Search results for: bacterial abundance

Authors: R. Sanchez-Salcedo, N. H. Voelcker

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Currently, low breast milk production in women is one of the leading health complications in infants. Recently, It has been demonstrated that exclusive breastfeeding, especially up to a minimum of 6 months, significantly reduces respiratory and gastrointestinal infections, which are the main causes of death in infants. However, the current data shows that a high percentage of women stop breastfeeding their children because they perceive an inadequate supply of milk, and only 45% of children are breastfeeding under 6 months. It is, therefore, clear the necessity to design and develop a biosensor that is sensitive and selective enough to identify and validate a panel of milk biomarkers that allow the early diagnosis of this condition. In this context, electrochemical biosensors could be a powerful tool for assessing all the requirements in terms of reliability, selectivity, sensitivity, cost efficiency and potential for multiplex detection. Moreover, they are suitable for the development of POC devices and wearable sensors. In this work, we report the development of two types of sensing platforms towards several biomarkers, including miRNAs and hormones present in breast milk and dysregulated in this pathological condition. The first type of sensing platform consists of an enzymatic sensor for the detection of lactose, one of the main components in milk. In this design, we used gold surface as an electrochemical transducer due to the several advantages, such as the variety of strategies available for its rapid and efficient functionalization with bioreceptors or capture molecules. For the second type of sensing platform, nanoporous silicon film (pSi) was chosen as the electrode material for the design of DNA sensors and aptasensors targeting miRNAs and hormones, respectively. pSi matrix offers a large superficial area with an abundance of active sites for the immobilization of bioreceptors and tunable characteristics, which increase the selectivity and specificity, making it an ideal alternative material. The analytical performance of the designed biosensors was not only characterized in buffer but also validated in minimally treated breastmilk samples. We have demonstrated the potential of an electrochemical transducer on pSi and gold surface for monitoring clinically relevant biomarkers associated with the heightened risk of low milk production in women. This approach, in which the nanofabrication techniques and the functionalization methods were optimized to increase the efficacy of the biosensor highly provided a foundation for further research and development of targeted diagnosis strategies.

Keywords: biosensors, electrochemistry, early diagnosis, clinical markers, miRNAs

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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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Authors: Mariam Dianat Sabet Gilani, Lars M. Blank, Birgitta E. Ebert

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Plant-derived lupane-type, pentacyclic triterpenoids are biologically active compounds that are highly interesting for applications in medical, pharmaceutical, and cosmetic industries. Due to the low abundance of these valuable compounds in their natural sources, and the environmentally harmful downstream process, alternative production methods, such as microbial cell factories, are investigated. Engineered Saccharomyces cerevisiae strains, harboring the heterologous genes for betulinic acid synthesis, can produce up to 2 g L-1 triterpenoids, showing high potential for large-scale production of triterpenoids. One limitation of the microbial synthesis is the intracellular product accumulation. It not only makes cell disruption a necessary step in the downstream processing but also limits productivity and product yield per cell. To overcome these restrictions, the aim of this study is to develop an in-situ extraction method, which extracts triterpenoids into a second organic phase. Such a continuous or sequential product removal from the biomass keeps the cells in an active state and enables extended production time or biomass recycling. After screening of twelve different solvents, selected based on product solubility, biocompatibility, as well as environmental and health impact, isopropyl myristate (IPM) was chosen as a suitable solvent for in-situ product removal from S. cerevisiae. Impedance-based single-cell analysis and off-gas measurement of carbon dioxide emission showed that cell viability and physiology were not affected by the presence of IPM. Initial experiments demonstrated that after the addition of 20 vol % IPM to cultures in the stationary phase, 40 % of the total produced triterpenoids were extracted from the cells into the organic phase. In future experiments, the application of IPM in a repeated batch process will be tested, where IPM is added at the end of each batch run to remove triterpenoids from the cells, allowing the same biocatalysts to be used in several sequential batch steps. Due to its high biocompatibility, the amount of IPM added to the culture can also be increased to more than 20 vol % to extract more than 40 % triterpenoids in the organic phase, allowing the cells to produce more triterpenoids. This highlights the potential for the development of a continuous large-scale process, which allows biocatalysts to produce intracellular products continuously without the necessity of cell disruption and without limitation of the cell capacity.

Keywords: betulinic acid, biocompatible solvent, in-situ extraction, isopropyl myristate, process development, secondary metabolites, triterpenoids, yeast

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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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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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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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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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Authors: Susan Mwai, Mary Muchane, Peter Wachira, Sheila Okoth, Muchai Muchane, Halima Saado

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Tropical forests harbor a wide range of biodiversity and rich macro-fungi diversity compared to the temperate regions in the World. However, biodiversity is facing the threat of extinction following the rate of forest loss taking place before proper study and documentation of macrofungi is achieved. The present study was undertaken to determine the effect of converting indigenous habitat to plantation forest on macrofungi diversity. To achieve the objective of this study, an inventory focusing on macro-fungi diversity was conducted within Kereita block in Kikuyu Escarpment forest which is on the southern side of Aberdare mountain range. The macrofungi diversity was conducted in the indigenous forest and in more than 15 year old Patula plantation forest , during the wet (long rain season, December 2014) and dry (Short rain season, May, 2015). In each forest type, 15 permanent (20m x 20m) sampling plots distributed across three (3) forest blocks were used. Both field and laboratory methods involved recording abundance of fruiting bodies, taxonomic identity of species and analysis of diversity indices and measures in terms of species richness, density and diversity. R statistical program was used to analyze for species diversity and Canoco 4.5 software for species composition. A total number of 76 genera in 28 families and 224 species were encountered in both forest types. The most represented taxa belonged to the Agaricaceae (16%), Polyporaceae (12%), Marasmiaceae, Mycenaceae (7%) families respectively. Most of the recorded macro-fungi were saprophytic, mostly colonizing the litter 38% and wood 34% based substrates, which was followed by soil organic dwelling species (17%). Ecto-mycorrhiza fungi (5%) and parasitic fungi (2%) were the least encountered. The data established that indigenous forests (native ecosystems) hosts a wide range of macrofungi assemblage in terms of density (2.6 individual fruit bodies / m2), species richness (8.3 species / plot) and species diversity (1.49/ plot level) compared to the plantation forest. The Conversion of native forest to plantation forest also interfered with species composition though did not alter species diversity. Seasonality was also shown to significantly affect the diversity of macro-fungi and 61% of the total species being present during the wet season. Based on the present findings, forested ecosystems in Kenya hold diverse macro-fungi community which warrants conservation measures.

Keywords: diversity, Indigenous forest, macro-fungi, plantation forest, season

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Authors: Esmaeil Kouhgardi, Elaheh Shakerdargah

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The mangrove ecosystem is a complex of various inter-related elements in the land-sea interface zone which is linked with other natural systems of the coastal region such as corals, sea-grass, coastal fisheries and beach vegetation. The mangrove ecosystem consists of water, muddy soil, trees, shrubs, and their associated flora, fauna and microbes. It is a very productive ecosystem sustaining various forms of life. Its waters are nursery grounds for fish, crustacean, and mollusk and also provide habitat for a wide range of aquatic life, while the land supports a rich and diverse flora and fauna, but pollutions may affect these characteristics. Iran has the lowest share of Persian Gulf pollution among the eight littoral states; environmental experts are still deeply concerned about the serious consequences of the pollution in the oil-rich gulf. Prolongation of critical conditions in the Persian Gulf has endangered its aquatic ecosystem. Water purification equipment, refineries, wastewater emitted by onshore installations, especially petrochemical plans, urban sewage, population density and extensive oil operations of Arab states are factors contaminating the Persian Gulf waters. Population density has been the major cause of pollution and environmental degradation in the Persian Gulf. Persian Gulf is a closed marine environment which is connected to open waterways only from one way. It usually takes between three and four years for the gulf's water to be completely replaced. Therefore, any pollution entering the water will remain there for a relatively long time. Presently, the high temperature and excessive salt level in the water have exposed the marine creatures to extra threats, which mean they have to survive very tough conditions. The natural environment of the Persian Gulf is very rich with good fish grounds, extensive coral reefs and pearl oysters in abundance, but has become increasingly under pressure due to the heavy industrialization and in particular the repeated major oil spillages associated with the various recent wars fought in the region. Pollution may cause the mortality of mangrove forests by effect on root, leaf and soil of the area. Study was showed the high correlation between industrial pollution and mangrove forests health in south of Iran and increase of population, coupled with economic growth, inevitably caused the use of mangrove lands for various purposes such as construction of roads, ports and harbors, industries and urbanization.

Keywords: Mangrove forest, pollution, Persian Gulf, population, environment

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Authors: Jeremy Jenrette, Z. Y. C. Liu, Pranav Chimote, Edward Fox, Trevor Hastie, Francesco Ferretti

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Suitable shark conservation depends on well-informed population assessments. Direct methods such as scientific surveys and fisheries monitoring are adequate for defining population statuses, but species-specific indices of abundance and distribution coming from these sources are rare for most shark species. We can rapidly fill these information gaps by boosting media-based remote monitoring efforts with machine learning and automation. We created a database of shark images by sourcing 24,546 images covering 219 species of sharks from the web application spark pulse and the social network Instagram. We used object detection to extract shark features and inflate this database to 53,345 images. We packaged object-detection and image classification models into a Shark Detector bundle. We developed the Shark Detector to recognize and classify sharks from videos and images using transfer learning and convolutional neural networks (CNNs). We applied these models to common data-generation approaches of sharks: boosting training datasets, processing baited remote camera footage and online videos, and data-mining Instagram. We examined the accuracy of each model and tested genus and species prediction correctness as a result of training data quantity. The Shark Detector located sharks in baited remote footage and YouTube videos with an average accuracy of 89\%, and classified located subjects to the species level with 69\% accuracy (n =\ eight species). The Shark Detector sorted heterogeneous datasets of images sourced from Instagram with 91\% accuracy and classified species with 70\% accuracy (n =\ 17 species). Data-mining Instagram can inflate training datasets and increase the Shark Detector’s accuracy as well as facilitate archiving of historical and novel shark observations. Base accuracy of genus prediction was 68\% across 25 genera. The average base accuracy of species prediction within each genus class was 85\%. The Shark Detector can classify 45 species. All data-generation methods were processed without manual interaction. As media-based remote monitoring strives to dominate methods for observing sharks in nature, we developed an open-source Shark Detector to facilitate common identification applications. Prediction accuracy of the software pipeline increases as more images are added to the training dataset. We provide public access to the software on our GitHub page.

Keywords: classification, data mining, Instagram, remote monitoring, sharks

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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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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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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

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

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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

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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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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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Authors: Cliff Shunsheng Han

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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

Procedia PDF Downloads 131

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

Procedia PDF Downloads 440

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

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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

Procedia PDF Downloads 272

Authors: Zafar Iqbal Bhatti

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The present research aims to examine and evaluate the frequencies and practices of verb complementation patterns in English newspaper social columns published in Pakistan and Britain. The research will demonstrate that Pakistani English is a non-native variety of English having its own unique usual and logical characteristics, affected by way of the native languages and the culture, upon syntactic levels, making the variety users aware that any differences from British or American English that are systematic and regular, or another English language, are not even if they are unique, erroneous forms and typical characteristics of several kinds. The objectives are to examine the verb complementation patterns that British and Pakistani social columnists use in relation to their syntactic categories. Secondly, to compare the verb complementation patterns used in Pakistani and British English newspapers social columns. This study will figure out various verb complementation patterns in Pakistani and British English newspaper social columns and their occurrence and distribution. The word classes express different functions of words, such as action, event, or state of being. This research aims to evaluate whether there are any appreciable differences in the verb complementation patterns used in Pakistani and British English newspaper social columns. The results will show the number of varieties of verb complementation patterns in selected English newspapers social columns. This study will fill the gap of previous studies conducted in this field as they only explore a little about the differences between Pakistani and British English newspapers. It will also figure out a variety of languages used in Pakistani and British English journals, as well as regional and cultural values and variations. The researcher will use AntConc software in this study to extract the data for analysis. The researcher will use a concordance tool to identify verb complementation patterns in selected data. Then the researcher will manually categorize them because the same type of adverb can sometimes be used for various purposes. From 1st June 2022 to 30th Sep. 2022, a four-month written corpus of the social columns of PE and BE newspapers will be collected and analyzed. For the analysis of the research questions, 50 social columns will be selected from Pakistani newspapers and 50 from British newspapers. The researcher will collect a representative sample of data from Pakistani and British English newspaper social columns. The researcher will manually analyze the complementation patterns of each verb in each sentence, and then the researcher will determine how frequently each pattern occurs. The researcher will use syntactic characteristics of the verb complementation elements according to the description by Downing and Locke (2006). The researcher will examine all of the verb complementation patterns in the data, and the frequency and distribution of each verb complementation pattern will be evaluated using the software. The researcher will explore every possible verb complementation pattern in Pakistani and British English before calculating the occurrence and abundance of each verb pattern. The researcher will explore every possible verb complementation pattern in Pakistani English before calculating the frequency and distribution of each pattern.

Keywords: verb complementation, syntactic categories, newspaper social columns, corpus

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

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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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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 133

Authors: Mehrnaz Mostafavi

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Recent advances in single-molecule protein identification (ID) and quantification techniques are poised to revolutionize proteomics, enabling researchers to delve into single-cell proteomics and identify low-abundance proteins crucial for biomedical and clinical research. This paper introduces a different approach to single-molecule protein ID and quantification using tri-color amino acid tags and a plasmonic nanopore device. A comprehensive simulator incorporating various physical phenomena was designed to predict and model the device's behavior under diverse experimental conditions, providing insights into its feasibility and limitations. The study employs a whole-proteome single-molecule identification algorithm based on convolutional neural networks, achieving high accuracies (>90%), particularly in challenging conditions (95–97%). To address potential challenges in clinical samples, where post-translational modifications affecting labeling efficiency, the paper evaluates protein identification accuracy under partial labeling conditions. Solid-state nanopores, capable of processing tens of individual proteins per second, are explored as a platform for this method. Unlike techniques relying solely on ion-current measurements, this approach enables parallel readout using high-density nanopore arrays and multi-pixel single-photon sensors. Convolutional neural networks contribute to the method's versatility and robustness, simplifying calibration procedures and potentially allowing protein ID based on partial reads. The study also discusses the efficacy of the approach in real experimental conditions, resolving functionally similar proteins. The theoretical analysis, protein labeler program, finite difference time domain calculation of plasmonic fields, and simulation of nanopore-based optical sensing are detailed in the methods section. The study anticipates further exploration of temporal distributions of protein translocation dwell-times and the impact on convolutional neural network identification accuracy. Overall, the research presents a promising avenue for advancing single-molecule protein identification and quantification with broad applications in proteomics research. The contributions made in methodology, accuracy, robustness, and technological exploration collectively position this work at the forefront of transformative developments in the field.

Keywords: nano proteomics, nanopore-based optical sensing, deep learning, artificial intelligence

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Authors: Lorenzo Petrucci

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This paper is intended to give insight into the potential use of small-scale off-grid trigeneration systems powered by biogas generated in a dairy farm. The off-grid plant object of analysis comprises a dual-fuel Genset as well as electrical and thermal storage equipment and an adsorption machine. The loads are the different apparatus used in the dairy farm, a household where the workers live and a small electric vehicle whose batteries can also be used as a power source in case of emergency. The insertion in the plant of an adsorption machine is mainly justified by the abundance of thermal energy and the simultaneous high cooling demand associated with the milk-chilling process. In the evaluated operational scenario, our research highlights the importance of prioritizing specific small loads which cannot sustain an interrupted supply of power over time. As a consequence, a photovoltaic and thermal panel is included in the plant and is tasked with providing energy independently of potentially disruptive events such as engine malfunctioning or scarce and unstable supplies of fuels. To efficiently manage the plant an energy dispatch strategy is created in order to control the flow of energy between the power sources and the thermal and electric storages. In this article we elaborate on models of the equipment and from these models, we extract parameters useful to build load-dependent profiles of the prime movers and storage efficiencies. We show that under reasonable assumptions the analysis provides a sensible estimate of the generated energy. The simulations indicate that a Diesel Generator sized to a value 25% higher than the total electrical peak demand operates 65% of the time below the minimum acceptable load threshold. To circumvent such a critical operating mode, dump loads are added through the activation and deactivation of small resistors. In this way, the excess of electric energy generated can be transformed into useful heat. The combination of PVT and electrical storage to support the prioritized load in an emergency scenario is evaluated in two different days of the year having the lowest and highest irradiation values, respectively. The results show that the renewable energy component of the plant can successfully sustain the prioritized loads and only during a day with very low irradiation levels it also needs the support of the EVs’ battery. Finally, we show that the adsorption machine can reduce the ice builder and the air conditioning energy consumption by 40%.

Keywords: hybrid power plants, mathematical modeling, off-grid plants, renewable energy, trigeneration

Procedia PDF Downloads 174

Authors: Luz del Carmen Rubí Félix Peña, Jose Adan Felix-Ortiz, Ely Sara Lopez-Alvarez, Wenceslao Valenzuela-Quiñonez

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On a global scale, Mexico is the sixth largest producer of farmed white shrimp (Penaeus vannamei). The activity suffered significant economic losses due to acute hepatopancreatic necrosis (AHPND) caused by a strain of Vibrio parahaemolyticus. For control, the first option is the application of antibiotics in food, causing changes in the environment and bacterial communities, which has produced greater virulence and resistance of pathogenic bacteria. An alternative treatment is silver nanoparticles (AgNPs) generated by green synthesis, which have shown an antibacterial capacity by destroying the cell membrane or denaturing the cell. However, the doses at which these are effective are still unknown. The aim is to calculate the minimum inhibitory concentration (MIC) using the Gompertz, Richard, and Logistic model of biosynthesized AgNPs against a strain of V. parahaemolyticus. Through the testing of different formulations of AgNPs synthesized from Euphorbia prostrate (Ep) extracts against V. parahaemolyticus causing AHPND in white shrimp. Aqueous and ethanol extracts were obtained, and the concentration of phenols and flavonoids was quantified. In the antibiograms, AgNPs were formulated in ethanol extracts of Ep (20 and 30%). The inhibition halo at well dilution test were 18±1.7 and 17.67±2.1 mm against V. parahaemolyticus. A broth microdilution was performed with the inhibitory agents (aqueous and ethanolic extracts and AgNPs) and 20 μL of the inoculum of V. parahaemolyticus. The MIC for AgNPs was 6.2-9.3 μg/mL and for ethanol extract of 49-73 mg/mL. The Akaike index (AIC) was used to choose the Gompertz model for ethanol extracts of Ep as the best data descriptor (AIC=204.8, 10%; 45.5, 20%, and 204.8, 30%). The Richards model was at AgNPs ethanol extract with AIC=-9.3 (10%), -17.5 (20 and 30%). The MIC calculated for EP extracts with the modified Gompertz model were 20 mg/mL (10% and 20% extract) and 40 mg/mL at 30%, while Richard was winner for AgNPs-synthesized it was 5 μg/mL (10% and 20%) and 8 μg/mL (30%). The solver tool Excel was used for the calculations of the models and inhibition curves against V.parahaemolyticus.

Keywords: green synthesis, euphorbia prostata, phenols, flavonoids, bactericide

Procedia PDF Downloads 106

Authors: Marta Skwarecka, Patrycja Bloch, Rafal Walkusz, Oliwia Urbanowicz, Grzegorz Zielinski, Sabina Zoledowska, Dawid Nidzworski

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Upper respiratory tract infections are one of the most common reasons for visiting a general doctor. Streptococci are the most common bacterial etiological factors in these infections. There are many different types of Streptococci and infections vary in severity from mild throat infections to pneumonia. For example, S. pyogenes mainly contributes to acute pharyngitis, palatine tonsils and scarlet fever, whereas S. Streptococcus pneumoniae is responsible for several invasive diseases like sepsis, meningitis or pneumonia with high mortality and dangerous complications. There are only a few diagnostic tests designed for detection Streptococci from the infected throat of patients. However, they are mostly based on lateral flow techniques, and they are not used as a standard due to their low sensitivity. The diagnostic standard is to culture patients throat swab on semi selective media in order to multiply pure etiological agent of infection and subsequently to perform antibiogram, which takes several days from the patients visit in the clinic. Therefore, the aim of our studies is to develop and implement to the market a Point of Care device for the rapid identification of Streptococcus pyogenes and Streptococcus pneumoniae with simultaneous identification of antibiotic resistance genes. In the course of our research, we successfully selected genes for to-species identification of Streptococci and genes encoding antibiotic resistance proteins. We have developed a reaction to amplify these genes, which allows detecting the presence of S. pyogenes or S. pneumoniae followed by testing their resistance to erythromycin, chloramphenicol and tetracycline. What is more, the detection of β-lactamase-encoding genes that could protect Streptococci against antibiotics from the ampicillin group, which are widely used in the treatment of this type of infection is also developed. The test is carried out directly from the patients' swab, and the results are available after 20 to 30 minutes after sample subjection, which could be performed during the medical visit.

Keywords: antibiotic resistance, Streptococci, respiratory infections, diagnostic test

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Authors: Uchenna Egwu, Paul Jonathan Sallis

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Perennial ryegrass is an abundant renewable lignocellulosic biofuel feedstock for biomethane production through anaerobic digestion (AD). In this study, six anaerobic continuously stirred tank reactors (CSTRs) were set up in three pairs. Each pair of the CSTRs was then used to study the effects of operating temperatures – psychrophilic, mesophilic, and thermophilic, and optimum pH on the specific methane production (SMP) of the ryegrass during AD under discontinuous daily feeding conditions. The reactors were fed at an organic loading rate (OLR) ranging from 1-1.5 kgVS.L⁻¹d⁻¹ and hydraulic residence time, HRT=20 days for 140 days. The pH of the digesters was maintained at the range of 6.8-7.2 using 1 M NH₄HCO₃ solution, but this was replaced with biomass ash-extracts from day 105-140. The results obtained showed that the mean SMP of ryegrass measured between HRT 3 and 4 were 318.4, 425.4 and 335 N L CH₄ kg⁻¹VS.d⁻¹ for the psychrophilic (25 ± 2°C), mesophilic (40 ± 1°C) and thermophilic (60 ± 1°C) temperatures respectively. It was also observed that the buffering ability of the reactors increased with operating temperature, probably due to an increase in the solubility of ammonium bicarbonate (NH₄HCO₃) with temperature. The reactors also achieved a mean VS destruction of 61.9, 68.5 and 63.5%, respectively, which signifies that the mesophilic reactors achieved the highest specific methane production (SMP), while the psychrophilic reactors achieved the lowest. None of the reactors attained steady-state condition due to the discontinuous daily feeding times, and therefore, such feeding practice may not be the most effective for maximum biogas production over long periods of time. The addition of NH₄HCO₃ as supplement provided a good buffering condition in these AD digesters, but the digesters failed in the long run due to inhibition from the accumulation of free ammonia, which later led to decrease in pH, acidification, and souring of the digesters. However, the addition of biomass ash extracts was shown to potentially revive failed AD reactors by providing an adequate buffering and essential trace nutrient supplements necessary for optimal bacterial growth.

Keywords: anaerobic digestion, discontinuous feeding, perennial ryegrass, specific methane production, supplements, temperature

Procedia PDF Downloads 127

Authors: Christopher Baiyegunhi, Kuiwu Liu, Oswald Gwavava

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Grain size analysis is a vital sedimentological tool used to unravel the hydrodynamic conditions, mode of transportation and deposition of detrital sediments. In this study, detailed grain-size analysis was carried out on thirty-five sandstone samples from the Ecca Group in the Eastern Cape Province of South Africa. Grain-size statistical parameters, bivariate analysis, linear discriminate functions, Passega diagrams and log-probability curves were used to reveal the depositional processes, sedimentation mechanisms, hydrodynamic energy conditions and to discriminate different depositional environments. The grain-size parameters show that most of the sandstones are very fine to fine grained, moderately well sorted, mostly near-symmetrical and mesokurtic in nature. The abundance of very fine to fine grained sandstones indicates the dominance of low energy environment. The bivariate plots that the samples are mostly grouped, except for the Prince Albert samples that show scattered trend, which is due to the either mixture of two modes in equal proportion in bimodal sediments or good sorting in unimodal sediments. The linear discriminant function (LDF) analysis is dominantly indicative of turbidity current deposits under shallow marine environments for samples from the Prince Albert, Collingham and Ripon Formations, while those samples from the Fort Brown Formation are fluvial (deltaic) deposits. The graphic mean value shows the dominance of fine sand-size particles, which point to relatively low energy conditions of deposition. In addition, the LDF results point to low energy conditions during the deposition of the Prince Albert, Collingham and part of the Ripon Formation (Pluto Vale and Wonderfontein Shale Members), whereas the Trumpeters Member of the Ripon Formation and the overlying Fort Brown Formation accumulated under high energy conditions. The CM pattern shows a clustered distribution of sediments in the PQ and QR segments, indicating that the sediments were deposited mostly by suspension and rolling/saltation, and graded suspension. Furthermore, the plots also show that the sediments are mainly deposited by turbidity currents. Visher diagrams show the variability of hydraulic depositional conditions for the Permian Ecca Group sandstones. Saltation is the major process of transportation, although suspension and traction also played some role during deposition of the sediments. The sediments were mainly in saltation and suspension before being deposited.

Keywords: grain size analysis, hydrodynamic condition, depositional environment, Ecca Group, South Africa

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Authors: Ferhat Mohamed Amine, Boukhatem Mohamed Nadjib, Chemat Farid

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The Lamiaceae family is widely spread in Algeria. Due to the application of Thymus species growing wild in Algeria as a culinary herb and in folk medicine, the purpose of the present work was to evaluate antimicrobial activities of their essential oils and relate them with their chemical composition, for further application in food and pharmaceutical industries as natural valuable products. The extraction of the Thymus vulgaris L. essential oil (TVEO) was obtained by steam distillation. Chemical composition of the TVEO was determined by Gas Chromatography. A total of thirteen compounds were identified. Carvacrol (83.8%) was the major component, followed by cymene (8.15%) and terpinene (4.96%). Antibacterial action of the TVEO against 23 clinically isolated bacterial strains was determined by using agar disc diffusion and vapour diffusion methods at different doses. By disc diffusion method, TVEO showed potent antimicrobial activity against gram-positive bacteria more than gram-negative strains and antibiotic discs. The Diameter of Inhibition Zone (DIZ) varied from 25 to 60 mm for S. aureus, B. subtilisand E. coli. However, the results obtained by both agar diffusion and vapour diffusion methods were different. Significantly higher antibacterial effect was observed in the vapour phase at lower doses. S. aureus and B. subtilis were the most susceptible strains to the oil vapour. Therefore, smaller doses of EO in the vapour phase can be inhibitory to pathogenic bacteria. There is growing evidence that TVEO in vapour phase are effective antiseptic systems and appears worthy to be considered for practical uses in the treatment of human infections oras air decontaminants in hospital. TVEO has considerable antibacterial activity deserving further investigation for clinical applications. Also whilst the mode of action remains mainly undetermined, this experimental approach will need to continue.

Keywords: antimicrobial drugs, carvacrol, disc diffusion, Thymus vulgaris, vapour diffusion

Procedia PDF Downloads 371