Search results for: Pathogenic microorganisms
Commenced in January 2007
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Edition: International
Paper Count: 953

Search results for: Pathogenic microorganisms

203 Production and Evaluation of Physicochemical, Nutritional, Sensorial and Microbiological Properties of Mixed Fruit Juice Blend Prepared from Apple, Orange and Mosambi

Authors: Himalaya Patir, Bitupon Baruah, Sanjay Gayary, Subhajit Ray

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In recent age significant importance is given for the development of nutritious and health beneficial foods. Fruit juices collected from different fruits when blended that improves not only the physicochemical and nutritional properties but also enhance the sensorial or organoleptic properties. The study was carried out to determine the physico-chemical, nutritional, microbiological analysis and sensory evaluation of mixed fruit juice blend. Juice of orange (Citrus sinensis), apple (Malus domestica), mosambi (Citrus limetta) were blended in the ratio of sample-I (30% apple:30% orange:40% mosambi), sample-II ( 40% apple :30% orange :30% mosambi), sample-III (30% apple :40% orange :30% mosambi) , sample-IV (50% apple :30% orange :20% mosambi), sample-V (30% apple:20% orange:50% mosambi), sample-VI (20% apple :50% orange :30% mosambi) to evaluate all quality characteristics. Their colour characteristics in terms of hue angle, chroma and colour difference (∆E) were evaluated. The physico-chemical parameters analysis carried out were total soluble solids (TSS), total titratable acidity (TTA), pH, acidity (FA), volatile acidity (VA), pH, and vitamin C. There were significant differences (p˂0.05) in the TSS of the samples. However, sample-V (30% apple: 20% orange: 50% mosambi) provides the highest TSS of 9.02gm and significantly differed from other samples (p˂0.05). Sample-IV (50% apple: 30% orange: 20% mosambi) was shown the highest titratable acidity (.59%) in comparison to other samples. The highest value of pH was found as 5.01 for sample-IV (50% apple: 30% orange: 20% mosambi). Sample-VI (20% apple: 50% orange :30% mosambi) blend has the highest hue angle, chroma and colour changes of 72.14,25.29 and 54.48 and vitamin C, i.e. Ascorbic acid (.33g/l) content compared to other samples. The nutritional compositions study showed that, sample- VI (20% apple: 50% orange: 30% mosambi) has the significantly higher carbohydrate (51.67%), protein (.78%) and ash (1.24%) than other samples, while sample-V (30% apple: 20% orange: 50% mosambi) has higher dietary fibre (12.84%) and fat (2.82%) content. Microbiological analysis of all samples in terms of total plate count (TPC) ranges from 44-60 in 101 dilution and 4-5 in 107 dilutions and was found satisfactory. Moreover, other pathogenic bacterial count was found nil. The general acceptability of the mixed fruit juice blend samples were moderately liked by the panellists, and sensorial quality studies showed that sample-V (30% apple: 20% orange: 50% mosambi) contains highest overall acceptability of 8.37 over other samples and can be considered good for consumption.

Keywords: microbiological, nutritional, physico-chemical, sensory properties

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202 A Program of Data Analysis on the Possible State of the Antibiotic Resistance in Bangladesh Environment in 2019

Authors: S. D. Kadir

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Background: Antibiotics have always been at the centrum of the revolution of modern microbiology. Micro-organisms and its pathogenicity, resistant organisms, inappropriate or over usage of various types of antibiotic agents are fuelled multidrug-resistant pathogenic organisms. Our present time review report mainly focuses on the therapeutic condition of antibiotic resistance and the possible roots behind the development of antibiotic resistance in Bangladesh in 2019. Methodology: The systemic review has progressed through a series of research analyses on various manuscripts published on Google Scholar, PubMed, Research Gate, and collected relevant information from established popular healthcare and diagnostic center and its subdivisions all over Bangladesh. Our research analysis on the possible assurance of antibiotic resistance been ensured by the selective medical reports and on random assay on the extent of individual antibiotic in 2019. Results: 5 research articles, 50 medical report summary, and around 5 patients have been interviewed while going through the estimation process. We have prioritized research articles where the research analysis been performed by the appropriate use of the Kirby-Bauer method. Kirby-Bauer technique is preferred as it provides greater efficiency, ensures lower performance expenditure, and supplies greater convenience and simplification in the application. In most of the reviews, clinical and laboratory standards institute guidelines were strictly followed. Most of our reports indicate significant resistance shown by the Beta-lactam drugs. Specifically by the derivatives of Penicillin's, Cephalosporin's (rare use of the first generation Cephalosporin and overuse of the second and third generation of Cephalosporin and misuse of the fourth generation of Cephalosporin), which are responsible for almost 67 percent of the bacterial resistance. Moreover, approximately 20 percent of the resistance was due to the fact of drug pumping from the bacterial cell by tetracycline and sulphonamides and their derivatives. Conclusion: 90 percent of the approximate antibiotic resistance is due to the usage of relative and true broad-spectrum antibiotics. The environment has been created by the following circumstances where; the excessive usage of broad-spectrum antibiotics had led to a condition where the disruption of native bacteria and a series of anti-microbial resistance causing a disturbance of the surrounding environments in medium, leading to a state of super-infection.

Keywords: antibiotics, antibiotic resistance, Kirby Bauer method, microbiology

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201 Morphology Analysis of Apple-Carrot Juice Treated by Manothermosonication (MTS) and High Temperature Short Time (HTST) Processes

Authors: Ozan Kahraman, Hao Feng

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Manothermosonication (MTS), which consists of the simultaneous application of heat and ultrasound under moderate pressure (100-700 kPa), is one of the technologies which destroy microorganisms and inactivates enzymes. Transmission electron microscopy (TEM) is a microscopy technique in which a beam of electrons is transmitted through an ultra-thin specimen, interacting with the specimen as it passes through it. The environmental scanning electron microscope or ESEM is a scanning electron microscope (SEM) that allows for the option of collecting electron micrographs of specimens that are "wet," uncoated. These microscopy techniques allow us to observe the processing effects on the samples. This study was conducted to investigate the effects of MTS and HTST treatments on the morphology of apple-carrot juices by using TEM and ESEM microscopy. Apple-carrot juices treated with HTST (72 0C, 15 s), MTS 50 °C (60 s, 200 kPa), and MTS 60 °C (30 s, 200 kPa) were observed in both ESEM and TEM microscopy. For TEM analysis, a drop of the solution dispersed in fixative solution was put onto a Parafilm ® sheet. The copper coated side of the TEM sample holder grid was gently laid on top of the droplet and incubated for 15 min. A drop of a 7% uranyl acetate solution was added and held for 2 min. The grid was then removed from the droplet and allowed to dry at room temperature and presented into the TEM. For ESEM analysis, a critical point drying of the filters was performed using a critical point dryer (CPD) (Samdri PVT- 3D, Tousimis Research Corp., Rockville, MD, USA). After the CPD, each filter was mounted onto a stub and coated with gold/palladium with a sputter coater (Desk II TSC Denton Vacuum, Moorestown, NJ, USA). E.Coli O157:H7 cells on the filters were observed with an ESEM (Philips XL30 ESEM-FEG, FEI Co., Eindhoven, The Netherland). ESEM (Environmental Scanning Electron Microscopy) and TEM (Transmission Electron Microscopy) images showed extensive damage for the samples treated with MTS at 50 and 60 °C such as ruptured cells and breakage on cell membranes. The damage was increasing with increasing exposure time.

Keywords: MTS, HTST, ESEM, TEM, E.COLI O157:H7

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200 Enzyme Involvement in the Biosynthesis of Selenium Nanoparticles by Geobacillus wiegelii Strain GWE1 Isolated from a Drying Oven

Authors: Daniela N. Correa-Llantén, Sebastián A. Muñoz-Ibacache, Mathilde Maire, Jenny M. Blamey

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The biosynthesis of nanoparticles by microorganisms, on the contrary to chemical synthesis, is an environmentally-friendly process which has low energy requirements. In this investigation, we used the microorganism Geobacillus wiegelii, strain GWE1, an aerobic thermophile belonging to genus Geobacillus, isolated from a drying oven. This microorganism has the ability to reduce selenite evidenced by the change of color from colorless to red in the culture. Elemental analysis and composition of the particles were verified using transmission electron microscopy and energy-dispersive X-ray analysis. The nanoparticles have a defined spherical shape and a selenium elemental state. Previous experiments showed that the presence of the whole microorganism for the reduction of selenite was not necessary. The results strongly suggested that an intracellular NADPH/NADH-dependent reductase mediates selenium nanoparticles synthesis under aerobic conditions. The enzyme was purified and identified by mass spectroscopy MALDI-TOF TOF technique. The enzyme is a 1-pyrroline-5-carboxylate dehydrogenase. Histograms of nanoparticles sizes were obtained. Size distribution ranged from 40-160 nm, where 70% of nanoparticles have less than 100 nm in size. Spectroscopic analysis showed that the nanoparticles are composed of elemental selenium. To analyse the effect of pH in size and morphology of nanoparticles, the synthesis of them was carried out at different pHs (4.0, 5.0, 6.0, 7.0, 8.0). For thermostability studies samples were incubated at different temperatures (60, 80 and 100 ºC) for 1 h and 3 h. The size of all nanoparticles was less than 100 nm at pH 4.0; over 50% of nanoparticles have less than 100 nm at pH 5.0; at pH 6.0 and 8.0 over 90% of nanoparticles have less than 100 nm in size. At neutral pH (7.0) nanoparticles reach a size around 120 nm and only 20% of them were less than 100 nm. When looking at temperature effect, nanoparticles did not show a significant difference in size when they were incubated between 0 and 3 h at 60 ºC. Meanwhile at 80 °C the nanoparticles suspension lost its homogeneity. A change in size was observed from 0 h of incubation at 80ºC, observing a size range between 40-160 nm, with 20% of them over 100 nm. Meanwhile after 3 h of incubation at size range changed to 60-180 nm with 50% of them over 100 nm. At 100 °C the nanoparticles aggregate forming nanorod structures. In conclusion, these results indicate that is possible to modulate size and shape of biologically synthesized nanoparticles by modulating pH and temperature.

Keywords: genus Geobacillus, NADPH/NADH-dependent reductase, selenium nanoparticles, biosynthesis

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199 Electrochemical Bioassay for Haptoglobin Quantification: Application in Bovine Mastitis Diagnosis

Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo

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Mastitis is the most relevant inflammatory disease in cattle, affecting the animal health and causing important economic losses on dairy farms. This disease takes place in the mammary gland or udder when some opportunistic microorganisms, such as Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis, etc., invade the teat canal. According to the severity of the inflammation, mastitis can be classified as sub-clinical, clinical and chronic. Standard methods for mastitis detection include counts of somatic cells, cell culture, electrical conductivity of the milk, and California test (evaluation of “gel-like” matrix consistency after cell lysed with detergents). However, these assays present some limitations for accurate detection of subclinical mastitis. Currently, haptoglobin, an acute phase protein, has been proposed as novel and effective biomarker for mastitis detection. In this work, an electrochemical biosensor based on polydopamine-modified magnetic nanoparticles (MNPs@pDA) for haptoglobin detection is reported. Thus, MNPs@pDA has been synthesized by our group and functionalized with hemoglobin due to its high affinity to haptoglobin protein. The protein was labeled with specific antibodies modified with alkaline phosphatase enzyme for its electrochemical detection using an electroactive substrate (1-naphthyl phosphate) by differential pulse voltammetry. After the optimization of assay parameters, the haptoglobin determination was evaluated in milk. The strategy presented in this work shows a wide range of detection, achieving a limit of detection of 43 ng/mL. The accuracy of the strategy was determined by recovery assays, being of 84 and 94.5% for two Hp levels around the cut off value. Milk real samples were tested and the prediction capacity of the electrochemical biosensor was compared with a Haptoglobin commercial ELISA kit. The performance of the assay has demonstrated this strategy is an excellent and real alternative as screen method for sub-clinical bovine mastitis detection.

Keywords: bovine mastitis, haptoglobin, electrochemistry, magnetic nanoparticles, polydopamine

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198 Nutritional Characteristics, Phytochemical and Antimicrobial Properties Vaccinium Pavifolium (Ericacea) Leaf Protein Concentrates

Authors: Sodamade A., Bolaji K. A.

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Problems associated with protein malnutrition are still prevalent in third-world countries, leading to the constant search for plants that could serve as nutrients and medicinal purposes. Huckleberry is one of the plants that has been proven useful locally in the treatment of numerous ailments and diseases. A fresh sample of the plant (Vaccinium pavifolium) was collected from a vegetable garden situated near the Erelu dam of the Emmanuel Alayande College of Education Campus, Oyo. The sample was authenticated at the Forestry Research Institute of Nigeria (FRIN) Ibadan. The leaves of the plant were plucked and processed for leaf protein concentrates before proximate composition, mineral analysis phytochemical and antimicrobial properties were determined using a standard method of analysis. The results of proximate constituents showed; moisture content; 9.89±0.051g/100g, Ash; 3.23±0.12g/100g, crude fat; 3.96±0.11g/100g and 61.27±0.56g/100g of Nitrogen free extractive. The mineral analysis of the sample showed; Mg; 0.081±0.00mg/100g, Ca; 42.30±0.05mg/100g, Na; 27.57±0.09mg/100g, K; 6.81±0.01mg/100g, P; 8.90±0.03mg/100g, Fe; 0.51±0.00mg/100g, Zn; 0.021±0.00mg/100g, Cd; 0.04±0.04mg/100g, Pb; 0.002±0.00mg/100g, Cr; 0.041±0.00mg/100g Cadmium and Mercury were not detected in the sample. The result of phytochemical analysis of leaf protein concentrates of the Huckleberry showed the presence of Alkaloid, Saponin, Flavonoid, Tanin, Coumarin, Steroids, Terpenoids, Cardiac glycosides, Glycosides, Quinones, Anthocyanin, phytosterols, and phenols. Ethanolic extracts of the Vaccinium parvifolium L. leaf protein concentrates showed that it contains bioactive compounds that are capable of combating the following microorganisms; Staphylococcus aureus, Streptococcus pyogenes, Streptococcus faecalis, Pseudomonas aeruginosa, Klebisialae pneumonia and Proteus mirabilis. The results of the analysis of Vaccinium parvifolium L. leaf protein concentrates showed that the sample contains valuable nutrient and mineral constituents, and phytochemical compounds that could make the sample useful for medicinal activities.

Keywords: leaf protein concentrates, vaccinium parvifolium, nutritional characteristics, mineral composition, antimicrobial activity

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197 Genome Sequencing and Analysis of the Spontaneous Nanosilver Resistant Bacterium Proteus mirabilis Strain scdr1

Authors: Amr Saeb, Khalid Al-Rubeaan, Mohamed Abouelhoda, Manojkumar Selvaraju, Hamsa Tayeb

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Background: P. mirabilis is a common uropathogenic bacterium that can cause major complications in patients with long-standing indwelling catheters or patients with urinary tract anomalies. In addition, P. mirabilis is a common cause of chronic osteomyelitis in diabetic foot ulcer (DFU) patients. Methodology: P. mirabilis SCDR1 was isolated from a diabetic ulcer patient. We examined P. mirabilis SCDR1 levels of resistance against nano-silver colloids, the commercial nano-silver and silver containing bandages and commonly used antibiotics. We utilized next generation sequencing techniques (NGS), bioinformatics, phylogenetic analysis and pathogenomics in the identification and characterization of the infectious pathogen. Results: P. mirabilis SCDR1 is a multi-drug resistant isolate that also showed high levels of resistance against nano-silver colloids, nano-silver chitosan composite and the commercially available nano-silver and silver bandages. The P. mirabilis-SCDR1 genome size is 3,815,621 bp with G+C content of 38.44%. P. mirabilis-SCDR1 genome contains a total of 3,533 genes, 3,414 coding DNA sequence genes, 11, 10, 18 rRNAs (5S, 16S, and 23S), and 76 tRNAs. Our isolate contains all the required pathogenicity and virulence factors to establish a successful infection. P. mirabilis SCDR1 isolate is a potential virulent pathogen that despite its original isolation site, wound, it can establish kidney infection and its associated complications. P. mirabilis SCDR1 contains several mechanisms for antibiotics and metals resistance including, biofilm formation, swarming mobility, efflux systems, and enzymatic detoxification. Conclusion: P. mirabilis SCDR1 is the spontaneous nano-silver resistant bacterial strain. P. mirabilis SCDR1 strain contains all reported pathogenic and virulence factors characteristic for the species. In addition, it possesses several mechanisms that may lead to the observed nano-silver resistance.

Keywords: Proteus mirabilis, multi-drug resistance, silver nanoparticles, resistance, next generation sequencing techniques, genome analysis, bioinformatics, phylogeny, pathogenomics, diabetic foot ulcer, xenobiotics, multidrug resistance efflux, biofilm formation, swarming mobility, resistome, glutathione S-transferase, copper/silver efflux system, altruism

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196 GC-MS-Based Untargeted Metabolomics to Study the Metabolism of Pectobacterium Strains

Authors: Magdalena Smoktunowicz, Renata Wawrzyniak, Malgorzata Waleron, Krzysztof Waleron

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Pectobacterium spp. were previously classified into the Erwinia genus founded in 1917 to unite at that time all Gram-negative, fermentative, nonsporulating and peritrichous flagellated plant pathogenic bacteria. After work of Waldee (1945), on Approved Lists of Bacterial Names and bacteriology manuals in 1980, they were described either under the species named Erwinia or Pectobacterium. The Pectobacterium genus was formally described in 1998 of 265 Pectobacterium strains. Currently, there are 21 species of Pectobacterium bacteria, including Pectobacterium betavasculorum since 2003, which caused soft rot on sugar beet tubers. Based on the biochemical experiments carried out for this, it is known that these bacteria are gram-negative, catalase-positive, oxidase-negative, facultatively anaerobic, using gelatin and causing symptoms of soft rot on potato and sugar beet tubers. The mere fact of growing on sugar beet may indicate a metabolism characteristic only for this species. Metabolomics, broadly defined as the biology of the metabolic systems, which allows to make comprehensive measurements of metabolites. Metabolomics, in combination with genomics, are complementary tools for the identification of metabolites and their reactions, and thus for the reconstruction of metabolic networks. The aim of this study was to apply the GC-MS-based untargeted metabolomics to study the metabolism of P. betavasculorum in different growing conditions. The metabolomic profiles of biomass and biomass media were determined. For sample preparation the following protocol was used: extraction with 900 µl of methanol: chloroform: water mixture (10: 3: 1, v: v) were added to 900 µl of biomass from the bottom of the tube and up to 900 µl of nutrient medium from the bacterial biomass. After centrifugation (13,000 x g, 15 min, 4oC), 300µL of the obtained supernatants were concentrated by rotary vacuum and evaporated to dryness. Afterwards, two-step derivatization procedure was performed before GC-MS analyses. The obtained results were subjected to statistical calculations with the use of both uni- and multivariate tests. The obtained results were evaluated using KEGG database, to asses which metabolic pathways are activated and which genes are responsible for it, during the metabolism of given substrates contained in the growing environment. The observed metabolic changes, combined with biochemical and physiological tests, may enable pathway discovery, regulatory inference and understanding of the homeostatic abilities of P. betavasculorum.

Keywords: GC-MS chromatograpfy, metabolomics, metabolism, pectobacterium strains, pectobacterium betavasculorum

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195 Control of the Sustainability of Fresh Cheese in Order to Extend the Shelf-Life of the Product

Authors: Radovan Čobanović, Milica Rankov Šicar

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The fresh cheese is in the group of perishable food which cannot be kept a long period of time. The study of sustainability have been done in order to extend the shelf-life of the product which was 15 days. According to the plan of sustainability it was defined that 35 samples had to be stored for 30 days at 2°C−6°C and analyzed every 7th day from the day of reception until 30th day. Shelf life of the cheese has expired during the study of sustainability in the period between 15th and 30th day of analyses. Cheese samples were subjected to sensory analysis (appearance, odor, taste, color, aroma) and bacteriological analyzes (Listeria monocytogenes, Salmonella spp., Bacillus cereus, Staphylococcus aureus and total plate count) according to Serbian state regulation. All analyses were tested according to ISO methodology: sensory analysis ISO 6658, Listeria monocytogenes ISO 11 290-1, Salmonella spp ISO 6579, Bacillus cereus ISO 7932, Staphylococcus aureus ISO 6888-1, and total plate count ISO 4833. Analyses showed that after fifteen days of storage at a temperature defined by the manufacturers and within the product's shelf life, the cheese did not have any noticeable changes in sensory characteristics. Smell and taste are unaffected there was no separation of whey and there was not presence of strange smell or taste. As far as microbiological analyses are concerned neither one pathogen was detected and total plate count was at level of 103 cfu/g. After expiry of shelf life in a period of 15th and 30th day of storage, the analysis showed that there was a separation of whey on the surface. Along the edge of the container was present a dried part of cheese and sour-milky smell and taste were very weakly expressed. Concerning the microbiological analyses there still were not positive results for pathogen microorganisms but the total plate count was at a level of 106cfu/g. Based on the obtained results it can be concluded that this product cannot have longer shelf life than shelf life which is already defined because there are a sensory changes that would certainly have influence on decision of customers when purchase of this product is concerned.

Keywords: sustainability, fresh cheese, shelf-life, product

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194 Enhanced Methane Yield from Organic Fraction of Municipal Solid Waste with Coconut Biochar as Syntrophic Metabolism Biostimulant

Authors: Maria Altamirano, Alfonso Duran

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Biostimulation has recently become important in order to improve the stability and performance of the anaerobic digestion (AD) process. This strategy involves the addition of nutrients or supplements to improve the rate of degradation of a native microbial consortium. With the aim of biostimulate sytrophism between secondary fermenting bacteria and methanogenic archaea, improving metabolite degradation and efficient conversion to methane, the addition of conductive materials, mainly carbon based have been studied. This research seeks to highlight the effect that coconut biochar (CBC) has on the metanogenic conversion of the organic fraction of municipal solid waste (OFMSW), analyzing the surface chemistry properties that give biochar its capacity to serve as a redox mediator in the anaerobic digestion process. The biochar characterization techniques were electrical conductivity (EC) scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), Fourier Transform Infrared Transmission Spectroscopy (FTIR) and Cyclic Voltammetry (CV). Effect of coconut biochar addition was studied using Authomatic Methane Potential Test System (AMPTS II) applying a one-way variance analysis to determine the dose that leads to higher methane performance. The surface chemistry of the CBC could confer properties that enhance the AD process, such as the presence of alkaline and alkaline earth metals and their hydrophobicity that may be related to their buffering capacity and the adsorption of polar and non-polar compounds, such as NH4+ and CO2. It also has aromatic functional groups, just as quinones, whose potential as a redox mediator has been demonstrated and its morphology allows it to form an immobilizing matrix that favors a closer activity among the syntrophic microorganisms, which directly contributed in the oxidation of secondary metabolites and the final reduction to methane, whose yield is increased by 39% compared to controls, with a CBC dose of 1 g/L.

Keywords: anaerobic digestion, biochar, biostimulation, syntrophic metabolism

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193 Recovery and Identification of Phenolic Acids in Honey Samples from Different Floral Sources of Pakistan Having Antimicrobial Activity

Authors: Samiyah Tasleem, Muhammad Abdul Haq, Syed Baqir Shyum Naqvi, Muhammad Abid Husnain, Sajjad Haider Naqvi

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The objective of the present study was: a) to investigate the antimicrobial activity of honey samples of different floral sources of Pakistan, b) to recover the phenolic acids in them as a possible contributing factor of antimicrobial activity. Six honey samples from different floral sources, namely: Trachysperm copticum, Acacia species, Helianthus annuus, Carissa opaca, Zizyphus and Magnifera indica were used. The antimicrobial activity was investigated by the disc diffusion method against eight freshly isolated clinical isolates (Staphylococci aureus, Staphylococci epidermidis, Streptococcus faecalis, Pseudomonas aeruginosa, Klebsiella pneumonia, Escherichia coli, Proteus vulgaris and Candida albicans). Antimicrobial activity of honey was compared with five commercial antibiotics, namely: doxycycline (DO-30ug/mL), oxytetracycline (OT-30ug/mL), clarithromycin (CLR–15ug/mL), moxifloxacin (MXF-5ug/mL) and nystatin (NT – 100 UT). The fractions responsible for antimicrobial activity were extracted using ethyl acetate. Solid phase extraction (SPE) was used to recover the phenolic acids of honey samples. Identification was carried out via High-Performance Liquid Chromatography (HPLC). The results indicated that antimicrobial activity was present in all honey samples and found comparable to the antibiotics used in the study. In the microbiological assay, the ethyl acetate honey extract was found to exhibit a very promising antimicrobial activity against all the microorganisms tested, indicating the existence of phenolic compounds. Six phenolic acids, namely: gallic, caffeic, ferulic, vanillic, benzoic and cinnamic acids were identified besides some unknown substance by HPLC. In conclusion, Pakistani honey samples showed a broad spectrum antibacterial and promising antifungal activity. Identification of six different phenolic acids showed that Pakistani honey samples are rich sources of phenolic compounds that could be the contributing factor of antimicrobial activity.

Keywords: Pakistani honey, antimicrobial activity, Phenolic acids eg.gallic, caffeic, ferulic, vanillic, benzoic and cinnamic acids

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192 Performance and Physiological Responses of Broiler Chickens to Diets Supplemented with Propolis in Breeding, to in Ovo Propolis Feeding or to Propolis Supplementation of Diets for Their Chicks

Authors: Kalbiye Konanc, Ergin Ozturk

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To examine the effects of an ethanol liquid extract obtained from raw bee propolis (PE) on fattening performance and physiology such as vaccine-antibody relationship, microbial profile, immune status and some blood parameters of broiler chickens were used a total of 600 broiler (Ross 308) chicks, obtained from eggs of 288, 38-weeks-old broiler breeding. There were 6 groups: CC (Parent-Control and Offspring-Control, CP (Parent-Control and Offspring-propolis extract, Cip (Parent-Control and Offspring-in-ovo propolis extract), Cis (Parent-Control and Chickens-in-ovo saline), PeC (Parent-propolis extract and Offspring-Control), PeP (Parent-Propolis extract and Offspring-Propolis extract). Each group was consisted of 10 replications with 10 broiler offspring, and the experiment was lasted for 6 weeks with ethanol-extracted propolis concentration is 400 ppm/kg diet. While the highest feed consumptions at 0-21 days and 0-42 days were found in PeC, the best feed conversion ratio at 0-42 days was found in CP group. The live weight gains were found not to be different among the groups. The highest alanine aminotransferase activities were found in CC and CP and aspartate aminotransferase activities in PeP and PeC groups. The highest triglyceride and total antioxidant levels were found highest in CC and the highest total oxidant level in Cip group. IgA level in hatched eggs and IgM value after slaughtering were highest in Cip group. The best immune response was obtained for 21st day Newcastle Disease vaccine in CC and Cis groups and for 28th day Infectious Bursal Disease vaccine in CP group. The highest total aerobic microorganism and the lowest total fungi count were found in PeP group. In conclusion, it was determined that in-ovo propolis ethanol extract (Cip) increased the maternal antibody levels, that had not consistent effects on blood biochemical parameters except for triglyceride, that led to decrease in E. coli counts and that it can provide strong immune response against Infectious Bursal Disease.

Keywords: bee propolis, in-ovo feeding, immune parameters, poultry, maternal antibody, microorganisms

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191 Mitigating Biofouling on Reverse Osmosis Membranes: Applying Greener Preservatives to Biofilm Treatment

Authors: Anna Curtin, Matthew Thibodeau, Heather Buckley

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Water scarcity is characterized by a lack of access to clean and affordable drinking water, as well as water for hygienic and economic needs. The amount of people effected by water scarcity is expected to increase in the coming years due to climate change, population growth, and pollution, amongst other things. In response, scientists are pursuing cost effective drinking water treatment methods, often with a focus on alternative water sources. Desalination of seawater via reverse osmosis is one promising alternative method. Desalination of seawater via reverse osmosis, however, is limited significantly by biofouling of the filtration membrane. Biofouling is the buildup of microorganisms in a biofilm at the water-membrane interface. It clogs the membrane, decreasing the efficiency of filtration, consequently increasing operational and maintenance costs. Although effective, existing chemical treatment methods can damage the membrane, decreasing the lifespan of the membrane; create antibiotic resistance; and cause harm to humans and the environment if they pass through the membrane into the permeate. The current project focuses on applying safer preservatives used in home and personal care products to RO membranes to investigate the biofouling treatment efficacy. Currently, many of these safer preservatives have only been tested on cells in planktonic phase in suspension cultures, not on cells in biofilms. The results of suspension culture tests are not applicable to biofouling scenarios because organisms in planktonic phase in suspension cultures exhibit different morphological, chemical, and metabolic characteristics than those in a biofilm. Testing antifoulant efficacy of safer preservatives on biofilms will provide more applicable results to biofouling on RO membranes. To do this, biofilms will be grown on 96-well-plates and minimum inhibitory concentrations (MIC90) and log-reductions will be calculated for various safer preservatives. Results from these tests will be used to guide doses for tests of safer preservatives in a bench-scale RO system.

Keywords: reverse osmosis, biofouling, preservatives, antimicrobial, safer alternative, green chemistry

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190 Bioefficiency of Cinnamomum verum Loaded Niosomes and Its Microbicidal and Mosquito Larvicidal Activity against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus

Authors: Aasaithambi Kalaiselvi, Michael Gabriel Paulraj, Ekambaram Nakkeeran

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Emergences of mosquito vector-borne diseases are considered as a perpetual problem globally in tropical countries. The outbreak of several diseases such as chikungunya, zika virus infection and dengue fever has created a massive threat towards the living population. Frequent usage of synthetic insecticides like Dichloro Diphenyl Trichloroethane (DDT) eventually had its adverse harmful effects on humans as well as the environment. Since there are no perennial vaccines, prevention, treatment or drugs available for these pathogenic vectors, WHO is more concerned in eradicating their breeding sites effectively without any side effects on humans and environment by approaching plant-derived natural eco-friendly bio-insecticides. The aim of this study is to investigate the larvicidal potency of Cinnamomum verum essential oil (CEO) loaded niosomes. Cholesterol and surfactant variants of Span 20, 60 and 80 were used in synthesizing CEO loaded niosomes using Transmembrane pH gradient method. The synthesized CEO loaded niosomes were characterized by Zeta potential, particle size, Fourier Transform Infrared Spectroscopy (FT-IR), GC-MS and SEM analysis to evaluate charge, size, functional properties, the composition of secondary metabolites and morphology. The Z-average size of the formed niosomes was 1870.84 nm and had good stability with zeta potential -85.3 meV. The entrapment efficiency of the CEO loaded niosomes was determined by UV-Visible Spectrophotometry. The bio-potency of CEO loaded niosomes was treated and assessed against gram-positive (Bacillus subtilis) and gram-negative (Escherichia coli) bacteria and fungi (Aspergillus fumigatus and Candida albicans) at various concentrations. The larvicidal activity was evaluated against II to IV instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus at various concentrations for 24 h. The mortality rate of LC₅₀ and LC₉₀ values were calculated. The results exhibited that CEO loaded niosomes have greater efficiency against mosquito larvicidal activity. The results suggest that niosomes could be used in various applications of biotechnology and drug delivery systems with greater stability by altering the drug of interest.

Keywords: Cinnamomum verum, niosomes, entrapment efficiency, bactericidal and fungicidal, mosquito larvicidal activity

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189 The Comparison Study of Human Microbiome in Chronic Rhinosinusitis between Adults and Children

Authors: Il Ho Park, Joong Seob Lee, Sung Hun Kang, Jae-Min Shin, Il Seok Park, Seok Min Hong, Seok Jin Hong

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Introduction: The human microbiota is the aggregate of microorganisms, and the bacterial microbiome of the human digestive tract contributes to both health and disease. In health, bacteria are key components in the development of mucosal barrier function and in innate and adaptive immune responses, and they also work to suppress the establishment of pathogens. In human upper airway, the sinonasal microbiota might play an important role in chronic rhinosinusitis (CRS). The purpose of this study is to investigate the human upper airway microbiome in CRS patients and to compare the sinonasal microbiome of adults with children. Materials and methods: A total of 19 samples from 19 patients (Group1; 9 CRS in children, aged 5 to 14 years versus Group 2; 10 CRS in adults aged 21 to 59 years) were examined. Swabs were collected from the middle meatus and/or anterior ethmoid region under general anesthesia during endoscopic sinus surgery or tonsillectomy. After DNA extraction from swab samples, we analysed bacterial microbiome consortia using 16s rRNA gene sequencing approach (the Illumina MiSeq platform). Results: In this study, relatively abundance of the six bacterial phyla and tremendous genus and species found in substantial amounts in the individual sinus swab samples, include Corynebacterium, Hemophilus, Moraxella, and Streptococcus species. Anaerobes like Fusobacterium and Bacteroides were abundantly present in the children group, Bacteroides and Propionibacterium were present in adults group. In genus, Haemophilus was the most common CRS microbiome in children and Corynebacterium was the most common CRS microbiome in adults. Conclusions: Our results show the diversity of human upper airway microbiome, and the findings will suggest that CRS is a polymicrobial infection. The Corynebacterium and Hemophilus may live as commensals on mucosal surfaces of sinus in the upper respiratory tract. The further study will be needed for analysis of microbiome-human interactions in upper airway and CRS.

Keywords: microbiome, upper airway, chronic rhinosinusitis, adult and children

Procedia PDF Downloads 99
188 Biological Hazards and Laboratory inflicted Infections in Sub-Saharan Africa

Authors: Godfrey Muiya Mukala

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This research looks at an array of fields in Sub-Saharan Africa comprising agriculture, food enterprises, medicine, organisms genetically modified, microbiology, and nanotechnology that can be gained from biotechnological research and development. Findings into dangerous organisms, mainly bacterial germs, rickettsia, fungi, parasites, or organisms that are genetically engineered, have immensely posed questions attributed to the biological danger they bring forth to human beings and the environment because of their uncertainties. In addition, the recurrence of previously managed diseases or the inception of new diseases are connected to biosafety challenges, especially in rural set-ups in low and middle-income countries. Notably, biotechnology laboratories are required to adopt biosafety measures to protect their workforce, community, environment, and ecosystem from unforeseen materials and organisms. Sensitization and inclusion of educational frameworks for laboratory workers are essential to acquiring a solid knowledge of harmful biological agents. This is in addition to human pathogenicity, susceptibility, and epidemiology to the biological data used in research and development. This article reviews and analyzes research intending to identify the proper implementation of universally accepted practices in laboratory safety and biological hazards. This research identifies ideal microbiological methods, adequate containment equipment, sufficient resources, safety barriers, specific training, and education of the laboratory workforce to decrease and contain biological hazards. Subsequently, knowledge of standardized microbiological techniques and processes, in addition to the employment of containment facilities, protective barriers, and equipment, is far-reaching in preventing occupational infections. Similarly, reduction of risks and prevention may be attained by training, education, and research on biohazards, pathogenicity, and epidemiology of the relevant microorganisms. In this technique, medical professionals in rural setups may adopt the knowledge acquired from the past to project possible concerns in the future.

Keywords: sub-saharan africa, biotechnology, laboratory, infections, health

Procedia PDF Downloads 59
187 Nanopack: A Nanotechnology-Based Antimicrobial Packaging Solution for Extension of Shelf Life and Food Safety

Authors: Andy Sand, Naama Massad – Ivanir, Nadav Nitzan, Elisa Valderrama, Alfred Wegenberger, Koranit Shlosman, Rotem Shemesh, Ester Segal

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Microbial spoilage of food products is of great concern in the food industry due to the direct impact on the shelf life of foods and the risk of foodborne illness. Therefore, food packaging may serve as a crucial contribution to keep the food fresh and suitable for consumption. Active packaging solutions that have the ability to inhibit the development of microorganism in food products attract a lot of interest, and many efforts have been made to engineer and assimilate such solutions on various food products. NanoPack is an EU-funded international project aiming to develop state-of-the-art antimicrobial packaging systems for perishable foods. The project is based on natural essential oils which possess significant antimicrobial activity against many bacteria, yeasts and molds. The essential oils are encapsulated in natural aluminosilicate clays, halloysite nanotubes (HNT's), that serves as a carrier for the volatile essential oils and enable their incorporation into polymer films. During the course of the project, several polyethylene films with diverse essential oils combinations were designed based on the characteristics of their target food products. The antimicrobial activity of the produced films was examined in vitro on a broad spectrum of microorganisms including gram-positive and gram-negative bacteria, aerobic and anaerobic bacteria, yeasts and molds. The films that showed promising in vitro results were successfully assimilated on in vivo active packaging of several food products such as cheese, bread, fruits and raw meat. The results of the in vivo analyses showed significant inhibition of the microbial spoilage, indicating the strong contribution of the NanoPack packaging solutions on the extension of shelf life and reduction of food waste caused by early spoilage throughout the supply chain.

Keywords: food safety, food packaging, essential oils, nanotechnology

Procedia PDF Downloads 113
186 Characterization and Optimization of Culture Conditions for Sulphur Oxidizing Bacteria after Isolation from Rhizospheric Mustard Soil, Decomposing Sites and Pit House

Authors: Suman Chaudhary, Rinku Dhanker, Tanvi, Sneh Goyal

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Sulphur oxidizing bacteria (SOB) have marked their significant role in perspectives of maintaining healthy environment as researchers from all over the world tested and apply these in waste water treatment plants, bioleaching of heavy metals, deterioration of bridge structures, concrete and for bioremediation purposes, etc. Also, these SOB are well adapted in all kinds of environment ranging from normal soil, water habitats to extreme natural sources like geothermal areas, volcanic eruptions, black shale and acid rock drainage (ARD). SOB have been isolated from low pH environment of anthropogenic origin like acid mine drainage (AMD) and bioleaching heaps, hence these can work efficiently in different environmental conditions. Besides having many applications in field of environment science, they may be proven to be very beneficial in area of agriculture as sulphur is the fourth major macronutrients required for the growth of plants. More amount of sulphur is needed by pulses and oilseed crops with respect to the cereal grains. Due to continuous use of land for overproduction of more demanding sulphur utilizing crops and without application of sulphur fertilizers, its concentration is decreasing day by day, and thus, sulphur deficiency is becoming a great problem as it affects the crop productivity and quality. Sulphur is generally found in soils in many forms which are unavailable for plants (cannot be use by plants) like elemental sulphur, thiosulphate which can be taken up by bacteria and converted into simpler forms usable by plants by undergoing a series of transformations. So, keeping the importance of sulphur in view for various soil types, oilseed crops and role of microorganisms in making them available to plants, we made an effort to isolate, optimize, and characterize SOB. Three potential strains of bacteria were isolated, namely SSF7, SSA21, and SSS6, showing sulphate production of concentration, i.e. 2.268, 3.102, and 2.785 mM, respectively. Also, these were optimized for various culture conditions like carbon, nitrogen source, pH, temperature, and incubation time, and characterization was also done.

Keywords: sulphur oxidizing bacteria, isolation, optimization, characterization, sulphate production

Procedia PDF Downloads 318
185 Enzymatic Degradation of Poly (Butylene Adipate Terephthalate) Copolymer Using Lipase B From Candida Antarctica and Effect of Poly (Butylene Adipate Terephthalate) on Plant Growth

Authors: Aqsa Kanwal, Min Zhang, Faisal Sharaf, Li Chengtao

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The globe is facing increasing challenges of plastic pollution due to single-use of plastic-based packaging material. The plastic material is continuously being dumped into the natural environment, which causes serious harm to the entire ecosystem. Polymer degradation in nature is very difficult, so the use of biodegradable polymers instead of conventional polymers can mitigate this issue. Due to the good mechanical properties and biodegradability, aliphatic-aromatic polymers are being widely commercialized. Due to the advancement in molecular biology, many studies have reported specific microbes that can effectively degrade PBAT. Aliphatic polyesters undergo hydrolytic cleavage of ester groups, so they can be easily degraded by microorganisms. In this study, we investigated the enzymatic degradation of poly (butylene adipate terephthalate) (PBAT) copolymer using lipase B from Candida Antarctica (CALB). Results of the study displayed approximately 5.16 % loss in PBAT mass after 2 days which significantly increased to approximately 15.7 % at the end of the experiment (12 days) as compared to blank. The pH of the degradation solution also displayed significant reduction and reached the minimum value of 6.85 at the end of the experiment. The structure and morphology of PBAT after degradation were characterized by FTIR, XRD, SEM, and TGA. FTIR analysis showed that after degradation many peaks become weaker and the peak at 2950 cm-1 almost disappeared after 12 days. The XRD results indicated that as the degradation time increases the intensity of diffraction peaks slightly increases as compared to the blank PBAT. TGA analysis also confirmed the successful degradation of PBAT with time. SEM micrographs further confirmed that degradation has occurred. Hence, biodegradable polymers can widely be used. The effect of PBAT biodegradation on plant growth was also studied and it was found that PBAT has no toxic effect on the growth of plants. Hence PBAT can be employed in a wide range of applications.

Keywords: aliphatic-aromatic co-polyesters, polybutylene adipate terephthalate, lipase (CALB), biodegradation, plant growth

Procedia PDF Downloads 57
184 Microalgae for Plant Biostimulants on Whey and Dairy Wastewaters

Authors: Sergejs Kolesovs, Pavels Semjonovs

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Whey and dairy wastewaters if disposed in the environment without proper treatment, cause serious environmental risks contributing to overall and particular environmental pollution and climate change. Biological treatment of wastewater is considered to be most eco-friendly approach, as compared to the chemical treatment methods. Research shows, that dairy wastewater can potentially be remediated by use of microalgae thussignificantly reducing the content of carbohydrates, P, N, K and other pollutants. Moreover, it has been shown, that use of dairy wastewaters results in higher microalgae biomass production. In recent decades microalgal biomass has entailed a big interest for its potential applications in pharmaceuticals, biomedicine, health supplementation, cosmetics, animal feed, plant protection, bioremediation and biofuels. It was shown, that lipids productivity on whey and dairy wastewater is higher as compared with standard cultivation media and occurred without the necessity of inducing specific stress conditions such as N starvation. Moreover, microalgae biomass production as usually associated with high production costs may benefit from perspective of both reasons – enhanced microalgae biomass or target substances productivity on cheap growth substrate and effective management of whey and dairy wastewaters, which issignificant for decrease of total production costs in both processes. Obviously, it became especially important when large volume and low cost industrial microalgal biomass production is anticipated for further use in agriculture of crops as plant growth stimulants, biopesticides soil fertilisers or remediating solutions. Environmental load of dairy wastewaters can be significantly decreased when microalgae are grown in coculture with other microorganisms. This enhances the utilisation of lactose, which is main C source in whey and dairy wastewaters when it is not metabolised easily by most microalgal species chosen. Our study showsthat certain microalgae strains can be used in treatment of residual sugars containing industrial wastewaters and decrease of their concentration thus approving that further extensive research on dairy wastewaters pre-treatment optionsfor effective cultivation of microalgae, carbon uptake and metabolism, strain selection and choice of coculture candidates is needed for further optimisation of the process.

Keywords: microalgae, whey, dairy wastewaters, sustainability, plant biostimulants

Procedia PDF Downloads 71
183 Delineation of Oil– Polluted Sites in Ibeno LGA, Nigeria

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

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Ibeno, Nigeria hosts the operational base of Mobil Producing Nigeria Unlimited (MPNU), a subsidiary of ExxonMobil and the current highest oil and condensate producer in Nigeria. Besides MPNU, other multinational oil companies like Shell Petroleum Development Company Ltd, Elf Petroleum Nigeria Ltd and Nigerian Agip Energy, a subsidiary of ENI E&P operate onshore, on the continental shelf and deep offshore of the Atlantic Ocean in Ibeno, Nigeria, respectively. This study was designed to carry out the survey of the oil impacted sites in Ibeno, Nigeria. A combinations of electrical resistivity (ER), ground penetrating radar (GPR) and physico-chemical as well as microbiological characterization of soils and water samples from the area were carried out. Results obtained revealed that there have been hydrocarbon contaminations of this environment by past crude oil spills as observed from significant concentrations of THC, BTEX and heavy metal contents in the environment. Also, high resistivity values and GPR profiles clearly showing the distribution, thickness and lateral extent of hydrocarbon contamination as represented on the radargram reflector tones corroborates previous significant oil input. Contaminations were of varying degrees, ranging from slight to high, indicating levels of substantial attenuation of crude oil contamination over time. Hydrocarbon pollution of the study area was confirmed by the results of soil and water physico-chemical and microbiological analysis. The levels of THC contamination observed in this study are indicative of high levels of crude oil contamination. Moreover, the display of relatively lower resistivities of locations outside the impacted areas compared to resistivity values within the impacted areas, the 3-D Cartesian images of oil contaminant plume depicted by red, light brown and magenta for high, low and very low oil impacted areas, respectively as well as the high counts of hydrocarbonoclastic microorganisms in excess of 1% confirmed significant recent pollution of the study area.

Keywords: oil-polluted sites, physico-chemical analyses, microbiological characterization, geotechnical investigations, total hydrocarbon content

Procedia PDF Downloads 373
182 Peculiarities of Microflora of Odontogenic Inflammatory Processes in the Central Kazakhstan Region

Authors: Aliya Tokbergenova, Maida Tusupbekova, Daulet Dzhangaliyev, Alena Lavrinenko

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Background: Odontogenic phlegmons are ranked the first among pyoinflammatory processes in the frequency of hospitalization in maxillofacial surgery in the post-Soviet countries. The main role in etiology is played by obligate anaerobes and aerobes. According to numerous data, the structure of aerobic pathogens is dominated by staphylococci and gram-negative bacteria. Aim: The research aim is to study the microflora of the purulent discharge odontogenic inflammatory processes. Materials and methods: A total of 220 patients have been examined, of which 120 patients aged 25-59 years have been included in the research who did not have comorbidity hospitalized in the maxillofacial hospital in Karaganda (Kazakhstan) from January 2016 to July 2017. The bacteriological research has been carried out on the basis of the multiaccess laboratory of the KSMU, through the Matrix Assisted Laser Desorption/Ionization (MALDI) apparatus. The material sample was pus from the inflammation focus, taken during the operating period. Results: According to the research among 120 patients (100%), 15 patients (12.5%) have had microorganisms not grown. From 105 (87.5%) bacteriological results, it has been revealed the following 1) Streptococcus: 51 (42.5%): Streptococcus beta-haemolytic: 17 (14.2%), Streptococcus pneumoniae: 12 (10%), Streptococcus anginosus: 8 (6.6%), Streptococcus oralis: 8 (6.6%), Streptococcus constellatus: 6 (5.0%); 2) Staphylococci: 27 (22.5%): Staphylococci aureus: 14 (11.7%) and Staphylococci epidermidis: 13 (10.8%); 3) Pseudomonas aeruginosa: 12 (10%); 4) Neisseria: 11 (9.1%): Neisseria mucosa: 5 (4.1%) and Neisseria macacae: 6 (5.0%); 5) Klebsiella pneumoniae: 2 (1.7%); 6) Stenotrophomonas maltophilia: 2 (1.7%). 15 patients (12.5%) experienced complications in the form of 1) The dissemination of the process in 10 patients (8.4%). 2) Osteomyelitis in 3 (2.5%). 3) Mediastinitis in 1 (0.8%). 4) Sinusitis in 1 (0.8%). 15 patients (100%) were carried out repeated bacteriological examination, the following was revealed: 1) Streptococcus: 10 (66.7%): Streptococcus beta-haemolytic: 4 (26.7%), Streptococcus pneumoniae: 2 (13.3%), Streptococcus аnginosus: 2 (13.3%), Streptococcus oralis: 1 (6.7%), Streptococcus constellatus: 1 (6.7%); 2) Staphylococci: 4 (26.7%): Staphylococci aureus: 3 (20%) and Staphylococci epidermidis: 1 (6.7%); 3) Pseudomonas aeruginosa: 1 (6.7%). Conclusions: Thus, according to our research data, streptococci predominate in the odontogenic processes microflora in aerobic flora in the central Kazakhstan region, which refutes the leading role of staphylococci in the development of odontogenic inflammatory processes, thus creating prerequisites for studying new treatment approaches.

Keywords: maxillofacial surgery, microflora, odontogenic phlegmons, pyo-inflammatory

Procedia PDF Downloads 160
181 Microbial Effects of Iron Elution from Hematite into Seawater Mediated via Dissolved Organic Matter

Authors: Apichaya Aneksampant, Xuefei Tu, Masami Fukushima, Mitsuo Yamamoto

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The restoration of seaweed beds recovery has been developed using a fertilization technique for supplying dissolved iron to barren coastal areas. The fertilizer is composed of iron oxides as a source of iron and compost as humic substance (HS) source, which can serve as chelator of iron to stabilize the dissolved species under oxic seawater condition. However, elution mechanisms of iron from iron oxide surfaces have not sufficiently elucidated. In particular, roles of microbial activities in the elution of iron from the fertilizer are not sufficiently understood. In the present study, a fertilizer (iron oxide/compost = 1/1, v/v) was incubated in a water tank at Mashike coast, Hokkaido Japan. Microorganisms in the 6-month fertilizer were isolated and identified as Exiguobacterium oxidotolerans sp. (T-2-2). The identified bacteria were inoculated to perform iron elution test in a postgate B medium, prepared in artificial seawater. Hematite was used as a model iron oxide and anthraquinone-2,7-disolfonate (AQDS) as a model for HSs. The elution test performed in presence and absence of bacteria inoculation. ICP-AES was used to analyze total iron and a colorimetric technique using ferrozine employed for the determination of ferrous ion. During the incubation period, sample contained hematite and T-2-2 in both presence and absence of AQDS continuously showed the iron elution and reached at the highest concentration after 9 days of incubation and then slightly decrease to stabilize within 20 days. Comparison to the sample without T-2-2, trace amount of iron was observed, suggesting that iron elution to seawater can be attributed to bacterial activities. The levels of total organic carbon (TOC) in the culture solution with hematite decreased. This may be to the adsorption of organic compound, AQDS, to hematite surfaces. The decrease in UV-vis absorption of AQDS in the culture solution also support the results of TOC that AQDS was adsorbed to hematite surfaces. AQDS can enhance the iron elution, while the adsorption of organic matter suppresses the iron elution from hematite.

Keywords: anthraquinone-2, 7-disolfonate, barren ground, E.oxidotolerans sp., hematite, humic substances, iron elution

Procedia PDF Downloads 356
180 Immuno-Modulatory Role of Weeds in Feeds of Cyprinus Carpio

Authors: Vipin Kumar Verma, Neeta Sehgal, Om Prakash

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Cyprinus carpio has a wide spread occurrence in the lakes and rivers of Europe and Asia. Heavy losses in natural environment due to anthropogenic activities, including pollution as well as pathogenic diseases have landed this fish in IUCN red list of vulnerable species. The significance of a suitable diet in preserving the health status of fish is widely recognized. In present study, artificial feed supplemented with leaves of two weed plants, Eichhornia crassipes and Ricinus communis were evaluated for their role on the fish immune system. To achieve this objective fish were acclimatized to laboratory conditions (25 ± 1 °C; 12 L: 12D) for 10 days prior to start of experiment and divided into 4 groups: non-challenged (negative control= A), challenged [positive control (B) and experimental (C & D)]. Group A, B were fed with non-supplemented feed while group C & D were fed with feed supplemented with 5% Eichhornia crassipes and 5% Ricinus communis respectively. Supplemented feeds were evaluated for their effect on growth, health, immune system and disease resistance in fish when challenged with Vibrio harveyi. Fingerlings of C. carpio (weight, 2.0±0.5 g) were exposed with fresh overnight culture of V. harveyi through bath immunization (concentration 2 Χ 105) for 2 hours on 10 days interval for 40 days. The growth was monitored through increase in their relative weight. The rate of mortality due to bacterial infection as well as due to effect of feed was recorded accordingly. Immune response of fish was analyzed through differential leucocyte count, percentage phagocytosis and phagocytic index. The effect of V. harveyi on fish organs were examined through histo-pathological examination of internal organs like spleen, liver and kidney. The change in the immune response was also observed through gene expression analysis. The antioxidant potential of plant extracts was measured through DPPH and FRAP assay and amount of total phenols and flavonoids were calculates through biochemical analysis. The chemical composition of plant’s methanol extracts was determined by GC-MS analysis, which showed presence of various secondary metabolites and other compounds. Investigation revealed immuno-modulatory effect of plants, when supplemented with the artificial feed of fish.

Keywords: immuno-modulation, gc-ms, Cyprinus carpio, Eichhornia crassipes, Ricinus communis

Procedia PDF Downloads 460
179 Isolation, Identification and Screening of Marine Fungi for Potential Tyrosinase Inhibitor, Antibacterial and Antioxidant for Future Cosmeceuticals

Authors: Shivankar Agrawal, Sunil Kumar Deshmukh, Colin Barrow, Alok Adholeya

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A variety of genetic and environmental factors cause various cosmetics and dermatological problems. There are already claimed drugs available in market for treating these problems. However, the challenge remains in finding more potent, environmental friendly, causing minimal side effects and economical cosmeceuticals. This leads to an increased demand for natural cosmeceutical products in the last few decades. Plant derived ingredients are limited because plants either contain toxic metabolites, grow too slow or seasonal harvesting is a problem. To identify new bioactive cosmetics ingredients of marine microbial bioresource, we screened 35 marine fungi isolated from marine samples collected from Andaman Island and west coast of India. Fungal crude extracts were investigated for their antityrosinase, antioxidant and antibacterial activities for the purpose of identifying anti-aging, skin-whitening and anti-acne biomolecule with the potential in cosmetics. In the tyrosinase inhibition and 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assays, two fungal extracts, including “P2”, Talaromyces stipitatus and “D4”, Aspergillus terreus showed high inhibitory activity at 1mg/mL for tyrosinase inhibition and 0.5mg/mL for DPPH scavenging. The in vitro antimicrobial activity was investigated by the agar well diffusion method. In the tyrosinase inhibition assay, 8 extracts showed significant antibacterial activity against bacteria causing skin and wound infection in humans. In the course of systematic screening program for bioactive marine fungi, strain “D5” was found to be most potent strain with MIC value of 1mg/mL, which was morphologically identified as Simplicillium lamellicola. The effects of the most active crude extracts against their susceptible test microorganisms were also investigated by SEM analysis. Further investigations will focus on purification and characterization major active components responsible for these activities.

Keywords: antioxidant, antimicrobial activity, tyrosinase, cosmeceuticals, marine fungi

Procedia PDF Downloads 258
178 Postmortem Genetic Testing to Sudden and Unexpected Deaths Using the Next Generation Sequencing

Authors: Eriko Ochiai, Fumiko Satoh, Keiko Miyashita, Yu Kakimoto, Motoki Osawa

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Sudden and unexpected deaths from unknown causes occur in infants and youths. Recently, molecular links between a part of these deaths and several genetic diseases are examined in the postmortem. For instance, hereditary long QT syndrome and Burgada syndrome are occasionally fatal through critical ventricular tachyarrhythmia. There are a large number of target genes responsible for such diseases, the conventional analysis using the Sanger’s method has been laborious. In this report, we attempted to analyze sudden deaths comprehensively using the next generation sequencing (NGS) technique. Multiplex PCR to subject’s DNA was performed using Ion AmpliSeq Library Kits 2.0 and Ion AmpliSeq Inherited Disease Panel (Life Technologies). After the library was constructed by emulsion PCR, the amplicons were sequenced 500 flows on Ion Personal Genome Machine System (Life Technologies) according to the manufacture instruction. SNPs and indels were analyzed to the sequence reads that were mapped on hg19 of reference sequences. This project has been approved by the ethical committee of Tokai University School of Medicine. As a representative case, the molecular analysis to a 40 years old male who received a diagnosis of Brugada syndrome demonstrated a total of 584 SNPs or indels. Non-synonymous and frameshift nucleotide substitutions were selected in the coding region of heart disease related genes of ANK2, AKAP9, CACNA1C, DSC2, KCNQ1, MYLK, SCN1B, and STARD3. In particular, c.629T-C transition in exon 3 of the SCN1B gene, resulting in a leu210-to-pro (L210P) substitution is predicted “damaging” by the SIFT program. Because the mutation has not been reported, it was unclear if the substitution was pathogenic. Sudden death that failed in determining the cause of death constitutes one of the most important unsolved subjects in forensic pathology. The Ion AmpliSeq Inherited Disease Panel can amplify the exons of 328 genes at one time. We realized the difficulty in selection of the true source from a number of candidates, but postmortem genetic testing using NGS analysis deserves of a diagnostic to date. We now extend this analysis to SIDS suspected subjects and young sudden death victims.

Keywords: postmortem genetic testing, sudden death, SIDS, next generation sequencing

Procedia PDF Downloads 333
177 Insights into the Annotated Genome Sequence of Defluviitoga tunisiensis L3 Isolated from a Thermophilic Rural Biogas Producing Plant

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

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

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

Procedia PDF Downloads 471
176 Data Analysis for Taxonomy Prediction and Annotation of 16S rRNA Gene Sequences from Metagenome Data

Authors: Suchithra V., Shreedhanya, Kavya Menon, Vidya Niranjan

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Skin metagenomics has a wide range of applications with direct relevance to the health of the organism. It gives us insight to the diverse community of microorganisms (the microbiome) harbored on the skin. In the recent years, it has become increasingly apparent that the interaction between skin microbiome and the human body plays a prominent role in immune system development, cancer development, disease pathology, and many other biological implications. Next Generation Sequencing has led to faster and better understanding of environmental organisms and their mutual interactions. This project is studying the human skin microbiome of different individuals having varied skin conditions. Bacterial 16S rRNA data of skin microbiome is downloaded from SRA toolkit provided by NCBI to perform metagenomics analysis. Twelve samples are selected with two controls, and 3 different categories, i.e., sex (male/female), skin type (moist/intermittently moist/sebaceous) and occlusion (occluded/intermittently occluded/exposed). Quality of the data is increased using Cutadapt, and its analysis is done using FastQC. USearch, a tool used to analyze an NGS data, provides a suitable platform to obtain taxonomy classification and abundance of bacteria from the metagenome data. The statistical tool used for analyzing the USearch result is METAGENassist. The results revealed that the top three abundant organisms found were: Prevotella, Corynebacterium, and Anaerococcus. Prevotella is known to be an infectious bacterium found on wound, tooth cavity, etc. Corynebacterium and Anaerococcus are opportunist bacteria responsible for skin odor. This result infers that Prevotella thrives easily in sebaceous skin conditions. Therefore it is better to undergo intermittently occluded treatment such as applying ointments, creams, etc. to treat wound for sebaceous skin type. Exposing the wound should be avoided as it leads to an increase in Prevotella abundance. Moist skin type individuals can opt for occluded or intermittently occluded treatment as they have shown to decrease the abundance of bacteria during treatment.

Keywords: bacterial 16S rRNA , next generation sequencing, skin metagenomics, skin microbiome, taxonomy

Procedia PDF Downloads 151
175 Dynamics of Bacterial Contamination and Oral Health Risks Associated with Currency Notes and Coins Circulating in Kampala City

Authors: Abdul Walusansa

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In this paper, paper notes and coins were collected from general public in Kampala City where ready-to-eat food can be served, in order to survey for bacterial contamination. The total bacterial number and potentially pathogenic organisms loading on currency were tested. All isolated potential pathogens were also tested for antibiotic resistance against four most commonly prescribed antibiotics. 1. The bacterial counts on one hundred paper notes sample were ranging between 6~10918/cm cm-2,the median was 141/ cm-2, according to the data it was much higher than credit cards and Australian notes which were made of polymer. The bacterial counts on sixty coin samples were ranging between 2~380/cm-2, much less than paper notes. 2. Coliform (65.6%), E. coli (45.9%), S. aureus (41.7%), B. cereus (67.7%), Salmonella (19.8%) were isolated on one hundred paper notes. Coliform (22.4%), E. coli (5.2%), S. aureus (24.1%), B. cereus (34.5%), Salmonella (10.3%) were isolated from sixty coin samples. These results suggested a high rate of potential pathogens contamination of paper notes than coins. 3. Antibiotic resistances are commonly in most of the pathogens isolated on currency. Ampicillin resistance was found in 60%of Staphylococcus aureus isolated on currency, as well as 76.6% of E. coil and 40% of Salmonella. Erythromycin resistance was detected in 56.6% of S. aureus and in 80.0% of E. coli. All the pathogens isolated were sensitive to Norfloxacin, Salmonella and S. aureus also sensitive to Cefaclor. In this paper, we also studied the antimicrobial capability of metal coins, coins collected from different countries were tested for the ability to inhibit the growth of E. sakazakii, S. aureus, E. coli, L. monocytogenes and S. typhimurium. 1) E. sakazakii appeared very sensitive to metal coins, the second is S. aureus, but E. coli, L. monocytogenes and S. typhimurium are more resistant to these metal coin samples. 2) Coins made of Nickel-brass alloy and Copper-nickel alloy showed a better effect in anti-microbe than other metal coins, especially the ability to inhibited the growth of E. sakazakii and S. aureus, all the inhibition zones produced on nutrient agar are more than 20.6 mm. Aluminium-bronze alloy revealed weak anti-microbe activity to S. aureus and no effect to kill other pathogens. Coins made of stainless steel also can’t resist bacteria growth. 3) Surprisingly, one cent coins of USA which were made of 97.5% Zinc and 2.5% Cu showed a significant antimicrobial capability, the average inhibition zone of these five pathogens is 45.5 mm.

Keywords: antibiotic sensitivity, bacteria, currency, coins, parasites

Procedia PDF Downloads 302
174 The Dependency of the Solar Based Disinfection on the Microbial Quality of the Source Water

Authors: M. T. Amina, A. A. Alazba, U. Manzoor

Abstract:

Solar disinfection (SODIS) is a viable method for household water treatment and is recommended by the World Health Organization as cost effective approach that can be used without special skills. The efficiency of both SODIS and solar collector disinfection (SOCODIS) system was evaluated using four different sources of water including stored rainwater, storm water, ground water and treated sewage. Samples with naturally occurring microorganisms were exposed to sunlight for about 8-9 hours in 2-L polyethylene terephthalate bottles under similar experimental conditions. Total coliform (TC), Escherichia coli (E. coli) and heterotrophic plate counts (HPC) were used as microbial water quality indicators for evaluating the disinfection efficiency at different sunlight intensities categorized as weak, mild and strong weathers. Heterotrophic bacteria showed lower inactivation rates compared to E. coli and TC in both SODIS and SOCODIS system. The SOCODIS system at strong weather was the strongest disinfection system in this study and the complete inactivation of HPC was observed after 8-9 hours of exposure with SODIS being ineffective for HPC. At moderate weathers, however, the SOCODIS system did not show complete inactivation of HPC due to very high concentrations (up to 5x10^7 CFU/ml) in both storm water and treated sewage. SODIS even remained ineffective for the complete inactivation of E. coli due to its high concentrations of about 2.5x10^5 in treated sewage compared with other waters even after 8-9 hours of exposure. At weak weather, SODIS was not effective at all while SOCODIS system, though incomplete, showed good disinfection efficiency except for HPC and to some extent for high E. coli concentrations in storm water. Largest reduction of >5 log occurred for TC when used stored rainwater even after 6 hours of exposure in the case of SOCODIS system at strong weather. The lowest E. coli and HPC reduction of ~2 log was observed in SODIS system at weak weather. Further tests with varying pH and turbidity are required to understand the effects of reaction parameters that could be a step forward towards maximizing the disinfection efficiency of such systems for the complete inactivation of naturally occurring E. coli or HPC at moderate or even at weak weathers.

Keywords: efficiency, microbial, SODIS, SOCODIS, weathers

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