) were randomly selected from the general population. Data on required parameters were systematically collected using subjective and objective tools. Factor and Principal Component Analysis (PCA) techniques were used to extract women’s dietary patterns. Four dietary patterns were identified based on eigenvalue >1; (i) veg-ovo-fish, (ii) meat-fat-sweet, (iii) mix (milk and its products, and gourds vegetables) and (iv) lentils - spices. Results of the multiple regressions were displayed as adjusted odds ratio (Adj. OR) and their respective confidence intervals (95% CI). After adjusted for potential confounders, veg-ovo-fish dietary pattern was found to be robustly associated with a lower risk of breast cancer among women (Adj. OR: 0.68, 95%CI: (0.46-0.99, p<0.01). The study findings concluded that attachment to the diets majorly composed of fresh vegetables, and high quality protein sources may contribute in lowering the risk of breast cancer among women.Keywords: breast cancer, dietary pattern, women, principal component analysis
Procedia PDF Downloads 1181377 Effect of Lactic Acid Bacteria Inoculant on Fermentation Quality of Sweet Sorghum Silage
Authors: Azizza Mala, Babo Fadlalla, Elnour Mohamed, Siran Wang, Junfeng Li, Tao Shao
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Sweet sorghum is considered one of the best plants for silage production and is now a more important feed crop in many countries worldwide. It is simple to ensile because of its high water-soluble carbohydrates (WSC) concentration and low buffer capacity. This study investigated the effect of adding Pediococcus acidilactici AZZ5 and Lactobacillus plantarum AZZ4 isolated from elephant grass on the fermentation quality of sweet sorghum silage. One commercial bacteria Lactobacillus Plantarum, Ecosyl MTD/1(C.B.)), and two strains were used as additives Pediococcus acidilactici (AZZ5), Lactobacillus plantarum subsp. Plantarum (AZZ4) at 6 log colony forming units (cfu)/g of fresh sweet sorghum grass in laboratory silos (1000g). After 15, 30, and 60 days, the silos for each treatment were opened. All of the isolated strains enhanced the silage quality of sweet sorghum silage compared to the control, as evidenced by significantly (P < 0.05) lower ammonia nitrogen (NH3-N) content and undesirable microbial counts, as well as greater lactic acid (L.A.) contents and lactic acid/acetic acid (LA/AA) ratios. In addition, AZZ4 performed better than all other inoculants during ensiling, as evidenced by a significant (P < 0.05) reduction in pH and ammonia-N contents and a significant increase in lactic acid contents.Keywords: fermentation, lactobacillus plantarum, lactic acid bacteria, pediococcus acidilactic, sweet sorghum
Procedia PDF Downloads 741376 Analysis of the Internal Mechanical Conditions in the Lower Limb Due to External Loads
Authors: Kent Salomonsson, Xuefang Zhao, Sara Kallin
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Human soft tissue is loaded and deformed by any activity, an effect known as a stress-strain relationship, and is often described by a load and tissue elongation curve. Several advances have been made in the fields of biology and mechanics of soft human tissue. However, there is limited information available on in vivo tissue mechanical characteristics and behavior. Confident mechanical properties of human soft tissue cannot be extrapolated from e.g. animal testing. Thus, there is need for non invasive methods to analyze mechanical characteristics of soft human tissue. In the present study, the internal mechanical conditions of the lower limb, which is subject to an external load, is studied by use of the finite element method. A detailed finite element model of the lower limb is made possible by use of MRI scans. Skin, fat, bones, fascia and muscles are represented separately and the material properties for them are obtained from literature. Previous studies have been shown to address macroscopic deformation features, e.g. indentation depth, to a large extent. However, the detail in which the internal anatomical features have been modeled does not reveal the critical internal strains that may induce hypoxia and/or eventual tissue damage. The results of the present study reveals that lumped material models, i.e. averaging of the material properties for the different constituents, does not capture regions of critical strains in contrast to more detailed models.Keywords: FEM, tissue, indentation, properties
Procedia PDF Downloads 3551375 A Quantitative Survey Research on the Development and Assessment of Attitude toward Mathematics Instrument
Authors: Soofia Malik
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The purpose of this study is to develop an instrument to measure undergraduate students’ attitudes toward mathematics (MAT) and to assess the data collected from the instrument for validity and reliability. The instrument is developed using five subscales: anxiety, enjoyment, self-confidence, value, and technology. The technology dimension is added as the fifth subscale of attitude toward mathematics because of the recent trend of incorporating online homework in mathematics courses as well as due to heavy reliance of higher education on using online learning management systems, such as Blackboard and Moodle. The sample consists of 163 (M = 82, F = 81) undergraduates enrolled in College Algebra course in the summer 2017 semester at a university in the USA. The data is analyzed to answer the research question: if and how do undergraduate students’ attitudes toward mathematics load using Principal Components Analysis (PCA)? As a result of PCA, three subscales emerged namely: anxiety/self-confidence scale, enjoyment, and value scale. After deleting the last five items or the last two subscales from the initial MAT scale, the Cronbach’s alpha was recalculated using the scores from 20 items and was found to be α = .95. It is important to note that the reliability of the initial MAT form was α = .93. This means that employing the final MAT survey form would yield consistent results in repeated uses. The final MAT form is, therefore, more reliable as compared to the initial MAT form.Keywords: college algebra, Cronbach's alpha reliability coefficient, Principal Components Analysis, PCA, technology in mathematics
Procedia PDF Downloads 1191374 Antibacterial Activity of Melaleuca Cajuputi Oil against Resistant Strain Bacteria
Authors: R. M. Noah, N. M. Nasir, M. R. Jais, M. S. S. Wahab, M. H. Abdullah, A. S. S. Raj
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Infectious diseases are getting more difficult to treat due to the resistant strains of bacteria. Current generations of antibiotics are most likely ineffective against multi-drug resistant strains bacteria. Thus, there is an urgent need in search of natural antibiotics in particular from medicinal plants. One of the common medicinal plants, Melaleuca cajuputi, has been reported to possess antibacterial properties. The study was conducted to evaluate and justify the presence of antibacterial activity of Melaleuca cajuputi essential oil (EO) against the multi-drug resistant bacteria. Clinical isolates obtained from the teaching hospital were re-assessed to confirm the exact identity of the bacteria to be tested, namely methicillin-resistant staphylococcus aureus (MRSA), carbapenem-resistant enterobacteriaceae (CRE), and extended-spectrum beta-lactamases producer (ESBLs). A well diffusion method was done to observe the inhibition zones of the essential oil against the bacteria. Minimum inhibitory concentration (MIC) was determined using the microdilution method in 96-well flat microplate. The absorbance was measured using a microplate reader. Minimum bactericidal concentration (MBC) was performed using the agar medium method. The zones of inhibition produced by the EO against MRSA, CRE, and ESBL were comparable to that of generic antibiotics used, gentamicin and augmentin. The MIC and MBC results highlighted the antimicrobial efficacy of the EO. The outcome of this study indicated that the EO of Melaleuca cajuputi had antibacterial activity on the multi-drug resistant bacteria. This finding was eventually substantiated by electron microscopy work.Keywords: melaleuca cajuputi, antibacterial, resistant bacteria, essential oil
Procedia PDF Downloads 1171373 Atmospheric Polycyclic Aromatic Hydrocarbons (PAHs) in Rural and Urban of Central Taiwan
Authors: Shih Yu Pan, Pao Chen Hung, Chuan Yao Lin, Charles C.-K. Chou, Yu Chi Lin, Kai Hsien Chi
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This study analyzed 16 atmospheric PAHs species which were controlled by USEPA and IARC. To measure the concentration of PAHs, four rural sampling sites and two urban sampling sites were selected in Central Taiwan during spring and summer. In central Taiwan, the rural sampling stations were located in the downstream of Da-An River, Da-Jang River, Wu River and Chuo-shui River. On the other hand, the urban sampling sites were located in Taichung district and close to the roadside. Ambient air samples of both vapor phase and particle phase of PAHs compounds were collected using high volume sampling trains (Analitica). The sampling media were polyurethane foam (PUF) with XAD2 and quartz fiber filters. Diagnostic ratio, Principal component analysis (PCA), Positive Matrix Factorization (PMF) models were used to evaluate the apportionment of PAHs in the atmosphere and speculate the relative contribution of various emission sources. Because of the high temperature and low wind speed, high PAHs concentration in the atmosphere was observed. The total PAHs concentration, especially in vapor phase, had significant change during summer. During the sampling periods the total PAHs concentration of atmospheric at four rural and two urban sampling sites in spring and summer were 3.70±0.40 ng/m3,3.40±0.63 ng/m3,5.22±1.24 ng/m3,7.23±0.37 ng/m3,7.46±2.36 ng/m3,6.21±0.55 ng/m3 ; 15.0± 0.14 ng/m3,18.8±8.05 ng/m3,20.2±8.58 ng/m3,16.1±3.75 ng/m3,29.8±10.4 ng/m3,35.3±11.8 ng/m3, respectively. In order to identify PAHs sources, we used diagnostic ratio to classify the emission sources. The potential sources were diesel combustion and gasoline combustion in spring and summer, respectively. According to the principal component analysis (PCA), the PC1 and PC2 had 23.8%, 20.4% variance and 21.3%, 17.1% variance in spring and summer, respectively. Especially high molecular weight PAHs (BaP, IND, BghiP, Flu, Phe, Flt, Pyr) were dominated in spring when low molecular weight PAHs (AcPy, Ant, Acp, Flu) because of the dominating high temperatures were dominated in the summer. Analysis by using PMF model found the sources of PAHs in spring were stationary sources (34%), vehicle emissions (24%), coal combustion (23%) and petrochemical fuel gas (19%), while in summer the emission sources were petrochemical fuel gas (34%), the natural environment of volatile organic compounds (29%), coal combustion (19%) and stationary sources (18%).Keywords: PAHs, source identification, diagnostic ratio, principal component analysis, positive matrix factorization
Procedia PDF Downloads 2631372 Unifying RSV Evolutionary Dynamics and Epidemiology Through Phylodynamic Analyses
Authors: Lydia Tan, Philippe Lemey, Lieselot Houspie, Marco Viveen, Darren Martin, Frank Coenjaerts
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Introduction: Human respiratory syncytial virus (hRSV) is the leading cause of severe respiratory tract infections in infants under the age of two. Genomic substitutions and related evolutionary dynamics of hRSV are of great influence on virus transmission behavior. The evolutionary patterns formed are due to a precarious interplay between the host immune response and RSV, thereby selecting the most viable and less immunogenic strains. Studying genomic profiles can teach us which genes and consequent proteins play an important role in RSV survival and transmission dynamics. Study design: In this study, genetic diversity and evolutionary rate analysis were conducted on 36 RSV subgroup B whole genome sequences and 37 subgroup A genome sequences. Clinical RSV isolates were obtained from nasopharyngeal aspirates and swabs of children between 2 weeks and 5 years old of age. These strains, collected during epidemic seasons from 2001 to 2011 in the Netherlands and Belgium by either conventional or 454-sequencing. Sequences were analyzed for genetic diversity, recombination events, synonymous/non-synonymous substitution ratios, epistasis, and translational consequences of mutations were mapped to known 3D protein structures. We used Bayesian statistical inference to estimate the rate of RSV genome evolution and the rate of variability across the genome. Results: The A and B profiles were described in detail and compared to each other. Overall, the majority of the whole RSV genome is highly conserved among all strains. The attachment protein G was the most variable protein and its gene had, similar to the non-coding regions in RSV, more elevated (two-fold) substitution rates than other genes. In addition, the G gene has been identified as the major target for diversifying selection. Overall, less gene and protein variability was found within RSV-B compared to RSV-A and most protein variation between the subgroups was found in the F, G, SH and M2-2 proteins. For the F protein mutations and correlated amino acid changes are largely located in the F2 ligand-binding domain. The small hydrophobic phosphoprotein and nucleoprotein are the most conserved proteins. The evolutionary rates were similar in both subgroups (A: 6.47E-04, B: 7.76E-04 substitution/site/yr), but estimates of the time to the most recent common ancestor were much lower for RSV-B (B: 19, A: 46.8 yrs), indicating that there is more turnover in this subgroup. Conclusion: This study provides a detailed description of whole RSV genome mutations, the effect on translation products and the first estimate of the RSV genome evolution tempo. The immunogenic G protein seems to require high substitution rates in order to select less immunogenic strains and other conserved proteins are most likely essential to preserve RSV viability. The resulting G gene variability makes its protein a less interesting target for RSV intervention methods. The more conserved RSV F protein with less antigenic epitope shedding is, therefore, more suitable for developing therapeutic strategies or vaccines.Keywords: drug target selection, epidemiology, respiratory syncytial virus, RSV
Procedia PDF Downloads 4081371 Antibacterial Activity of Rosmarinus officinalis (Rosemary) and Murraya koenigii (Curry Leaves) against Multidrug Resistant S. aureus and Coagulase Negative Staphylococcus Species
Authors: Asma Naim, Warda Mushtaq
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Staphylococcus species are the most versatile and adaptive organism. They are widespread and naturally found on the skin, mucosa and nose in humans. Among these, Staphylococcus aureus is the most important species. These organisms act as opportunistic pathogens and can infect various organs of the host, causing minor skin infection to severe toxin mediated diseases, and life threatening nosocomial infections. Staphylococcus aureus has acquired resistance against β-lactam antibiotics by the production of β-lactamase, and Methicillin-Resistant Staphylococcus aureus (MRSA) strains have also been reported with increasing frequency. MRSA strains have been associated with nosocomial as well as community acquired infections. Medicinal plants have enormous potential as antimicrobial substances and have been used in traditional medicine. Search for medicinally valuable plants with antimicrobial activity is being emphasized due to increasing antibiotic resistance in bacteria. In the present study, the antibacterial potential of Rosmarinus officinalis (Rosemary) and Murraya koenigii (curry leaves) was evaluated. These are common household herbs used in food as enhancer of flavor and aroma. The crude aqueous infusion, decoction and ethanolic extracts of curry leaves and rosemary and essential oil of rosemary were investigated in the present study for antibacterial activity against multi-drug resistant Staphylococcus strains using well diffusion method. In the present study, 60 Multi-drug resistant clinical isolates of S. aureus (43) and Coagulase Negative Staphylococci (CoNS) (17) were screened against different concentrations of crude extracts of Rosmarinus officinalis and Murraya koenigii. Out of these 60 isolates, 43 were sensitive to the aqueous infusion of rosemary; 23 to aqueous decoction and 58 to ethanolic extract whereas, 24 isolates were sensitive to the essential oil. In the case of the curry leaves, no antibacterial activity was observed in aqueous infusion and decoction while only 14 isolates were sensitive to the ethanolic extract. The aqueous infusion of rosemary (50% concentration) exhibited a zone of inhibition of 21(±5.69) mm. against CoNS and 17(±4.77) mm. against S. aureus, the zone of inhibition of 50% concentration of aqueous decoction of rosemary was also larger against CoNS 17(±5.78) mm. then S. aureus 13(±6.91) mm. and the 50% concentrated ethanolic extract showed almost similar zone of inhibition in S. aureus 22(±3.61) mm. and CoNS 21(±7.64) mm. whereas, the essential oil of rosemary showed greater zone of inhibition against S. aureus i.e., 16(±4.67) mm. while CoNS showed 15(±6.94) mm. These results show that ethanolic extract of rosemary has significant antibacterial activity. Aqueous infusion and decoction of curry leaves revealed no significant antibacterial potential against all Staphylococcal species and ethanolic extract also showed only a weak response. Staphylococcus strains were susceptible to crude extracts and essential oil of rosemary in a dose depend manner, where the aqueous infusion showed highest zone of inhibition and ethanolic extract also demonstrated antistaphylococcal activity. These results demonstrate that rosemary possesses antistaphylococcal activity.Keywords: antibacterial activity, curry leaves, multidrug resistant, rosemary, S. aureus
Procedia PDF Downloads 2391370 Marine Phytoplankton and Zooplankton from the North-Eastern Bay of Bengal, Bangladesh
Authors: Mahmudur Rahman Khan, Saima Sharif Nilla, Kawser Ahmed, Abdul Aziz
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The marine phyto and zooplankton of the extreme north-eastern part of the Bay of Bengal, off the coast of Bangladesh have been studied. Relative occurrence of phyto and zooplankton and their relationship with physico-chemical conditions (f.e. temperature, salinity, dissolved oxygen, carbonate, phosphate, and sulphate) of the water and Shannon-Weiber diversity indices were also studied. The phytoplankton communities represented by 25 genera with 69 species of Bacillariophyceae, 5 genera with 12 species of Dinophyceae and 6 genera with 16 species of Chlorophyceae have been found. A total of 24 genera of 25 species belonging to Protozoa, Coelenterata, Chaetognatha, Nematoda, Cladocera, Copepoda, and decapoda have been recorded. In addition, the average phytoplankton was 80% of all collections, whereas the zooplankton was 20%, Z ratio of about 4:1. The total numbers of plankton individuals per liter were generally higher during low tide than those of high one. Shannon-Weiber diversity indices were highest (3.675 for phytoplankton and 3.021 for zooplankton) in the north-east part and lowest (1.516 for phytoplankton and 1.302 for zooplankton) in the south-east part of the study area. Principal Component Analysis (PCA) showed the relationship between pH and some species of phyto and zooplankton where all diatoms and copepods have showed positive correlation and dinoflagellates showed negative correlation with pH.Keywords: plankton presence, shannon-weiber diversity index, principal component analysis, Bay of Bengal
Procedia PDF Downloads 6451369 A Comparative Evaluation of Broiler Strains Chickens, Arbor Acres, and Ross in Experimental Coccidiosis
Authors: S. S. R. Shojaei, S. Kord Afshari
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The study was initiated to compare the production and defecation of Eimerial oocysts of two internationally reputed broiler strains under the local environmental and management conditions. 40 one-day old male chickens of Arbor Acres strain and ROSS strain (20 chicks from each strain) used in this study and were divided randomly into four control and challenge groups. Feed and water were provided for ad libitum consumption. At 15 d of age, chickens of challenge groups (from each strain) were individually inoculated with a mixture of 50000 of sporulated oocysts of 4 species including of E. acervulina (20%), E. maxima (40%), E. tenella (25%) and E. necatrix (15%) and also from the fourth day after Eimerial challenge, faecal droppings (litter samples) were collected 10 days consecutively for counting oocyst per gram (OPG). The results indicated that in the challenge groups, there was an increasing process of OPG in days of 4 to 7 post challenging and the pick level of OPG was seen at seventh day after challenging. From day 8 to 9, decreasing of OPG was happened. This decreasing continues with mild, fast and mild process to day of 13. Respectively and totally the average of OPG in the Arbor Acres group was lower than the group Ross in all days post inoculation and this difference was significant according to t-test. According to the obtained results in this study and since oocyst index almost always is considered as one of the most important indicators for coccidiosis evaluation, it can be realized that in the same surveillance condition the regarding the severity evaluation of coccidiosis, Arbor Acres strain broilers shed less oocysts than Ross strain broilers.Keywords: arbor acres, ross, coccidiosis, OPG
Procedia PDF Downloads 499