Search results for: bacterial aggregation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1411

Search results for: bacterial aggregation

271 Standardization of a Methodology for Quantification of Antimicrobials Used for the Treatment of Multi-Resistant Bacteria Using Two Types of Biosensors and Production of Anti-Antimicrobial Antibodies

Authors: Garzon V., Bustos R., Salvador J. P., Marco M. P., Pinacho D. G.

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Bacterial resistance to antimicrobial treatment has increased significantly in recent years, making it a public health problem. Large numbers of bacteria are resistant to all or nearly all known antimicrobials, creating the need for the development of new types of antimicrobials or the use of “last line” antimicrobial drug therapies for the treatment of multi-resistant bacteria. Some of the chemical groups of antimicrobials most used for the treatment of infections caused by multiresistant bacteria in the clinic are Glycopeptide (Vancomycin), Polymyxin (Colistin), Lipopeptide (Daptomycin) and Carbapenem (Meropenem). Molecules that require therapeutic drug monitoring (TDM). Due to the above, a methodology based on nanobiotechnology based on an optical and electrochemical biosensor is being developed, which allows the evaluation of the plasmatic levels of some antimicrobials such as glycopeptide, polymyxin, lipopeptide and carbapenem quickly, at a low cost, with a high specificity and sensitivity and that can be implemented in the future in public and private health hospitals. For this, the project was divided into five steps i) Design of specific anti-drug antibodies, produced in rabbits for each of the types of antimicrobials, evaluating the results by means of an immunoassay analysis (ELISA); ii) quantification by means of an electrochemical biosensor that allows quantification with high sensitivity and selectivity of the reference antimicrobials; iii) Comparison of antimicrobial quantification with an optical type biosensor; iv) Validation of the methodologies used with biosensor by means of an immunoassay. Finding as a result that it is possible to quantify antibiotics by means of the optical and electrochemical biosensor at concentrations on average of 1,000ng/mL, the antibodies being sensitive and specific for each of the antibiotic molecules, results that were compared with immunoassays and HPLC chromatography. Thus, contributing to the safe use of these drugs commonly used in clinical practice and new antimicrobial drugs.

Keywords: antibiotics, electrochemical biosensor, optical biosensor, therapeutic drug monitoring

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270 Phylogenetic Analysis of Klebsiella Species from Clinical Specimens from Nelson Mandela Academic Hospital in Mthatha, South Africa

Authors: Sandeep Vasaikar, Lary Obi

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Rapid and discriminative genotyping methods are useful for determining the clonality of the isolates in nosocomial or household outbreaks. Multilocus sequence typing (MLST) is a nucleotide sequence-based approach for characterising bacterial isolates. The genetic diversity and the clinical relevance of the drug-resistant Klebsiella isolates from Mthatha are largely unknown. For this reason, prospective, experimental study of the molecular epidemiology of Klebsiella isolates from patients being treated in Mthatha over a three-year period was analysed. Methodology: PCR amplification and sequencing of the drug-resistance-associated genes, and multilocus sequence typing (MLST) using 7 housekeeping genes mdh, pgi, infB, FusAR, phoE, gapA and rpoB were conducted. A total of 32 isolates were analysed. Results: The percentages of multidrug-resistant (MDR), extensively drug-resistance (XDR) and pandrug-resistant (PDR) isolates were; MDR 65.6 % (21) and XDR and PDR with 0 % each. In this study, K. pneumoniae was 19/32 (59.4 %). MLST results showed 22 sequence types (STs) were identified, which were further separated by Maximum Parsimony into 10 clonal complexes and 12 singletons. The most dominant group was Klebsiella pneumoniae with 23/32 (71.8 %) isolates, Klebsiella oxytoca as a second group with 2/32 (6.25 %) isolates, and a single (3.1 %) K. varricola as a third group while 6 isolates were of unknown sequences. Conclusions/significance: A phylogenetic analysis of the concatenated sequences of the 7 housekeeping genes showed that strains of K. pneumoniae form a distinct lineage within the genus Klebsiella, with K. oxytoca and K. varricola its nearest phylogenetic neighbours. With the analysis of 7 genes were determined 1 K. variicola, which was mistakenly identified as K. pneumoniae by phenotypic methods. Two misidentifications of K. oxytoca were found when phenotypic methods were used. No significant differences were observed between ESBL blaCTX-M, blaTEM and blaSHV groups in the distribution of Sequence types (STs) or Clonal complexes (CCs).

Keywords: phylogenetic analysis, phylogeny, klebsiella phylogenetic, klebsiella

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269 Petrologic and Geochemical Characteristics of Marine Sand Strip in the Proterozoic Chuanlinggou Formation of the North China

Authors: Yue Feng, Chun-jiang Wang, Zhi-long Huang

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The study of the sedimentary environment of Mesoproterozoic marine deposits in North China has attracted special attention in recent years. It is not clear that the sedimentary environment and the cause of formation of the sandstone strip and its internal carbonate cements and pyrite in the Mesoproterozoic Chuanlinggou Formation in North China. In this study, drilling core samples in North China were identified by microscopy, and their petrological characteristics such as mineral composition and structure were identified. The geochemical data of carbon and oxygen isotopes, total organic carbon (TOC) contents and total sulfur (TS) contents were obtained by processing and analyzing the samples. The samples are mainly quartz particles with low compositional maturity, combined with low value of TOC, it shows that the sedimentary environment of the sandy clastic is a sandy littoral sedimentary environment with relative strong hydrodynamic force, and then the sandstone strip in black shale are formed by the deposition of gravity flow. Analysis of TS values reflect sandstone bands formed in hypoxic environments. The carbonate cements and the pyrite in the sandstone belt are authigenic. The carbon isotope values of authigenic carbonate cements are negatively biased in comparison with the carbonate isotope of carbonate rocks in the same period, but it is more biased than the carbon isotopic values of anaerobic oxidation of methane (AOM) genetic carbonate rocks. Authigenic pyrite may be mainly due to the formation of HS- by the action of bacterial sulfate reduction (BSR) and Fe²⁺, their causes are in contact. This indicates that authigenic carbonate cements are mainly carbonate precipitates formed but are significantly affected by the effects of AOM. Summary, the sedimentary environment of the sandstone zone in the Chuanlinggou Formation in the North China is a shallow sea facies with iron rich and anoxic.

Keywords: sandstone strip, sedimentary environment, authigenic carbonate cements, authigenic pyrite, The Chuanlinggou group, North China

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268 Development of a Passive Solar Tomato Dryer with Movable Heat Storage System

Authors: Jacob T. Liberty, Wilfred I. Okonkwo

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The present study designed and constructed a post-harvest passive solar tomato dryer of dimension 176 x 152 x 54cm for drying tomato. Quality of the dried crop was evaluated and compared with the fresh ones. The solar dryer consist of solar collector (air heater), 110 x 61 x 10 x 10cm, the drying chamber, 102 x54cm, removal heat storage unit, 40 x 35 x 13cm and drying trays, 43 x 42cm. The physicochemical properties of this crop were evaluated before and after drying. Physicochemical properties evaluated includes moisture, protein, fat, fibre, ash, carbohydrate and vitamin C, contents. The fresh, open and solar dried samples were analysed for their proximate composition using the recommended method of AOAC. Also, statistical analysis of the data was conducted using analysis of variance (ANOVA) using completely Randomize Design (CRD) and means were separated by Duncan’s New Multiple Range test (DNMRT). Proximate analysis showed that solar dried tomato had significantly (P < 0.05) higher protein, fibre, ash, carbohydrate and vitamin C except for the fat content that was significantly (P < 0.05) higher for all the open sun dried samples than the solar dried and fresh product. The nutrient which is highly affected by sun drying is vitamin C. Result indicates that moisture loss in solar dried tomato was faster and lower than the open dried samples and as such makes the solar dried products of lesser tendency to mould and bacterial growth. Also, the open sun dried samples had to be carried into the sheltered place each time it rained. The solar dried produce is of high quality. Further processing of the dried crops will involve packaging for commercial purposes. This will also help in making these agricultural product available in a relatively cheap price in off season and also avert micronutrient deficiencies in diet especially among the low-income groups in Nigeria.

Keywords: tomato, passive solar dryer, physicochemical properties, removal heat storage

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267 Viability and Sensitivity of SFN6B (Host-Specific Bacteriophage) towards Shigella Flexneri in Various Water Samples

Authors: Siewchuiang Sia, Gimcheong Tan

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Bacteriophages are the most abundant and genetically diverse living entities on earth; they help in regulating and maintaining microbial diversity and balance in its natural ecosystem. In this study, the infectivity of SFN6B tailed phage was investigated in various water samples. Host bacteria (Shigella flexneri) were spiked in sterilized environmental and domestic water samples, followed by SFN6B treatment. Two incubation conditions were selected for this study, 37 oC and room temperature. S. flexneri and SFN6B viability were monitored hourly for consecutive 7 hours and extended viability study for consecutive 4 days. Absorbance of all bacteria spiked water samples were taken to monitor the bacteria count. Results showed reduction in the absorbance of the SFN6B treated water sample as compared to negative control, indicating reduction in bacterial count either due to negative growth or lysis by the lytic bacteriophage. Consistent with the result, SFN6B titer increases for first two days. However, prolong incubation of these cultures reaches equilibrium, between phage and bacteria. Temperature and water sample source also influence the interaction between S. flexneri and SFN6B. Stronger interaction was observed in 37oC as compared to room temperature, where higher bacteria count and phage titer increase were recorded. Availability of nutrient in water sample also plays a crucial role in the interaction between bacteria and phage. Higher nutrient level, such as lake and river waters were observed to give better infectivity and viability of both bacteria and phage as compared to tab water. It is believed that S. flexneri continue to remain viable and able to grow in the present of SFN6B bacteriophage, but the number was closely regulated by surrounding phages. This allows better understanding of the characteristics of SFN6B that could serve as the basis for future studies and applications.

Keywords: bacteriophage, Shigella flexneri, infection, microbial diversity

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266 Paper-Like and Battery Free Sensor Patches for Wound Monitoring

Authors: Xiaodi Su, Xin Ting Zheng, Laura Sutarlie, Nur Asinah binte Mohamed Salleh, Yong Yu

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Wound healing is a dynamic process with multiple phases. Rapid profiling and quantitative characterization of inflammation and infection remain challenging. We have developed paper-like battery-free multiplexed sensors for holistic wound assessment via quantitative detection of multiple inflammation and infection markers. In one of the designs, the sensor patch consists of a wax-printed paper panel with five colorimetric sensor channels arranged in a pattern resembling a five-petaled flower (denoted as a ‘Petal’ sensor). The five sensors are for temperature, pH, trimethylamine, uric acid, and moisture. The sensor patch is sandwiched between a top transparent silicone layer and a bottom adhesive wound contact layer. In the second design, a palm-like-shaped paper strip is fabricated by a paper-cutter printer (denoted as ‘Palm’ sensor). This sensor strip carries five sensor regions connected by a stem sampling entrance that enables rapid colorimetric detection of multiple bacteria metabolites (aldehyde, lactate, moisture, trimethylamine, tryptophan) from wound exudate. For both the “\’ Petal’ and ‘Palm’ sensors, color images can be captured by a mobile phone. According to the color changes, one can quantify the concentration of the biomarkers and then determine wound healing status and identify/quantify bacterial species in infected wounds. The ‘Petal’ and ‘Palm’ sensors are validated with in-situ animal and ex-situ skin wound models, respectively. These sensors have the potential for integration with wound dressing to allow early warning of adverse events without frequent removal of the plasters. Such in-situ and early detection of non-healing condition can trigger immediate clinical intervention to facilitate wound care management.

Keywords: wound infection, colorimetric sensor, paper fluidic sensor, wound care

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265 The Neuropsychology of Obsessive Compulsion Disorder

Authors: Mia Bahar, Özlem Bozkurt

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Obsessive-compulsive disorder (OCD) is a typical, persistent, and long-lasting mental health condition in which a person experiences uncontrollable, recurrent thoughts (or "obsessions") and/or activities (or "compulsions") that they feel compelled to engage in repeatedly. Obsessive-compulsive disorder is both underdiagnosed and undertreated. It frequently manifests in a variety of medical settings and is persistent, expensive, and burdensome. Obsessive-compulsive neurosis was long believed to be a condition that offered valuable insight into the inner workings of the unconscious mind. Obsessive-compulsive disorder is now recognized as a prime example of a neuropsychiatric condition susceptible to particular pharmacotherapeutic and psychotherapy therapies and mediated by pathology in particular neural circuits. An obsessive-compulsive disorder which is called OCD, usually has two components, one cognitive and the other behavioral, although either can occur alone. Obsessions are often repetitive and intrusive thoughts that invade consciousness. These obsessions are incredibly hard to control or dismiss. People who have OCD often engage in rituals to reduce anxiety associated with intrusive thoughts. Once the ritual is formed, the person may feel extreme relief and be free from anxiety until the thoughts of contamination intrude once again. These thoughts are strengthened through a manifestation of negative reinforcement because they allow the person to avoid anxiety and obscurity. These thoughts are described as autogenous, meaning they most likely come from nowhere. These unwelcome thoughts are related to actions which we can describe as Thought Action Fusion. The thought becomes equated with an action, such as if they refuse to perform the ritual, something bad might happen, and so people perform the ritual to escape the intrusive thought. In almost all cases of OCD, the person's life gets extremely disturbed by compulsions and obsessions. Studies show OCD is an estimated 1.1% prevalence, making it a challenging issue with high co-morbidities with other issues like depressive episodes, panic disorders, and specific phobias. The first to reveal brain anomalies in OCD were numerous CT investigations, although the results were inconsistent. A few studies have focused on the orbitofrontal cortex (OFC), anterior cingulate gyrus (AC), and thalamus, structures also implicated in the pathophysiology of OCD by functional neuroimaging studies, but few have found consistent results. However, some studies have found abnormalities in the basal ganglion. There have also been some discussions that OCD might be genetic. OCD has been linked to families in studies of family aggregation, and findings from twin studies show that this relationship is somewhat influenced by genetic variables. Some Research has shown that OCD is a heritable, polygenic condition that can result from de novo harmful mutations as well as common and unusual variants. Numerous studies have also presented solid evidence in favor of a significant additive genetic component to OCD risk, with distinct OCD symptom dimensions showing both common and individual genetic risks.

Keywords: compulsions, obsessions, neuropsychiatric, genetic

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264 Ectoine: A Compatible Solute in Radio-Halophilic Stenotrophomonas sp. WMA-LM19 Strain to Prevent Ultraviolet-Induced Protein Damage

Authors: Wasim Sajjad, Manzoor Ahmad, Sundas Qadir, Muhammad Rafiq, Fariha Hasan, Richard Tehan, Kerry L. McPhail, Aamer Ali Shah

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Aim: This study aims to investigate the possible radiation protective role of a compatible solute in the tolerance of radio-halophilic bacterium against stresses, like desiccation and exposure to ionizing radiation. Methods and Results: Nine different radio-resistant bacteria were isolated from desert soil, where strain WMA-LM19 was chosen for detailed studies on the basis of its high tolerance for ultraviolet radiation among all these isolates. 16S rRNA gene sequencing indicated that the bacterium was closely related to Stenotrophomonas sp. (KT008383). A bacterial milking strategy was applied for extraction of intracellular compatible solutes in 70% (v/v) ethanol, which were purified by high-performance liquid chromatography (HPLC). The compound was characterized as ectoine by 1H and 13C nuclear magnetic resonance (NMR), and mass spectrometry (MS). Ectoine demonstrated more efficient preventive activity (54.80%) to erythrocyte membranes and also inhibited oxidative damage to proteins and lipids in comparison to the standard ascorbic acid. Furthermore, a high level of ectoine-mediated protection of bovine serum albumin against ionizing radiation (1500-2000 Jm-2) was observed, as indicated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Conclusion: The results indicated that ectoine can be used as a potential mitigator and radio-protective agent to overcome radiation- and salinity-mediated oxidative damage in extreme environments. Significance and Impact of the Study: This study shows that ectoine from radio-halophiles can be used as a potential source in topical creams as sunscreen. The investigation of ectoine as UV protectant also changes the prospective that radiation resistance is specific only to molecular adaptation.

Keywords: ectoine, anti-oxidant, stenotrophomonas sp., ultraviolet radiation

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263 Antimicrobial Effect of Toothpastes Containing Fluoride, Xylitol or Xylitol-Probiotic on Salivary Streptococcus mutans and Lactobacillus in Children

Authors: Eda Arat Maden, Ceyhan Altun, Bilal Ozmen, Feridun Basak

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Purpose: The purpose of this study was to compare the antimicrobial effect of toothpastes containing fluoride, xylitol or xylitol-probiotic in vivo, Streptococcus mutans and Lactobacillus in 13-15 years old children. Method: The study consisted of 60 pediatric patients were randomly divided into 3 groups of 20 each. Group 1 received fluoride toothpaste (Colgate Max Fresh), group 2 used xylitol toothpaste (Xyliwhite) and group 3 used xylitol-probiotic toothpaste (PerioBiotic). Subjects were asked to use the allocated dentifrice two times a day, for 6 weeks. We performed tests on the samples of saliva in the beginning of the study and after 6 weeks’ duration following the use of toothpaste. Result and Conclusion: All of the participants of the study stated that they brushed their teeth well twice a day by using the toothpastes given to them for 6 weeks. Majority of the subjects had high counts of salivary mutans streptococci and Lactobacillus at baseline. When the number of cariogenic bacteria (S. mutans and Lactobacillus) at the start of the PerioBiotic Probiotic toothpaste usage are compared with the results measured after 6 weeks, an important decrease is observed in the S. mutans and Lactobacillus bacteria according to the CRT Tests. After the 6-week use of Probiotic toothpaste, the S. mutans (≥105) decreased to 20% from 75% in the group with S. mutans and Lactobacillus (≥105) decreased to 30% from 60% in the group with Lactobacillus. In addition, an important decrease was recorded in the participants with the S. mutans percentage (80% - 45%) and Lactobacillus (70% - 55%) after using the Colgate Max Fresh toothpaste for six weeks. On the other hand, it was determined with the Chi-square that there were not important changes between the Xyliwhite toothpaste group and the other groups with S. mutans (80% - 75%) and Lactobacillus (75% -65%). It was also determined after the comparison of the groups that the decrease in the S. mutans was higher than the group using PerioBiotic Probiotic toothpaste at a significant level, when compared with the Colgate Max Fresh toothpaste and Xyliwhite toothpaste. S. mutans were more sensitive to the antimicrobial activity of the PerioBiotic Probiotic toothpaste and to the Colgate Max Fresh toothpaste when compared with the Lactobacillus. In the light of the data obtained in this in vivo study, the use of probiotics ensure the balance between the bacterial flora in the oral cavity.

Keywords: lactobacillus, probiotic, Streptococcus mutans, toothpaste

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262 In vitro Evaluation of Prebiotic Potential of Wheat Germ

Authors: Lígia Pimentel, Miguel Pereira, Manuela Pintado

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Wheat germ is a by-product of wheat flour refining. Despite this by-product being a source of proteins, lipids, fibres and complex carbohydrates, and consequently a valuable ingredient to be used in Food Industry, only few applications have been studied. The main goal of this study was to assess the potential prebiotic effect of natural wheat germ. The prebiotic potential was evaluated by in vitro assays with individual microbial strains (Lactobacillus paracasei L26 and Lactobacillus casei L431). A simulated model of the gastrointestinal digestion was also used including the conditions present in the mouth (artificial saliva), oesophagus–stomach (artificial gastric juice), duodenum (artificial intestinal juice) and ileum. The effect of natural wheat germ and wheat germ after digestion on the growth of lactic acid bacteria was studied by growing those microorganisms in de Man, Rogosa and Sharpe (MRS) broth (with 2% wheat germ and 1% wheat germ after digestion) and incubating at 37 ºC for 48 h with stirring. A negative control consisting of MRS broth without glucose was used and the substrate was also compared to a commercial prebiotic fructooligosaccharides (FOS). Samples were taken at 0, 3, 6, 9, 12, 24 and 48 h for bacterial cell counts (CFU/mL) and pH measurement. Results obtained showed that wheat germ has a stimulatory effect on the bacteria tested, presenting similar (or even higher) results to FOS, when comparing to the culture medium without glucose. This was demonstrated by the viable cell counts and also by the decrease on the medium pH. Both L. paracasei L26 and L. casei L431 could use these compounds as a substitute for glucose with an enhancement of growth. In conclusion, we have shown that wheat germ stimulate the growth of probiotic lactic acid bacteria. In order to understand if the composition of gut bacteria is altered and if wheat germ could be used as potential prebiotic, further studies including faecal fermentations should be carried out. Nevertheless, wheat germ seems to have potential to be a valuable compound to be used in Food Industry, mainly in the Bakery Industry.

Keywords: by-products, functional ingredients, prebiotic potential, wheat germ

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261 Biological Activities of Protease Inhibitors from Cajanus cajan and Phaseolus limensis

Authors: Tooba N. Shamsi, Romana Perveen, Sadaf Fatima

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Protease Inhibitors (PIs) are widespread in nature, produced by animals, plants and microorganisms. They play vital role in various biological activities by keeping a check on activity of proteases. Present study aims to investigate antioxidant and anti-inflammatory properties of PPI from Cajanus cajan (CCTI) and Phaseolus limensis (LBTI). PPI was purified from C. cajan (PUSA-992) by ammonium sulfate precipitation followed by ion exchange chromatography. The anti-oxidant activity was analyzed by two most common radical scavenging assays of FRAP (ferric reducing antioxidant power) and DPPH (1,1- diphenyl-2-picrylhydrazyl). Also, in-vitro anti-inflammatory activity was evaluated using albumin denaturation assay and membrane stabilization assay at different concentrations. Ascorbic acid and aspirin were used as a standards for antioxidant and anti-inflammatory assays respectively. The PPIs were also checked for antimicrobial activity against a number of bacterial strains. The CCTI and LBTI showed DPPH radical scavenging activity in a concentration–dependent manner with IC50 values 544 µg/ml and 506 µg/ml respectively comparative to ascorbic acid which was 258 µg/ml. Following FRAP assay, it was evaluated that LBTI had 87.5% and CCTI showed 84.4% antioxidant activity, taking value of standard ascorbic acid to be 100%. The PPIs also showed in-vitro anti‐inflammatory activity by inhibiting the heat induced albumin denaturation with IC50 values of 686 µg/ml and 615 µg/ml for CCTI and LBTI respectively compared to the standard (aspirin) which was 70.8 µg/ml. Red blood cells membrane stabilization with IC50 values of 641 µg/ml and 587 µg/ml for CCTI and LBTI respectively against aspirin which showed IC50 value of 70.4 µg/ml. PPIs showed antibacterial activity against 7 known strains while there was apparently no action against fungi.

Keywords: Cajanus cajan, Phaseolus limensis, Lima beans, protein protease inhibitor, antioxidant, anti-inflammatory, antimicrobial activity

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260 Biochemical Characterization of CTX-M-15 from Enterobacter cloacae and Designing a Novel Non-β-Lactam-β-Lactamase Inhibitor

Authors: Mohammad Faheem, M. Tabish Rehman, Mohd Danishuddin, Asad U. Khan

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The worldwide dissemination of CTX-M type β-lactamases is a threat to human health. Previously, we have reported the spread of blaCTX-M-15 gene in different clinical strains of Enterobacteriaceae from the hospital settings of Aligarh in north India. In view of the varying resistance pattern against cephalosporins and other β-lactam antibiotics, we intended to understand the correlation between MICs and catalytic activity of CTX-M-15. In this study, steady-state kinetic parameters and MICs were determined on E. coli DH5α transformed with blaCTX-M-15 gene that was cloned from Enterobacter cloacae (EC-15) strain of clinical background. The effect of conventional β-lactamase inhibitors (clavulanic acid, sulbactam and tazobactam) on CTX-M-15 was also studied. We have found that tazobactam is the best among these inhibitors against CTX-M-15. The inhibition characteristic of tazobactam is defined by its very low IC50 value (6 nM), high affinity (Ki = 0.017 µM) and better acylation efficiency (k+2/K9 = 0.44 µM-1s-1). It forms an acyl-enzyme covalent complex, which is quite stable (k+3 = 0.0057 s-1). Since increasing resistance has been reported against conventional b-lactam antibiotic-inhibitor combinations, we aspire to design a non-b-lactam core containing b-lactamase inhibitor. For this, we screened ZINC database and performed molecular docking to identify a potential non-β-lactam based inhibitor (ZINC03787097). The MICs of cephalosporin antibiotics in combination with this inhibitor gave promising results. Steady-state kinetics and molecular docking studies showed that ZINC03787097 is a reversible inhibitor which binds non-covalently to the active site of the enzyme through hydrogen bonds and hydrophobic interactions. Though, it’s IC50 (180 nM) is much higher than tazobactam, it has good affinity for CTX-M-15 (Ki = 0.388 µM). This study concludes that ZINC03787097 compound can be used as seed molecule to design more efficient non-b-lactam containing b-lactamase inhibitor that could evade pre-existing bacterial resistance mechanisms.

Keywords: ESBL, non-b-lactam-b-lactamase inhibitor, bioinformatics, biomedicine

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259 Case Report: Mandibular Area Abscesses in Calves

Authors: Dovilė Bačėninaitė, Karina Džermeikaitė, Justinas Kirvela, Ramūnas Antanaitis

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Bacteria are often present in the mouth of cattle. Some of them can cause abscesses. Starting with severe swelling of the mouth, muscle spasm, or locked jaw, it can lead to inability to open its mouth, move the neck, cause pain while eating. While the calf is unable to eat properly, it becomes more susceptible to infectious diseases, lower weight gain can be observed. Abscesses can be considered as a continuum of oral disease, whereby early stages of the lumpy jaw could proceed from gingivitis to periodontal disease. In the event of tissue damage, bacteria can enter the bloodstream, even cause sepsis. The most common lesions occur when animals eat sharp grass, coarse fodder, sharp, piercing foreign bodies (this is especially common for calves when they are trying to eat inedible objects). A crossbred Holstein calf presented with a history of proliferative outgrowth in the mandibular region. On clinical examination, needle aspiration, mandibular swelling revealed sticky, white curd-like fluid containing. Pus bacteriology revealed gram-negative cocci. They were sensitive to amoxicillin, cephalexin, enrofloxacin, ceftiofur. Blood morphology was in physiological ranges. The calf was treated surgically. The growth was excised, the puss drained and the wound was flushed with potassium permanganate solution (0,01%). A week after clinical surgery examination was performed. The swelling was decreased. Superficial bacterial infections are often associated with poor hygiene, which should be improved before treatment is commenced. Clipping away dirty hair and gently washing affected areas of skin daily with solutions such as povidone-iodine, potassium permanganate is effective. Appropriate antibiotic therapy, based on sensitivity testing, may be used where there is evidence of systemic illness.

Keywords: calf, abscess, lumpy jaw, pus, Streptococcus, Staphylococcus, Actinobacillus, infection

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258 Investigation of Enterotoxigenic Staphylococcus aureus in Kitchen of Catering

Authors: Çiğdem Sezer, Aksem Aksoy, Leyla Vatansever

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This study has been done for the purpose of evaluation of public health and identifying of enterotoxigenic Staphyloccocus aureus in kitchen of catering. In the kitchen of catering, samples have been taken by swabs from surface of equipments which are in the salad section, meat section and bakery section. Samples have been investigated with classical cultural methods in terms of Staphyloccocus aureus. Therefore, as a 10x10 cm area was identified (salad, cutting and chopping surfaces, knives, meat grinder, meat chopping surface) samples have been taken with sterile swabs with helping FTS from this area. In total, 50 samples were obtained. In aseptic conditions, Baird-Parker agar (with egg yolk tellurite) surface was seeded with swabs. After 24-48 hours of incubation at 37°C, the black colonies with 1-1.5 mm diameter and which are surrounded by a zone indicating lecithinase activity were identified as S. aureus after applying Gram staining, catalase, coagulase, glucose and mannitol fermentation and termonuclease tests. Genotypic characterization (Staphylococcus genus and S.aureus species spesific) of isolates was performed by PCR. The ELISA test was applied to the isolates for the identification of staphylococcal enterotoxins (SET) A, B, C, D, E in bacterial cultures. Measurements were taken at 450 nm in an ELISA reader using an Ridascreen-Total set ELISA test kit (r-biopharm R4105-Enterotoxin A, B, C, D, E). The results were calculated according to the manufacturer’s instructions. A total of 50 samples of 97 S. aureus was isolated. This number has been identified as 60 with PCR analysis. According to ELISA test, only 1 of 60 isolates were found to be enterotoxigenic. Enterotoxigenic strains were identified from the surface of salad chopping and cutting. In the kitchen of catering, S. aureus identification indicates a significant source of contamination. Especially, in raw consumed salad preparation phase of contamination is very important. This food can be a potential source of food-borne poisoning their terms, and they pose a significant risk to consumers have been identified.

Keywords: Staphylococcus aureus, enterotoxin, catering, kitchen, health

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257 Phenomena-Based Approach for Automated Generation of Process Options and Process Models

Authors: Parminder Kaur Heer, Alexei Lapkin

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Due to global challenges of increased competition and demand for more sustainable products/processes, there is a rising pressure on the industry to develop innovative processes. Through Process Intensification (PI) the existing and new processes may be able to attain higher efficiency. However, very few PI options are generally considered. This is because processes are typically analysed at a unit operation level, thus limiting the search space for potential process options. PI performed at more detailed levels of a process can increase the size of the search space. The different levels at which PI can be achieved is unit operations, functional and phenomena level. Physical/chemical phenomena form the lowest level of aggregation and thus, are expected to give the highest impact because all the intensification options can be described by their enhancement. The objective of the current work is thus, generation of numerous process alternatives based on phenomena, and development of their corresponding computer aided models. The methodology comprises: a) automated generation of process options, and b) automated generation of process models. The process under investigation is disintegrated into functions viz. reaction, separation etc., and these functions are further broken down into the phenomena required to perform them. E.g., separation may be performed via vapour-liquid or liquid-liquid equilibrium. A list of phenomena for the process is formed and new phenomena, which can overcome the difficulties/drawbacks of the current process or can enhance the effectiveness of the process, are added to the list. For instance, catalyst separation issue can be handled by using solid catalysts; the corresponding phenomena are identified and added. The phenomena are then combined to generate all possible combinations. However, not all combinations make sense and, hence, screening is carried out to discard the combinations that are meaningless. For example, phase change phenomena need the co-presence of the energy transfer phenomena. Feasible combinations of phenomena are then assigned to the functions they execute. A combination may accomplish a single or multiple functions, i.e. it might perform reaction or reaction with separation. The combinations are then allotted to the functions needed for the process. This creates a series of options for carrying out each function. Combination of these options for different functions in the process leads to the generation of superstructure of process options. These process options, which are formed by a list of phenomena for each function, are passed to the model generation algorithm in the form of binaries (1, 0). The algorithm gathers the active phenomena and couples them to generate the model. A series of models is generated for the functions, which are combined to get the process model. The most promising process options are then chosen subjected to a performance criterion, for example purity of product, or via a multi-objective Pareto optimisation. The methodology was applied to a two-step process and the best route was determined based on the higher product yield. The current methodology can identify, produce and evaluate process intensification options from which the optimal process can be determined. It can be applied to any chemical/biochemical process because of its generic nature.

Keywords: Phenomena, Process intensification, Process models , Process options

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256 Impact of a Structured Antimicrobial Stewardship Program in a North-East Italian Hospital

Authors: Antonio Marco Miotti, Antonella Ruffatto, Giampaola Basso, Antonio Madia, Giulia Zavatta, Emanuela Salvatico, Emanuela Zilli

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A National Action Plan to fight antimicrobial resistance was launched in Italy in 2017. In order to reduce inappropriate exposure to antibiotics and infections from multi-drug resistant bacteria, it is essential to set up a structured system of surveillance and monitoring of the implementation of National Action Plan standards, including antimicrobial consumption, with a special focus on quinolones, third generation cephalosporins and carbapenems. A quantitative estimate of antibiotic consumption (defined daily dose - DDD - consumption per 100 days of hospitalization) has been provided by the Pharmaceutical Service to the Hospital of Cittadella, ULSS 6 Euganea – Health Trust (District of Padua) for the years 2019 (before the pandemic), 2020 and 2021 for all classes of antibiotics. Multidisciplinary meetings have been organized monthly by the local Antimicrobial Stewardship Group. Between 2019 and 2021, an increase in the consumption of carbapenems in the Intensive Care Unit (from 12.2 to 18.2 DDD, + 49.2%) and a decrease in Medical wards (from 5.3 to 2.6 DDD, - 50.9%) was reported; a decrease in the consumption of quinolones in Intensive Care Unit (from 17.2 to 10.8 DDD, - 37.2%), Medical wards (from 10.5 to 6.6 DDD, - 37.1%) and Surgical wards (from 10.2 to 9.3 DDD, - 8.8%) was highlighted; an increase in the consumption of third generation cephalosporins in Medical wards (from 18.1 to 22.6 DDD, + 24,1%) was reported. Finally, after an increase in the consumption of macrolides between 2020 and 2019, in 2021, a decrease was reported in the Intensive Care Unit (DDD: 8.0 in 2019, 18.0 in 2020, 6.4 in 2021) and Medical wards (DDD: 9.0 in 2019, 13.7 in 2020, 10.9 in 2021). Constant monitoring of antimicrobial consumption and timely identifying of warning situations that may need a specific intervention are the cornerstone of Antimicrobial Stewardship programs, together with analysing data on bacterial resistance rates and infections from multi-drug resistant bacteria.

Keywords: carbapenems, quinolones, antimicrobial, stewardship

Procedia PDF Downloads 146
255 Satureja bachtiarica Bunge Induce Apoptosis via Mitochondrial Intrinsic Pathway and G1 Cell Cycle Arrest

Authors: Hamed Karimian, Noraziah Nordin, Mohamad Ibrahim Noordin, Syam Mohan, Mahboubeh Razavi, Najihah Mohd Hashim, Happipah Mohd Ali

Abstract:

Satureja bachtiarica Bunge is a perennial medicinal plant belonging to the Lamiaceae family and endemic species in Iran. Satureja bachtiarica Bunge with the local name of Marzeh koohi is edible vegetable use as flavoring agent, anti-bacterial and to relieve cough and indigestion. In this study, the anti-cancer effect of Satureja bachtiarica Bunge on the MDA-MB-231 cell line as an Breast cancer cell model has been analyzed for the first time. Satureja bachtiarica Bunge was extracted using different solvents in the order of increasing polarity. Cytotoxicity activity of hexane extract of Satureja bachtiarica Bunge (SBHE) was observed using MTT assay. Acridine orange/Propidium iodide staining was used to detect early apoptosis; Annexin-V-FITC assay was carried out to observe the detection of cell-surface Phosphatidylserine (PS), with Annexin-Vserving as a marker for apoptotic cells. Caspase 3/7, 8 and-9 assays showed significantly activation of caspase-9 where lead intrinsic mitochondrial pathway. Bcl-2/Bax expressions and cell cycle arrest were also investigated. SBHE had exhibited significantly higher cytotoxicity against MDA-MB-231 Cell line compare to other cell lines. A significant increase in chromatin condensation in the cell nucleus was observed by fluorescence analysis. Treatment of MDA-MB-231 cells with SBHE encouraged apoptosis, by down-regulating Bcl-2 and up-regulating Bax, which lead the activation of caspase 9. Moreover, SBHE treatment significantly arrested MDA-MB-231 cells in the G1 phase. Together, the results presented in this study demonstrated that SBHE inhibited the proliferation of MDA-MB-231 cells, leading cell cycle arrest and programmed cell death, which was confirmed to be through the mitochondrial pathway.

Keywords: Satureja bachtiarica Bunge, MDA-MB-231, apoptosis, annexin-V, cell cycle

Procedia PDF Downloads 332
254 Selective Recovery and Molecular Identification of Laccase-Producing Bacteria from Selected Terrestrial and Aquatic Milieu in the Eastern Cape, South Africa: Toward the Production of Environmentally Relevant Biocatalysts

Authors: John Onolame Unuofin, Uchechukuw U. Nwodo, Anthony I. Okoh

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Laccase is constantly gaining status as important biocatalyst in biotechnology. The illimitable potential of its industrial applications and the corresponding aggressive need for phenomenal volumes of extracellularly secreted laccases have called for its interminable production from sources which are able to meet this demand within a relatively short period of time, preferably bacteria. In response to this call, this study was designed to source for laccase-producing bacteria from different environmental matrices. Three sampling environments were chosen such as wastewater treatment plants, University of Fort Hare vicinity and the Hogback woodland, all within the Eastern Cape, South Africa. Samples such as effluents, sediments, leaf litters, degrading wood and rock scrapings were selectively enriched with some model aromatic compounds and were further screened qualitatively and quantitatively on five phenolic substrates ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), Guaiacol, 1-Naphthol, Potassium Ferric Cyanide and Syringaldazine). Basis for selection was their ability to elicit a colour change on at least three of the above mentioned agar based assay substrates. The choice isolates were further identified based on 16S rRNA molecular identification techniques. 33 isolates were screened out of the 40 representative distinct colonies during the qualitative plate screens, while quantitative screens selected out 11 bacterial isolates. They were, based on molecular identification, desginated as members of the genera Pseudomonas, Stenotrophomonas and Citrobacter of the gammaproteobacteria and Bordetalla and Achromobacter of the betaproteobacteria respectively. We therefore conclude based on our outcomes that we may have isolated efficient laccase-producing bacteria, which might be of beneficial significance in catalysis and biotechnology.

Keywords: beta proteobacteria, catalysis, gammaproteobacteria, laccase

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253 ATR-IR Study of the Mechanism of Aluminum Chloride Induced Alzheimer Disease - Curative and Protective Effect of Lepidium sativum Water Extract on Hippocampus Rats Brain Tissue

Authors: Maha J. Balgoon, Gehan A. Raouf, Safaa Y. Qusti, Soad S. Ali

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The main cause of Alzheimer disease (AD) was believed to be mainly due to the accumulation of free radicals owing to oxidative stress (OS) in brain tissue. The mechanism of the neurotoxicity of Aluminum chloride (AlCl3) induced AD in hippocampus Albino wister rat brain tissue, the curative & the protective effects of Lipidium sativum group (LS) water extract were assessed after 8 weeks by attenuated total reflection spectroscopy ATR-IR and histologically by light microscope. ATR-IR results revealed that the membrane phospholipid undergo free radical attacks, mediated by AlCl3, primary affects the polyunsaturated fatty acids indicated by the increased of the olefinic -C=CH sub-band area around 3012 cm-1 from the curve fitting analysis. The narrowing in the half band width(HBW) of the sνCH2 sub-band around 2852 cm-1 due to Al intoxication indicates the presence of trans form fatty acids rather than gauch rotomer. The degradation of hydrocarbon chain to shorter chain length, increasing in membrane fluidity, disorder and decreasing in lipid polarity in AlCl3 group were indicated by the detected changes in certain calculated area ratios compared to the control. Administration of LS was greatly improved these parameters compared to the AlCl3 group. Al influences the Aβ aggregation and plaque formation, which in turn interferes to and disrupts the membrane structure. The results also showed a marked increase in the β-parallel and antiparallel structure, that characterize the Aβ formation in Al-induced AD hippocampal brain tissue, indicated by the detected increase in both amide I sub-bands around 1674, 1692 cm-1. This drastic increase in Aβ formation was greatly reduced in the curative and protective groups compared to the AlCl3 group and approaches nearly the control values. These results were supported too by the light microscope. AlCl3 group showed significant marked degenerative changes in hippocampal neurons. Most cells appeared small, shrieked and deformed. Interestingly, the administration of LS in curative and protective groups markedly decreases the amount of degenerated cells compared to the non-treated group. Also the intensity of congo red stained cells was decreased. Hippocampal neurons looked more/or less similar to those of control. This study showed a promising therapeutic effect of Lipidium sativum group (LS) on AD rat model that seriously overcome the signs of oxidative stress on membrane lipid and restore the protein misfolding.

Keywords: aluminum chloride, alzheimer disease, ATR-IR, Lipidium sativum

Procedia PDF Downloads 356
252 Effect of Media Osmolarity on Vi Biosynthesis on Salmonella enterica serovar Typhi Strain C6524 Cultured on Batch System

Authors: Dwi Arisandi Wijaya, Ernawati Arifin Giri-Rachman, Neni Nurainy

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Typhoid fever disease can be prevented by using a polysaccharide-based vaccine Vi which is a virulence factor of S.typhi. To produce high yield Vi polysaccharide from bacteria, it is important to know the biosynthesis of Vi polysaccharide and the regulators involved. In the In vivo condition, S. typhi faces different osmolarity, and the bacterial two-component system OmpR-EnvZ, regulate by up and down Capsular Vi polysaccharide biosynthesis. A high yielded Vi Polysaccharide strain, S. typhi strain C6524 used to study the effect of media osmolarity on Vi polysaccharide biosynthesis and the osmoregulation pattern of S. typhi strain C6524. The methods were performed by grown S. typhi strain C6524 grown on medium with 50 mM, 100 mM, and 150 mM osmolarity with the batch system. Vi polysaccharide concentration was measured by ELISA method. For further investigation of the osmoregulation pattern of strain C6524, the osmoregulator gene, OmpR, has been isolated and sequenced using the specific primer of the OmpR gene. Nucleotide sequence analysis is done with BLAST and Lallign. Amino Acid sequence analysis is done with Prosite and Multiple Sequence Alignment. The results of cultivation showed the average content of polysaccharide Vi for 50 mM, 100 mM, and 150 mM osmolarities 11.49 μg/mL, 12.06 μg/mL, and 14.53 μg/mL respectively. Analysis using Anova stated that the osmolarity treatment of 150 mM significantly affects Vi content. Analysis of nucleotide sequences shows 100% identity between S. typhi strain C6524 and Ty2. Analysis of amino acid sequences shows that the OmpR response regulator protein of the C6524 strain also has a α4-β5-α5 motif which is important for the regulatory activation system when phosphorylation occurs by domain kinase. This indicates that the regulator osmolarity response of S. typhi strain C6524 has no difference with the response regulator owned by S. typhi strain Ty2. A high Vi response rate in the 150 mM osmolarity treatment requires further research for RcsB-RcsC, another two-component system involved in Vi Biosynthesis.

Keywords: osmoregulator, OmpR, Salmonella, Vi polysaccharide

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251 Microbial Quality Assessment of Indian White Shrimp, Penaeus Indicus from Southwest Bangladesh

Authors: Saima Sharif Nilla, Mahmudur Rahman Khan, Anisur Rahman Khan, Ghulam Mustafa1

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The microbial quality of Indian white shrimp (Peneaus indicus) from Bagerhat, Khulna and Satkhira of southwest Bangladesh was assessed where the parameters varied with different sources and the quality was found to be poor for Satkhira shrimp samples. Shrimp samples in fresh condition were collected to perform the microbial assessment and 10 pathogenic isolates for antibiotic sensitivity test to 12 antibiotics. The results show that total bacterial count of all the samples were beyond the acceptable limit 105 cfu/g. In case of total coliform and E. coli density, no substantial difference (p<0.5) was found between the different shrimp samples from different districts and also high quantity of TC exceeding the limit (>102 cfu/g) proves the poor quality of shrimp. The FC abundance found in shrimps of Bagerhat and Satkhira was similar and significantly higher (p<0.5) than that of Khulna samples. No significant difference (p<0.5) was found among the high density of Salmonella-Shigella, Vibrio spp., and Staphylococcus spp. of the shrimp samples from the source places. In case of antibiotic sensitivity patterns, all of them were resistant to ampicillin, Penicillin and sensitive to kanamycin. Most of the isolates were frequently sensitive to ciprofloxacin and streptomycin in the sensitivity test. In case of nutritional composition, no significant difference (t-test, p<0.05) was found among protein, lipid, moisture and ash contents of shrimp samples. The findings prove that shrimp under this study was more or less contaminated and samples from Satkhira were highly privileged with food borne pathogens which confirmed the unhygienic condition of the shrimp farms as well as the presence of antibiotic resistance bacteria in shrimp fish supposed to threat food safety and deteriorate the export quality.

Keywords: food borne pathogens, satkhira, penaeus indicus, antibiotic sensitivity, southwest Bangladesh, food safety

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250 Theory of Negative Trigger: The Contract between Oral Probiotics and Immune System

Authors: Cliff Shunsheng Han

Abstract:

Identifying the direct allergy cause that can be easily mitigated is the foundation to stop the allergy epidemic that has been started in the seventies. It has confirmed that the personal and social hygiene practices are associated with the allergy prevalence. But direct causes have been found, and proposed translational measures have not been effective. This study, assisted by a particular case of allergies, has seen the direct cause of allergies, developed a valid test resulted in lasting relief for allergies, and constructed theory describing general relationship between microbiota and host immune system. Saliva samples were collected from a subject for three years during which time the person experienced yearlong allergy, seasonal allergy, and remission of allergy symptoms. Bacterial DNA was extracted and 16S rRNA genes were profiled with Illumina sequencing technology. The analyzing results indicate that the possible direct cause of allergy is the lacking probiotic bacteria in the oral cavity, such as genera Streptococcus and Veilonella, that can produce metabolites to pacify immune system. Targeted promotion of those bacteria with a compound designed for them, has led to lasting remissions of allergic rhinitis. During the development of the translational measure, the subject's oral biofilm was completely destructed by a moderate fever due to an unrelated respiratory infection. The incident not only facilitated the development of the heat based microbiota reseeding procedure but also indicated a possible natural switch that subsequently increases the efficacy of the immune system previously restrained by metabolites from microbiota. These results lead to the proposal of a Theory of Negative Trigger (TNT) to describe the relationship between oral probiotics and immune system, in which probiotics are the negative trigger that will release the power of immune system when removed by fever or modern lifestyles. This study could open doors leading to further understanding of how the immune system functions under the influence of microbiota as well as validate simple traditional practices for healthy living.

Keywords: oral microbiome, allergy, immune system, infection

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249 Greening the Blue: Enzymatic Degradation of Commercially Important Biopolymer Dextran Using Dextranase from Bacillus Licheniformis KIBGE-IB25

Authors: Rashida Rahmat Zohra, Afsheen Aman, Shah Ali Ul Qader

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Commercially important biopolymer, dextran, is enzymatically degraded into lower molecular weight fractions of vast industrial potential. Various organisms are associated with dextranase production, among which fungal, yeast and bacterial origins are used for commercial production. Dextranases are used to remove contaminating dextran in sugar processing industry and also used in oral care products for efficient removal of dental plaque. Among the hydrolytic products of dextran, isomaltooligosaccharides have prebiotic effect in humans and reduces the cariogenic effect of sucrose in oral cavity. Dextran derivatives produced by hydrolysis of high molecular polymer are also conjugated with other chemical and metallic compounds for usage in pharmaceutical, fine chemical industry, cosmetics, and food industry. Owing to the vast application of dextran and dextranases, current study focused on purification and analysis of kinetic parameters of dextranase from a newly isolated strain of Bacillus licheniformis KIBGE-IB25. Dextranase was purified up to 35.75 folds with specific activity of 1405 U/mg and molecular weight of 158 kDa. Analysis of kinetic parameters revealed that dextranase performs optimum cleavage of low molecular weight dextran (5000 Da, 0.5%) at 35ºC in 15 min at pH 4.5 with a Km and Vmax of 0.3738 mg/ml and 182.0 µmol/min, respectively. Thermal stability profiling of dextranase showed that it retained 80% activity up to 6 hours at 30-35ºC and remains 90% active at pH 4.5. In short, the dextranase reported here performs rapid cleavage of substrate at mild operational conditions which makes it an ideal candidate for dextran removal in sugar processing industry and for commercial production of low molecular weight oligosaccharides.

Keywords: Bacillus licheniformis, dextranase, gel permeation chromatograpy, enzyme purification, enzyme kinetics

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248 Designing Function Knitted and Woven Upholstery Textile With SCOPY Film

Authors: Manar Y. Abd El-Aziz, Alyaa E. Morgham, Amira A. El-Fallal, Heba Tolla E. Abo El Naga

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Different textile materials are usually used in upholstery. However, upholstery parts may become unhealthy when dust accrues and bacteria raise on the surface, which negatively affects the user's health. Also, leather and artificial leather were used in upholstery but, leather has a high cost and artificial leather has a potential chemical risk for users. Researchers have advanced vegie leather made from bacterial cellulose a symbiotic culture of bacteria and yeast (SCOBY). SCOBY remains a gelatinous, cellulose biofilm discovered floating at the air-liquid interface of the container. But this leather still needs some enhancement for its mechanical properties. This study aimed to prepare SCOBY, produce bamboo rib knitted fabrics with two different stitch densities, and cotton woven fabric then laminate these fabrics with the prepared SCOBY film to enhance the mechanical properties of the SCOBY leather at the same time; add anti-microbial function to the prepared fabrics. Laboratory tests were conducted on the produced samples, including tests for function properties; anti-microbial, thermal conductivity and light transparency. Physical properties; thickness and mass per unit. Mechanical properties; elongation, tensile strength, young modulus, and peel force. The results showed that the type of the fabric affected significantly SCOBY properties. According to the test results, the bamboo knitted fabric with higher stitch density laminated with SCOBY was chosen for its tensile strength and elongation as the upholstery of a bed model with antimicrobial properties and comfortability in the headrest design. Also, the single layer of SCOBY was chosen regarding light transparency and lower thermal conductivity for the creation of a lighting unit built into the bed headboard.

Keywords: anti-microbial, bamboo, rib, SCOPY, upholstery

Procedia PDF Downloads 58
247 Fructose-Aided Cross-Linked Enzyme Aggregates of Laccase: An Insight on Its Chemical and Physical Properties

Authors: Bipasa Dey, Varsha Panwar, Tanmay Dutta

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Laccase, a multicopper oxidase (EC 1.10.3.2) have been at the forefront as a superior industrial biocatalyst. They are versatile in terms of bestowing sustainable and ecological catalytic reactions such as polymerisation, xenobiotic degradation and bioremediation of phenolic and non-phenolic compounds. Regardless of the wide biotechnological applications, the critical limiting factors viz. reusability, retrieval, and storage stability still prevail. This can cause an impediment in their applicability. Crosslinked enzyme aggregates (CLEAs) have emerged as a promising technique that rehabilitates these essential facets, albeit at the expense of their enzymatic activity. The carrier free crosslinking method prevails over the carrier-bound immobilisation in conferring high productivity, low production cost owing to the absence of additional carrier and circumvent any non-catalytic ballast which could dilute the volumetric activity. To the best of our knowledge, the ε-amino group of lysyl residue is speculated as the best choice for forming Schiff’s base with glutaraldehyde. Despite being most preferrable, excess glutaraldehyde can bring about disproportionate and undesirable crosslinking within the catalytic site and hence could deliver undesirable catalytic losses. Moreover, the surface distribution of lysine residues in Trametes versicolor laccase is significantly less. Thus, to mitigate the adverse effect of glutaraldehyde in conjunction with scaling down the degradation or catalytic loss of the enzyme, crosslinking with inert substances like gelatine, collagen, Bovine serum albumin (BSA) or excess lysine is practiced. Analogous to these molecules, sugars have been well known as a protein stabiliser. It helps to retain the structural integrity, specifically secondary structure of the protein during aggregation by changing the solvent properties. They are comprehended to avert protein denaturation or enzyme deactivation during precipitation. We prepared crosslinked enzyme aggregates (CLEAs) of laccase from T. versicolor with the aid of sugars. The sugar CLEAs were compared with the classic BSA and glutaraldehyde laccase CLEAs concerning physico-chemical properties. The activity recovery for the fructose CLEAs were found to be ~20% higher than the non-sugar CLEA. Moreover, the 𝐾𝑐𝑎𝑡𝐾𝑚⁄ values of the CLEAs were two and three-fold higher than BSA-CLEA and GACLEA, respectively. The half-life (t1/2) deciphered by sugar-CLEA was higher than the t1/2 of GA-CLEAs and free enzyme, portraying more thermal stability. Besides, it demonstrated extraordinarily high pH stability, which was analogous to BSA-CLEA. The promising attributes of increased storage stability and recyclability (>80%) gives more edge to the sugar-CLEAs over conventional CLEAs of their corresponding free enzyme. Thus, sugar-CLEA prevails in furnishing the rudimentary properties required for a biocatalyst and holds many prospects.

Keywords: cross-linked enzyme aggregates, laccase immobilization, enzyme reusability, enzyme stability

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246 Separating Landform from Noise in High-Resolution Digital Elevation Models through Scale-Adaptive Window-Based Regression

Authors: Anne M. Denton, Rahul Gomes, David W. Franzen

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High-resolution elevation data are becoming increasingly available, but typical approaches for computing topographic features, like slope and curvature, still assume small sliding windows, for example, of size 3x3. That means that the digital elevation model (DEM) has to be resampled to the scale of the landform features that are of interest. Any higher resolution is lost in this resampling. When the topographic features are computed through regression that is performed at the resolution of the original data, the accuracy can be much higher, and the reported result can be adjusted to the length scale that is relevant locally. Slope and variance are calculated for overlapping windows, meaning that one regression result is computed per raster point. The number of window centers per area is the same for the output as for the original DEM. Slope and variance are computed by performing regression on the points in the surrounding window. Such an approach is computationally feasible because of the additive nature of regression parameters and variance. Any doubling of window size in each direction only takes a single pass over the data, corresponding to a logarithmic scaling of the resulting algorithm as a function of the window size. Slope and variance are stored for each aggregation step, allowing the reported slope to be selected to minimize variance. The approach thereby adjusts the effective window size to the landform features that are characteristic to the area within the DEM. Starting with a window size of 2x2, each iteration aggregates 2x2 non-overlapping windows from the previous iteration. Regression results are stored for each iteration, and the slope at minimal variance is reported in the final result. As such, the reported slope is adjusted to the length scale that is characteristic of the landform locally. The length scale itself and the variance at that length scale are also visualized to aid in interpreting the results for slope. The relevant length scale is taken to be half of the window size of the window over which the minimum variance was achieved. The resulting process was evaluated for 1-meter DEM data and for artificial data that was constructed to have defined length scales and added noise. A comparison with ESRI ArcMap was performed and showed the potential of the proposed algorithm. The resolution of the resulting output is much higher and the slope and aspect much less affected by noise. Additionally, the algorithm adjusts to the scale of interest within the region of the image. These benefits are gained without additional computational cost in comparison with resampling the DEM and computing the slope over 3x3 images in ESRI ArcMap for each resolution. In summary, the proposed approach extracts slope and aspect of DEMs at the lengths scales that are characteristic locally. The result is of higher resolution and less affected by noise than existing techniques.

Keywords: high resolution digital elevation models, multi-scale analysis, slope calculation, window-based regression

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245 Antimicrobial, Antioxidant and Cytotoxic Activities of Cleoma viscosa Linn. Crude Extracts

Authors: Suttijit Sriwatcharakul

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

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

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244 On Stochastic Models for Fine-Scale Rainfall Based on Doubly Stochastic Poisson Processes

Authors: Nadarajah I. Ramesh

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Much of the research on stochastic point process models for rainfall has focused on Poisson cluster models constructed from either the Neyman-Scott or Bartlett-Lewis processes. The doubly stochastic Poisson process provides a rich class of point process models, especially for fine-scale rainfall modelling. This paper provides an account of recent development on this topic and presents the results based on some of the fine-scale rainfall models constructed from this class of stochastic point processes. Amongst the literature on stochastic models for rainfall, greater emphasis has been placed on modelling rainfall data recorded at hourly or daily aggregation levels. Stochastic models for sub-hourly rainfall are equally important, as there is a need to reproduce rainfall time series at fine temporal resolutions in some hydrological applications. For example, the study of climate change impacts on hydrology and water management initiatives requires the availability of data at fine temporal resolutions. One approach to generating such rainfall data relies on the combination of an hourly stochastic rainfall simulator, together with a disaggregator making use of downscaling techniques. Recent work on this topic adopted a different approach by developing specialist stochastic point process models for fine-scale rainfall aimed at generating synthetic precipitation time series directly from the proposed stochastic model. One strand of this approach focused on developing a class of doubly stochastic Poisson process (DSPP) models for fine-scale rainfall to analyse data collected in the form of rainfall bucket tip time series. In this context, the arrival pattern of rain gauge bucket tip times N(t) is viewed as a DSPP whose rate of occurrence varies according to an unobserved finite state irreducible Markov process X(t). Since the likelihood function of this process can be obtained, by conditioning on the underlying Markov process X(t), the models were fitted with maximum likelihood methods. The proposed models were applied directly to the raw data collected by tipping-bucket rain gauges, thus avoiding the need to convert tip-times to rainfall depths prior to fitting the models. One advantage of this approach was that the use of maximum likelihood methods enables a more straightforward estimation of parameter uncertainty and comparison of sub-models of interest. Another strand of this approach employed the DSPP model for the arrivals of rain cells and attached a pulse or a cluster of pulses to each rain cell. Different mechanisms for the pattern of the pulse process were used to construct variants of this model. We present the results of these models when they were fitted to hourly and sub-hourly rainfall data. The results of our analysis suggest that the proposed class of stochastic models is capable of reproducing the fine-scale structure of the rainfall process, and hence provides a useful tool in hydrological modelling.

Keywords: fine-scale rainfall, maximum likelihood, point process, stochastic model

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243 Functional Significance of Qatari Camels Milk: Antioxidant Content and Antimicrobial Activity of Protein Fractions

Authors: Tahra ElObeid, Omnya Ahmed, Reem Al-Sharshani, Doaa Dalloul, Jannat Alnattei

Abstract:

Background: Camelus dormedarius camels are also called ‘the Arabian camels’ and are present in the desert area of North Africa and the Middle East. Recently, camel’s milk has a great attention globally because of their proteins and peptides that have been reported to be beneficial for the health and in the management of many diseases. Objectives: This study was designed to investigate the antioxidant, antimicrobial activity and to evaluate the total phenolic content of camel’s milk proteins in Qatar. Methods: Fresh two camel’s milk samples from Omani breed and called Muhajer (camel’s milk A and B) were collected on the 1st of the December. Both samples were from the same location Al- Shahaniyah, Doha, Qatar, but from different local private farms and feeding system. Camel’s milk A and B were defatted by centrifugation and their proteins were extracted by acid and thermal precipitation. The antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Total phenolic compound (TPC) was evaluated by Folin-Ciocalteu reagent (FCR). On the other hand, the antimicrobial activity against eight different type of pathogenic bacteria was evaluated by disc diffusion method and the zone of inhibition was measured. Results: The of the total phenolic content of whole milk in both camel’s milk A and B were significantly the highest among the protein extracts. The % of the DPPH radical inhibition of casein protein in both camel’s milk A and B were significantly the highest among the protein extracts. In this study, there were marked changes in the antibacterial activity in the different camel milk protein extracts. All extracts showed bacterial overgrowth. Conclusion: The antioxidant activity of the camel milk protein extracts correlated to their unique phenolic compounds and bioactive protein peptides. The antimicrobial activity was not detected perhaps due to the technique, the quality, or the extraction method. Overall, camel's milk exhibits a high antioxidant activity, which is responsible for many health benefits besides the nutritional values.

Keywords: camels milk, antioxidant content, antimicrobial activity, proteins, Qatar

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242 Non-Candida Albicans Candida: Virulence Factors and Species Identification in India

Authors: Satender Saraswat, Dharmendra Prasad Singh, Rajesh Kumar Verma, Swati Sarswat

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Background and Purpose: The predominant cause of candidiasis was Candida albicans which has shifted towards non-Candida albicans Candida (NCAC) (Candida species other than the C. albicans). NCAC, earlier considered non-pathogenic or minimally virulent, are now considered a primary cause of morbidity and mortality in immunocompromised. With the NCAC spp. gaining weightage in the clinical cases, this study was conducted to determine the prevalence of NCAC spp. in different clinical specimens and to assess a few of their virulence factors. Material and Methods: Routine samples for bacterial culture and sensitivity, showing colony characteristics like Candida on Blood Agar and microscopic features resembling Candida spp. were processed further. Candida isolates were tested for chlamydospore formation, biochemical tests including sugar fermentation and sugar assimilation tests, and growth at 42oC, colony colour on HiCrome™ Candida Differential Agar, HiCandida Identification Kit and VITEK-2 Compact. Virulence factors like adherence to buccal epithelial cells (ABEC), biofilm formation, hemolytic activity, and production of coagulase enzyme were also tested. Results: Mean age of the patients was 38.46 with a male-female ratio of 1.36:1. 137 Candida isolates were recovered. 45.3% isolates were isolated from urine, 19.7% from vaginal swabs and 13.9% from oropharyngeal swabs. 55 (40.1%) isolates of C. albicans and 82 (59.9%) of NCAC spp. were identified, with C. tropicalis (23.4%) in NCAC. C. albicans (3; 50%) was the commonest species in cases of candidemia. Haemolysin production (85.5%) and ABEC (78.2%) were the major virulence factors in C. albicans. C. tropicalis (59.4%) and C. dubliniensis (50%) showed maximum ABEC. Biofilm forming capacity was higher in C. tropicalis (78.1%) than C. albicans (67%). Conclusion: This study suggests varied prevalence and virulence based on geographical locations, even within a subcontinent. It clearly demarcates the emergence of NCAC and their predominance in different body fluids. Identification of Candida to species level should become a routine in all the laboratories.

Keywords: ABEC, NCAC, non-Candida albicans Candida, Vitek-2TM compact

Procedia PDF Downloads 126