Search results for: lactic acid bacteria; multidrug-resistant pathogens

Authors: Inayat Ur Rahman Arshad

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Background: Stevia rebaudiana B. is a shrubby perennial herb of Asteraceae family that possesses the unique ability of accumulative non-caloric sweet steviol glycosides (SGs). Purpose: The purpose of the study is to optimize sugar concentration, pH level, and inoculum size for inducing the callus with optimum growth and efficient antibacterial potential. Method: Three different experiments were conducted in which Callus explant from three-months-old already established callus of Stevia reabudiana of four different sizes was inoculated on Murashige and Skoog (MS) basal medium supplemented with five different sucrose concentration and pH adjusted at four different levels. Results: Maximum callus induction 100, 87.5, and 85.33% resulted in the medium supplemented with 30 g/l sucrose, pH maintained at 5.5, and inoculated with 1.25g inoculum, respectively. Similarly, the highest fresh weights 65.00, 75.50, and 50.53 g/l were noted in a medium fortified with 40 g/l sucrose, inoculated 1.25g inoculum, and 6.0 pH level, respectively. However, the callus developed in a medium containing 50 g/l sucrose was found to be highly antibacterial potent with 27.3 and 26.5 mm inhibition zone against P. vulgaris and B. subtilis, respectively. Similarly, the callus grown on a medium inoculated with 1.00 g inoculum resulted in maximum antibacterial potential against S. aureus and P. vulgaris with 25 and 23.72 mm inhibition zone, respectively. However, in the case of pH levels, the medium maintained at 6.5 pH showed maximum antibacterial activity against P. vulgaris, B.subtilis, and E.coli with 27.9, 25, and 23.72 mm, respectively. The ethyl acetate extract of Stevia callus and leaves did not show antibacterial potential against Xanthomonas campestris and Clavebactor michiganensis. In the entire experiment, the standard antibacterial agent Streptomycin showed the highest inhibition zones among the rest of the callus extract; however, the pure dimethyl sulfoxide (DMSO) caused no inhibitory zone against any bacteria. Conclusion: From these findings, it is concluded that among various levels, sucrose @ 40 g L⁻¹, pH 6.0, and inoculums at 0.75 g were found best for most of the growth and quality attributes, including fresh weight, dry weight, and antibacterial activities and therefore can be recommended for callus proliferation and antibacterial potential of Stevia rebaudiana.

Keywords: Stevia rebaudiana, Steviol Glycosides, callus, Xanthomonas campestris

Procedia PDF Downloads 82

Authors: Pınar Karbuz, A. Seyhun Kıpcak, Mehmet B. Piskin, Emek Derun, Nurcan Tugrul

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Pectin is a complex colloidal polysaccharide, found on the cell walls of all young plants such as fruit and vegetables. It acts as a thickening, stabilizing and gelling agent in foods. Pectin was extracted from mandarin and lemon peels using ultrasound and microwave assisted extraction methods to compare with these two different sources and methods of pectin production. In this work, the effect of microwave power (360, 600 W) and irradiation time (1, 2, 3 min) on the yield of extracted pectin from mandarin and lemon peels for microwave assisted extraction (MAE) were investigated. For ultrasound assisted extraction (UAE), parameters were determined as temperature (60, 75 °C) and sonication time (15, 30, 45 min) and hydrochloric acid (HCl) was used as an extracting agent for both extraction methods. The highest yields of extracted pectin from lemon peels were found to be 8.16 % (w/w) for 75 °C, 45 min by UAE and 8.58 % (w/w) for 360 W, 1 min by MAE. Additionally, the highest yields of extracted pectin from mandarin peels were found to be 11.29 % (w/w) for 75 °C, 45 min by UAE and 16.44 % (w/w) for 600 W, 1 min by MAE. The results showed that the use of microwave assisted extraction promoted a better yield when compared to the two extraction methods. On the other hand, according to the results of experiments, mandarin peels contain more pectin than lemon peels when the compared to the pectin product values of two sources. Therefore, these results suggested that MAE could be used as an efficient and rapid method for extraction of pectin and mandarin peels should be preferred as sources of pectin production compared to lemon peels.

Keywords: mandarin peel, lemon peel, pectin, ultrasound, microwave, extraction

Procedia PDF Downloads 234

Authors: Siriniya Siribrahmanakul

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Introduction:SARsCOVID-19 is rapidly spreading, and some people may exhibit severe symptoms. Mortality rate of 2.0–3.0% with COVID-19 infection atworldwide. Human leukocyte antigen (HLA), located on chromosome 6, consist of HLA class I and class II. HLA are used by the immune system to attach self-antigens. Previous studies, HLA-DRB1*01:01,HLA-DRB1*12:01and HLA-DRB1*14:04 were, showed significant difference with severe COVID-19 in the Chinese population by p-value < 0.05. Objective: We investigated the prevalence of HLA-DRB1 alleles associated with severe COVID-19 in Thai population. Materials and Methods:200 DNA samples were isolated from EDTA blood using the MagNAprue Compact Nucleic Acid Isolation kits.HLA-DRB1alleles were genotyped using sequence-specific oligonucleotides (PCR-SSOs). Results:The frequency of HLA-DRB1 alleles in Thai population wereHLA-DRB1*12:02 (15.75%), HLA-DRB1*15:02 (14.50%), HLA-DRB1*09:01 (11.50%), HLA-DRB1*07:01 (9.50%), HLA-DRB1*03:01,HLA-DRB1*05:01 (5.75%), HLA-DRB1*14:01 (5.50%), HLA-DRB1*16:02 (4.50%), HLA-DRB1*04:05 (4.00%), HLA-DRB1*14:03 (3.25%), HLA-DRB1*10:01 (2.25%) and HLA-DRB1*13:02 (2.00%). Particularly, HLA-DRB1*12:02 allele was the highest allele frequency presented in the four regions groups of Thai population. Furthermore, the HLA-DRB1* alleles associated with severe COVID-19, which consists ofHLA-DRB1*14:04(2.00 %) and HLA-DRB1*12:01(0.50%) in Thai population, whereas HLA-DRB1*01:01 allele was not found in this population. HLA-DRB1*14:04 and HLA-DRB1*12:01alleles were similarly distributed in four regions populations in Thailand (p-value > 0.05). The alleles frequencies of HLA-DRB1*14:04 and HLA-DRB1*12:01, which associated with severe COVID-19, had no significant differences between Thai population, South China population, South Africa population, and South Koreapopulation (p-value > 0.05). Conclusions: Particularly, this study has focused on allele frequency of HLA-DRB1*14:04in a healthy Thai population to evaluating their impact on the severe COVID-19. Furthermore, our research needs to be done in larger numbers of Thai patients.

Keywords: HLA-DRB1, allele frequency, Thai population, COVID-19 marker

Procedia PDF Downloads 132

Authors: Cengiz Demir, Recep Keşli, Gülşah Aşık

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Objective: This study was carried out with the purpose of defining by using the E test method and determining the antibiotic resistance profiles of Gram-negative anaerobic bacilli isolated from various clinical specimens obtained from patients with suspected anaerobic infections and referred to Medical Microbiology Laboratory of Afyon Kocatepe University, ANS Application and Research Hospital. Methods: Two hundred and seventy eight clinical specimens were examined for isolation of the anaerobic bacteria in Medical Microbiology Laboratory between the 1st November 2014 and 30th October 2015. Specimens were cultivated by using Scheadler agar that 5% defibrinated sheep blood added, and Scheadler broth. The isolated anaerobic Gram-negative bacilli were identified conventional methods and Vitek 2 (ANC ID Card, bioMerieux, France) cards. Antibiotic resistance rates against to penicillin G, clindamycin, cefoxitin, metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem were determined with E-test method for each isolate. Results: Of the isolated twenty-eight anaerobic gram negative bacilli fourteen were identified as the B. fragilis group, 9 were Prevotella group, and 5 were Fusobacterium group. The highest resistance rate was found against penicillin (78.5%) and resistance rates against clindamycin and cefoxitin were found as 17.8% and 21.4%, respectively. Against to the; metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem, no resistance was found. Conclusion: Since high rate resistance has been detected against to penicillin in the study penicillin should not be preferred in empirical treatment. Cefoxitin can be preferred in empirical treatment; however, carrying out the antibiotic sensitivity testing will be more proper and beneficial. No resistance was observed against carbapenem group antibiotics and metronidazole; so that reason, these antibiotics should be reserved for treatment of infectious caused by resistant strains in the future.

Keywords: anaerobic gram-negative bacilli, anaerobe, antibiotics and resistance profiles, e-test method

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Authors: Saroj Poudel, Joshua Mancini, Douglas Pike, Jennifer Timm, Alexei Tyryshkin, Vikas Nanda, Paul Falkowski

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On the early Earth, protein-metal complexes likely harvested energy from a reduced environment. These complexes would have been precursors to the metabolic enzymes of ancient organisms. Hydrogenase is an essential enzyme in most anaerobic organisms for the reduction and oxidation of hydrogen in the environment and is likely one of the earliest evolved enzymes. To attempt to reinvent a precursor to modern hydrogenase, we computationally designed a short thirteen amino acid peptide that binds the often-required catalytic transition metal Nickel in hydrogenase. This simple complex can achieve hundreds of hydrogen evolution cycles using light energy in a broad range of temperature and pH. Biophysical and structural investigations strongly indicate the peptide forms a di-nickel active site analogous to Acetyl-CoA synthase, an ancient protein central to carbon reduction in the Wood-Ljungdahl pathway and capable of hydrogen evolution. This work demonstrates that prior to the complex evolution of multidomain enzymes, early peptide-metal complexes could have catalyzed energy transfer from the environment on the early Earth and enabled the evolution of modern metabolism

Keywords: hydrogenase, prebiotic enzyme, metalloenzyme, computational design

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Authors: Abdull Halim

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The Standard Penetration Test (SPT-N) is the most common in situ test for soil investigations. The Shearing Seismic Standard Penetration Test (SSPT-N), on the other hand, is a new method using shearing wave with propagation exponent equation between the shearing wave, Vs., and hardness, N values without any need for borehole data. Due to the fast and accurate results that can be obtained, the SSPT has found many applications such as in the field rectification buried pipe line, the acid tank settlement and foundation design analyses, and the quality control assessment. Many geotechnical regimes and properties have attempted to correlate both the SSPT and the SPT-N values. Various foundation design methods have been developed based on the outcomes of these tests. Hence, it is pertinent to correlate these tests so that either one of the test can be used in the absence of the other, especially for preliminary evaluation and design purposes. The primary purpose of this study was to investigate the relationship between the SSPT-N and SPT-N values for different types of cohesive soil in Peninsular Malaysia. Data were collected from four different sites, and the correlations were established between the hardness N values, principal stress-strain Mohr circle curve, cohesion, friction angle and vertical effective stress. A positive exponent relationship was found between the shearing wave, sVs., and the hardness N values of the soil. In general, the SSPT-N value was slightly lower than the SPT-N value due to the upper limit boundary of the soil layer.

Keywords: InsituSoil determination; shearing wave; hardness; correlation, SSPT-N, SPT-N

Procedia PDF Downloads 184

Authors: Li-Yu Lee, Yu-Shuan Chen, Jun Sheng Wang, I-Ming Chu

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Microneedle patch is a painless transdermal drug delivery method, It could solve some problems in traditional drug delivery such as digestive system causing drug metabolism and subcutaneous injection causing some side effects. Coating drug on or loading drug in microneedle can carry active ingredient through stratum corneum, also can control dose well when microneedle patch apply on localized topical area. We used hyaluronic acid to fabricate dissolvable microneedle patch and encapsulated minoxidil into microneedles. Minoxdil is a drug for exterior use that can be used to treat Androgenetic alopecia, but related commercial products have some shortcomings, for example, propylene glycol which is used to soften stratum corneum cause skin allergic reaction, comparing chemical promotion, microneedle patch provide physical way to make drugs through nature barrier of skin. In this research, we designed a two-step process to fabricate microneedle patch, that can effectively reduce drug waste, and gentle production process could maintain drug activity well. We also do in vitro test on cadaver to make sure patch has enough mechanical strength to penetrate stratum corneum. In the release test and animal test, we found microneedle patch has higher delivery efficiency than tradition way. In this study, we may determine that germinal MNs patch is a potential commodity.

Keywords: dissolving microneedles, androgenetic alopecia, minoxidil, transdermal drug delivery

Procedia PDF Downloads 279

Authors: Grace O. Babarinde, Adegoke O.Gabriel, Rahman Akinoso, Adekanye Bosede R.

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Health risks associated with the use of synthetic chemicals to control post-harvest losses in fruit calls for use of natural biodegradable compounds. The potential of Piper guineense as postharvest preservative for Roma tomato (Solanum lycopersicum L.) was investigated. Freshly harvested red tomato (200 g) was dipped into five concentrations (1, 2, 3, 4 and 5% w/v) of P. guineense aqueous extract, while untreated fruits served as control. The samples were stored under refrigeration and analysed at 5-day interval for physico-chemical properties. P. guineense essential oil (EO) was characterised using GC-MS and its tomato preservative potential was evaluated. Percentage weight loss (PWL) in extract-treated tomato ranged from 0.0-0.68% compared to control (0.3-19.97%) during storage. Values obtained for firmness ranged from 8.23-16.88 N and 8.4 N in extract-treated and control. pH reduced from 5.4 to 4.5 and 3.7 in extract-treated and untreated samples, respectively. Highest value of Total Soluble Solid (1.8 °Brix) and maximum retention of Ascorbic acid (13.0 mg/100 g) were observed in 4% P. guineense-treated samples. Predominant P. guineense EO components were zingiberene (9.9%), linalool (10.7%), β-caryophyllene (12.6%), 1, 5-Heptadiene, 6-methyl-2-(4-methyl-3-cyclohexene-l-yl) (16.4%) and β-sesquiphellandrene (23.7%). Tomatoes treated with EO had lower PWL (5.2%) and higher firmness (14.2 N) than controls (15.3% and 11.9 N) respectively. The result indicates that P. guineense can be incorporated in to post harvest technology of Roma tomato fruit.

Keywords: aqueous extract, essential oil, piper guineense, Roma tomato, storage condition

Procedia PDF Downloads 476

Authors: Lars A. L. Giske, Ola J. Mork, Emil Bjoerlykhaug

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The design of fish processing equipment greatly impacts how easy the cleaning process for the equipment is. This is a critical issue in fish processing, as cleaning of fish processing equipment is a task that is both costly and time consuming, in addition to being very important with regards to product quality. Even more, poorly cleaned equipment could in the worst case lead to contaminated product from which consumers could get ill. This paper will elucidate how equipment design changes could improve the work for the cleaners and saving money for the fish processing facilities by looking at a case for product design improvements. The design of fish processing equipment largely determines how easy it is to clean. “Design for cleaning” is the new hype in the industry and equipment where the ease of cleaning is prioritized gets a competitive advantage over equipment in which design for cleaning has not been prioritized. Design for cleaning is an important research area for equipment manufacturers. SeaSide AS is doing continuously improvements in the design of their products in order to gain a competitive advantage. The focus in this paper will be conveyors for internal logistic and a product called the “electro stunner” will be studied with regards to “Design for cleaning”. Often together with SeaSide’s customers, ideas for new products or product improvements are sketched out, 3D-modelled, discussed, revised, built and delivered. Feedback from the customers is taken into consideration, and the product design is revised once again. This loop was repeated multiple times, and led to new product designs. The new designs sometimes also cause the manufacturing processes to change (as in going from bolted to welded connections). Customers report back that the concrete changes applied to products by SeaSide has resulted in overall more easily cleaned equipment. These changes include, but are not limited to; welded connections (opposed to bolted connections), gaps between contact faces, opening up structures to allow cleaning “inside” equipment, and generally avoiding areas in which humidity and water may gather and build up. This is important, as there will always be bacteria in the water which will grow if the area never dries up. The work of creating more cleanable design is still ongoing, and will “never” be finished as new designs and new equipment will have their own challenges.

Keywords: cleaning, design, equipment, fish processing, innovation

Procedia PDF Downloads 237

Authors: Joao Almeida, Ana Azevedo, Myriam Kanoun-Boule, Maria Ines Santos, Antonio Tadeu

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The effects of the increasing expansion of cities and climate changes have encouraged European countries and regions to adopt nature-based solutions with ability to mitigate environmental issues and improve life in cities. Among these strategies, green roofs and urban gardens have been considered ingenious solutions, since they have the desirable potential to improve air quality, prevent floods, reduce the heat island effect and restore biodiversity in cities. However, an additional consumption of fresh water and mineral nutrients is necessary to sustain larger green urban areas. This communication discusses the main technical features of a new system to manage urine recovery at the building level and its application in green roofs. The depletion of critical nutrients like phosphorus constitutes an emergency. In turn, their elimination through urine is one of the principal causes for their loss. Thus, urine recovery in buildings may offer numerous advantages, constituting a valuable fertilizer abundantly available in cities and reducing the load on wastewater treatment plants. Although several urine-diverting toilets have been developed for this purpose and some experiments using urine directly in agriculture have already been carried out in Europe, several challenges have emerged with this practice concerning collection, sanitization, storage and application of urine in buildings. To our best knowledge, current buildings are not designed to receive these systems and integrated solutions with ability to self-manage the whole process of urine recovery, including separation, maturation and storage phases, are not known. Additionally, if from a hygiene point of view human urine may be considered a relatively safe fertilizer, the risk of disease transmission needs to be carefully analysed. A reduction in microorganisms can be achieved by storing the urine in closed tanks. However, several factors may affect this process, which may result in a higher survival rate for some pathogens. In this work, urine effluent was collected under real conditions, stored in closed containers and kept in climatic chambers under variable conditions simulating cold, temperate and tropical climates. These samples were subjected to a first physicochemical and microbiological control, which was repeated over time. The results obtained so far suggest that maturation conditions were reached for all the three temperatures and that a storage period of less than three months is required to achieve a strong depletion of microorganisms. The authors are grateful for the Project WashOne (POCI-01-0247-FEDER-017461) funded by the Operational Program for Competitiveness and Internationalization (POCI) of Portugal 2020, with the support of the European Regional Development Fund (FEDER).

Keywords: sustainable green roofs and urban gardens, urban nutrient cycle, urine-based fertilizers, urine recovery in buildings

Procedia PDF Downloads 166

Authors: H. Tayefih, F. farajzade ahari, F. Zarghami, V. Zeighamian, N. Zarghami, Y. Pilehvar-soltanahmadi

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Breast cancer is the most frequently occurring cancer among women throughout the world. Natural compounds such as curcumin hold promise to treat a variety of cancers including breast cancer. However, curcumin's therapeutic application is limited, due to its rapid degradation and poor aqueous solubility. On the other hand, previous studies have stated that drug delivery using nanoparticles might improve the therapeutic response to anticancer drugs. Poly (N-isopropylacrylamide-co-methacrylic acid) (PNIPAAm–MAA) is one of the hydrogel copolymers utilized in the drug delivery system for cancer therapy. The aim of this study was to examine the cytotoxic potential of curcumin encapsulated within the NIPAAm-MAA nanoparticle, on the MCF-7 breast cancer cell line. In this work, polymeric nanoparticles were synthesized through the free radical mechanism, and curcumin was encapsulated into NIPAAm-MAA nanoparticles. Then, the cytotoxic effect of curcumin-loaded NIPAAm-MAA on the MCF-7 breast cancer cell line was measured by MTT assays. The evaluation of the results showed that curcumin-loaded NIPAAm-MAA has more cytotoxic effect on the MCF-7 cell line and efficiently inhibited the growth of the breast cancer cell population, compared with free curcumin. In conclusion, this study indicates that curcumin-loaded NIPAAm-MAA suppresses the growth of the MCF-7 cell line. Overall, it is concluded that encapsulating curcumin into the NIPAAm-MAA copolymer could open up new avenues for breast cancer treatment.

Keywords: PNIPAAm-MAA, breast cancer, curcumin, drug delivery

Procedia PDF Downloads 374

Authors: Preet Singh, Kavitha Kongara, Dave Harding, Neil Ward, Paul Chambers

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The objective of this study was to compare the analgesic efficacy of opiorphin and its analogue with a mu-receptor agonist; morphine. Opiorphins (Gln-Arg-Phe-Ser-Arg) belong to the family of endogenous enkephalinase inhibitors, found in saliva of humans. They are inhibitors of two Zinc metal ectopeptidases (Neutral endopeptidase NEP, and amino-peptidase APN) which are responsible for the inactivation of the endogenous opioids; endorphins and enkephalins. Morphine and butorphanol exerts their analgesic effects by mimicking the actions of endorphins and enkephalins. The opiorphin analogue was synthesized based on the structure activity relationship of the amino acid sequence of opiorphin. The pharmacological profile of the analogue was tested by replacing Serine at position 4 with Proline. The hot plate and tail flick test were used to demonstrate the analgesic efficacy. There was a significant increase in the time for the tail flick response after an injection of opiorphin, which was similar to the morphine effect. There was no increase in time in the hot plate test after an injection of opiorphin. The results suggest that opiorphin works at spinal level only rather than both spinal and supraspinal. Further work is required to confirm our results. We did not find analgesic activity of the opiorphin analogue. Thus, Serine at position 4 is also important for its pharmacological action. Further work is required to illustrate the role of serine at position 4 in opiorphin.

Keywords: analgesic peptides, endogenous opioids, morphine, opiorphin

Procedia PDF Downloads 325

Authors: P. Rahnemoon, M. Sarabi Jamab, M. Javanmard Dakheli, A. Bostan

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In recent years, tendency to use of natural antimicrobial agents in food industry has increased. Pomegranate peels containing phenolic compounds and anti-microbial agents, are counted as valuable source for extraction of these compounds. In this study, the extraction of pomegranate peel extract was carried out at different ethanol/water ratios (40:60, 60:40, and 80:20), temperatures (25, 40, and 55 ˚C), and time durations (20, 24, and 28 h). The extraction yield, phenolic compounds, flavonoids, and anthocyanins were measured. ‎Antimicrobial activity of pomegranate peel extracts were determined against some food-borne ‎microorganisms such as Salmonella enteritidis, Escherichia coli, Listeria monocytogenes, ‎‎Staphylococcus aureus, Aspergillus niger, and Saccharomyces cerevisiae by agar diffusion and MIC methods. Results showed that at ethanol/water ratio 60:40, 25 ˚C and 24 h maximum amount of phenolic compounds ‎‏(‎‏‎349.518‎‏ ‏mg gallic acid‏/‏g dried extract), ‎flavonoids (250.124 mg rutin‏/‏g dried extract), anthocyanins (252.047 ‎‏‏mg ‎cyanidin‏‎3‎‏glucoside‏/‏‎100 g dried extract), and the strongest antimicrobial activity were obtained. ‎All extracts’ antimicrobial activities were demonstrated against every tested ‎‎microorganisms‏.‎‏ Staphylococcus aureus showed the highest sensitivity among the tested ‎‎‎microorganisms.

Keywords: antimicrobial agents, phenolic compounds, pomegranate peel, solvent extraction‎

Procedia PDF Downloads 258

Authors: Dmitriy Babenko, Sergey Yegorov, Ilya Korshukov, Aidana Sultanbekova, Valentina Barkhanskaya, Tatiana Bashirova, Yerzhan Zhunusov, Yevgeniya Li, Viktoriya Parakhina, Svetlana Kolesnichenko, Yeldar Baiken, Aruzhan Pralieva, Zhibek Zhumadilova, Matthew S. Miller, Gonzalo H. Hortelano, Anar Turmuhambetova, Antonella E. Chesca, Irina Kadyrova

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Introduction: The rapidly evolving COVID-19 pandemic, along with the re-emergence of pathogens causing acute respiratory infections (ARI), has necessitated the development of novel diagnostic tools to differentiate various causes of ARI. MALDI-MS, due to its wide usage and affordability, has been proposed as a potential instrument for diagnosing SARS-CoV-2 versus non-SARS-CoV-2 ARI. The aim of this study was to investigate the potential of MALDI-MS in conjunction with a machine learning model to accurately distinguish between symptomatic infections caused by SARS-CoV-2 and non-SARS-CoV-2 during both the early and later phases of the pandemic. Furthermore, this study aimed to analyze mass spectrometry (MS) data obtained from nasal swabs of healthy individuals. Methods: We gathered mass spectra from 252 samples, comprising 108 SARS-CoV-2-positive samples obtained in 2020 (Covid 2020), 7 SARS-CoV- 2-positive samples obtained in 2023 (Covid 2023), 71 samples from symptomatic individuals without SARS-CoV-2 (Control non-Covid ARVI), and 66 samples from healthy individuals (Control healthy). All the samples were subjected to RT-PCR testing. For data analysis, we employed the caret R package to train and test seven machine-learning algorithms: C5.0, KNN, NB, RF, SVM-L, SVM-R, and XGBoost. We conducted a training process using a five-fold (outer) nested repeated (five times) ten-fold (inner) cross-validation with a randomized stratified splitting approach. Results: In this study, we utilized the Covid 2020 dataset as a case group and the non-Covid ARVI dataset as a control group to train and test various machine learning (ML) models. Among these models, XGBoost and SVM-R demonstrated the highest performance, with accuracy values of 0.97 [0.93, 0.97] and 0.95 [0.95; 0.97], specificity values of 0.86 [0.71; 0.93] and 0.86 [0.79; 0.87], and sensitivity values of 0.984 [0.984; 1.000] and 1.000 [0.968; 1.000], respectively. When examining the Covid 2023 dataset, the Naive Bayes model achieved the highest classification accuracy of 43%, while XGBoost and SVM-R achieved accuracies of 14%. For the healthy control dataset, the accuracy of the models ranged from 0.27 [0.24; 0.32] for k-nearest neighbors to 0.44 [0.41; 0.45] for the Support Vector Machine with a radial basis function kernel. Conclusion: Therefore, ML models trained on MALDI MS of nasopharyngeal swabs obtained from patients with Covid during the initial phase of the pandemic, as well as symptomatic non-Covid individuals, showed excellent classification performance, which aligns with the results of previous studies. However, when applied to swabs from healthy individuals and a limited sample of patients with Covid in the late phase of the pandemic, ML models exhibited lower classification accuracy.

Keywords: SARS-CoV-2, MALDI-TOF MS, ML models, nasopharyngeal swabs, classification

Procedia PDF Downloads 108

Authors: Laidi Babouri

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The remarkable properties of steel, such as hardness and impact resistance, motivate their use in the automotive manufacturing industry. However, due to the very vulnerable environmental conditions of use, the steel that makes up the car body can corrode. This situation is motivating more and more automobile manufacturers to develop research to develop processes minimizing the rate of degradation of the physicomechanical properties of these materials. The present work falls within this perspective; it presents the results of a research study focused on the use of synthetic wax for the protection of steel, type XES (DC04), against corrosion in aggressive environments. The media used in this study are an acid medium with a pH=5.6, a 3% chloride medium, and a dry medium. Evaluation of the protective power of synthetic wax in different environments was carried out using mass loss techniques (immersion), completed by electrochemical techniques (stationary and transient). The results of the immersion of the steel samples, with a surface area of (1.44 cm²), in the various media, for a period of 30 days, using the immersion technique, showed high protective efficiency of synthetic wax in acidic and saline environments, with a lesser degree in a dry environment. Moreover, the study of the protective power, using electrochemical techniques, confirmed the results obtained in static mode (loss of mass), the protective efficiency of synthetic wax, against the corrosion of steel, in different environments, which reaches a maximum rate of 99.87% in a saline environment.

Keywords: corrosion, steel, industrial wax, environment, mass loss, electrochemical techniques

Procedia PDF Downloads 75

Authors: Nneka N. Uchegbu, Charles N. Ishiwu

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The nutritional composition and hypoglycaemic effect of crackers produced from a blend of sprouted pigeon pea, unripe plantain, and brewers’ spent grain and fed to Alloxan induced diabetic rat was investigated. Crackers were produced from different blends of sprouted pigeon pea, unripe plantain and brewers’ spent grain. The crackers were evaluated for proximate composition, amino acid profile and antinutritional factors. Blood glucose levels of normal and diabetic rats fed with the control sample and different formulations of cracker were measured. The protein content of the samples were significantly different (p < 0.05) from each other with sample A having the lowest value and sample B with the highest value. The values obtained showed that the samples contained most of the amino acids that are found in plant proteins. The levels of antinutritional factor determined were generally low. Administration of the formulated cracker meals led to a significant reduction in the fasting blood glucose level in the diabetic rats. The present study concluded that consumption of crackers produced from this composite flour can be recommended for the diabetics and those who are sceptical about the disease.

Keywords: crackers, diabetics rat, sprouted pigeon pea, unripe plantain and brewers’ spent grain

Procedia PDF Downloads 439

Authors: Raphael Odoh, Obida J. Oko, Mary S. Dauda

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The determination of Cd, Cr, Cu, Fe,Mn, Ni, Pb and Zn contents in edible oils (palm oil, ground-nut oil and soybean oil) bought from various markets of Benue and Taraba state were carried out with flame atomic absorption spectrophotometric technique. The method 3031 developed acid digestion of oils for metal analysis by atomic absorption or ICP spectrometry was used in the preparation of the edible oil samples for the determination of total metal content in this study. The overall results (µg/g) in palm oil sample ranged from 0.028-0.076, 0.035-0.092, 1.011-1.955, 2.101-4.892, 0.666-0.922, 0.054-0.095, 0.031-0.068 and 1.987-2.971 for Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn respectively, while in ground-nut oil the overall results ranged from 0.011-0.042, 0.011-0.052, 0.133-0.788, 1.789-2.511, 0.078-0.765, 0.045-0.092, 0.011-0.028 and 1.098-1.997 for Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn respectively. Of the heavy metals considered Cd and Ni showed the highest contamination in the soybean oil sample. The overall results in soybean oil samples ranged from 0.011-0.015, 0.017-0.032, 0.453-0.987, 1.789-2.511, 0.089-0.321, 0.011-0.016, 0.012-0.065 and 1.011-1.997 for Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn respectively. The concentration of Pb was the highest. The degree of contamination by each metal was estimated by the transfer factor. The transfer factors obtained for Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn in edible oils (palm oil, ground-nut oil and soybean oil) were 10.800, 16.500, 16.000, 18.813, 15.115, 14.230, 23.000 and 9.418 for Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn in palm oil, and 7.000, 12.500, 8.880, 11.333, 7.708, 10.833, 15.00 and 6.608 for Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn in ground-nut oil while for soybean oil the transfer factors were 13.000, 11.000, 7.642, 11.578, 4.486, 13.00, 12.333 and 4.412 for Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn respectively. The inter-element correlation was found among metals in edible oil samples using Pearson’s correlation co-efficient. There were positive and negative correlations among the metals determined. All Metals determined showed degree of contamination but concentrations lower than the USP specification.

Keywords: Benue State, contamination, edible oils, heavy metals, markets, Taraba State

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Authors: O. Smara, H. Dendougui, B. Legseir

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Atriplex halimus L. is particularly well adapted to arid and salt-affected areas. In this species, salinity resistance is often attributed to the presence of vesiculated hairs covering leaf surface and containing a large amount of salt. Atriplex halimus L. (Chenopodiaceae) is a perennial shrub native to the Mediterranean basin with excellent tolerance to drought and salinity. The species is present in semiarid to subhumid areas of the north Mediterranean and in arid zones from North Africa and the eastern Mediterranean. The main aim of this study was to identify a medicinal plant used in the Ouargla (Est-southern Algeria) for the treatment of several human pathologies. This plant is an important source for livestock in nitrogenous matter, it is an effective and relatively inexpensive tool in the fight against erosion and desertification and rehabilitation of degraded lands. Phytochemical investigation is applied to the majority of extracts of the powder of the aerial parts of Atriplex halimus L. Different chromatographic methods after liquid-liquid extraction are used; it is the thin layer chromatography (TLC) and paper using multiple systems and chemical revelations. This study followed by an evaluation by the phenol assay the Folin-Ciocalteu method, using gallic acid as a reference for phenols and quercetin for flavonols. Some polar extracts showed an interesting result better than the less polar extracts.

Keywords: Atriples halimus L., chenopodiaceae, flavonoids, phenols

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Authors: Madalin Enache, Simona Neagu, Roxana Cojoc, Ioana Gomoiu, Delia Ionela Dobre, Ancuta Roxana Trifoi

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Various types of halophilic and halotolerant microorganisms able to be cultivated in laboratory on culture media with a wide range of sodium chloride content are isolated from several salted environments. The extracellular enzymes of these microorganisms showed the enzymatic activity in these spectrums of salinity thus being attractive for several biotechnological processes developed at high ionic strength. In present work, a number of amylase, protease, esterase, lipase, cellulase, pectinase, xilanases and innulinase were identified for more than 50th bacterial strains isolated from water samples and sapropelic mud from four saline and hypersaline lakes located in Romanian plain. On the other hand, the cellulase and pectinase activity were also detected in some halotolerant microorganisms isolated from secondary agricultural flow of grapes processing. The preliminary data revealed that from totally tested strains seven harbor proteases activity, eight amylase activity, four for esterase and another four for lipase, three for pectinase and for one strain were identified either cellulase or pectinase activity. There were no identified enzymes able to hydrolase innulin added to culture media. Several strains isolated from sapropelic mud showed multiple extracellular enzymatic activities, namely three strains harbor three activities and another seven harbor two activities. The data revealed that amylase and protease activities were frequently detected if compare with other tested enzymes. In the case of pectinase were investigated, their ability to be used for increasing resveratrol recovery from material resulted after grapes processing. In this way, the resulted material from grapes processing was treated with microbial supernatant for several times (two, four and 24 hours) and the content of resveratrol was detected by High Performance Liquid Chromatography method (HPLC). The preliminary data revealed some positive results of this treatment.

Keywords: halophilic microorganisms, enzymes, pectinase, salinity

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Authors: Jawad Alzeer

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Natural products are well recognized as sources of drugs in several human ailments. To investigate the impact of variable extraction techniques on the cytotoxic effects of medicinal plant extracts, 5 well-known medicinal plants from Palestine were extracted with 90% ethanol, 80% methanol, acetone, coconut water, apple vinegar, grape vinegar or 5% acetic acid. The resulting extracts were screened for cytotoxic activities against three different cancer cell lines (B16F10, MCF-7, and HeLa) using a standard resazurin-based cytotoxicity assay and Nile Blue A as the positive control. Highly variable toxicities and tissue sensitivity were observed, depending upon the solvent used for extraction. Acetone consistently gave lower extraction yields but higher cytotoxicity, whereas other solvent systems gave much higher extraction yields with lower cytotoxicity. Interestingly, coconut water was found to offer a potential alternative to classical organic solvents; it gave consistently highest extraction yields, and in the case of S. officinalis L., highly toxic extracts towards MCF-7 cells derived from human breast cancer. These results demonstrate how the cytotoxicity of plant extracts can be inversely proportional to the yield, and that solvent selection plays an important role in both factors.

Keywords: plant extract, natural products, anti cancer drug, cytotoxicity

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Authors: Reza Vakili

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This experiment was conducted to investigate the effect of duration water restriction of drinking water and salinity level on production performance, egg quality and biochemical and hematological blood indices of laying hens. A total of 240 Hy-Line laying hens were used in a completely randomized design with a 2 × 2 factorial arrangement of treatments. Experimental treatments were: 1) free access to drinking water and a low level of salinity (TDS below 500 mg/L) (FAW+LS), 2) free access to water and a high level of salinity (TDS above 1500 mg/L), (FAW+HS), 3) 12 h nightly water restriction and a low level of salinity (LAW+LS), and 4) 12 h water restriction and a high level of salinity (LAW+HS). Intake of feed, percentage of egg production and egg weight and mass were not affected by water restriction or salinity level (P > 0.05), however, a trend (P < 0.01) for lower water consumption was detected in water-restricted hens, regardless of salinity level (213 vs 187). A tendency for lower eggshell and yolk weights was observed in hens that had limited access to water with high salinity compared to those had free access to high saline water (P = 0.08). Serum total protein and glucose concentrations significantly reduced (P < 0.05) in hens drank high salinity water, regardless of water restriction. Moreover, saline water increased the concentration of uric acid, creatinine, and cholesterol when compared to low salinity drank-hens (P < 0.05). The concentrations of ALT and AST increased with salinity level (P < 0.05) and water restriction caused an increment in AST content (P < 0.05). In conclusion, Hy-Line laying hens could withstand water restriction, whilst could not tolerate water salinity of about 1500 mg/L.

Keywords: chemical pollutants, eggs, laying hens, salinity, water quality

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Authors: Lynn Lehmann, Dinorah Leyva, Ana Lazic, Stefan Duhr, Philipp Baaske

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Characterization of biomolecular interactions, such as protein-protein, protein-nucleic acid or protein-small molecule, provides critical insights into cellular processes and is essential for the development of drug diagnostics and therapeutics. Here we present a novel, label-free, and tether-free technology to analyze picomolar to millimolar affinities of biomolecular interactions by Microscale Thermophoresis (MST). The entropy of the hydration shell surrounding molecules determines thermophoretic movement. MST exploits this principle by measuring interactions using optically generated temperature gradients. MST detects changes in the size, charge and hydration shell of molecules and measures biomolecule interactions under close-to-native conditions: immobilization-free and in bioliquids of choice, including cell lysates and blood serum. Thus, MST measures interactions under close-to-native conditions, and without laborious sample purification. We demonstrate how MST determines the picomolar affinities of antibody::antigen interactions, and protein::protein interactions measured from directly from cell lysates. MST assays are highly adaptable to fit to the diverse requirements of different and complex biomolecules. NanoTemper´s unique technology is ideal for studies requiring flexibility and sensitivity at the experimental scale, making MST suitable for basic research investigations and pharmaceutical applications.

Keywords: biochemistry, biophysics, molecular interactions, quantitative techniques

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Authors: Said M. AL-Mashaikhi, El-Said I. El-Shafey, Fakhreldin O. Suliman, Saleh Al-Busafi

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Activated carbon (AC) was prepared from date palm leaflets via NaOH activation. AC was oxidized using nitric acid, producing oxidized activated carbon (OAC). OAC was surface functionalized using different amine surfactants, including methylamine (ONM), ethylamine (ONE), and diethylamine (ONDE) using the amide coupling process. Produced carbons were surface characterized for surface area and porosity, X-ray diffraction, SEM, FTIR, and TGA. AC surface area (580 m²/g) has shown a decrease in oxidation to 260 m²/g for OAC. On amine functionalization, the surface area has further decreased to 218, 108, and 20 m²/g on functionalization with methylamine, ethylamine, and diethylamine, respectively. FTIR and TGA showed that the nature of amine functionalization of AC is chemical. Methylene blue sorption was tested on these carbons in terms of kinetics and equilibrium. Sorption was found faster on amine-functionalized carbons than both AC and OAC, and this is due to hydrophobic interaction with the alkyl groups immobilized with data following pseudo second-order reaction. On the other hand, AC showed the slowest adsorption kinetic process due to the diffusion in the porous structure of AC. Sorption equilibrium data was found to follow the Langmuir sorption isotherm with maximum sorption found on ONE. Regardless of its lower surface area than activated carbon, ethylamine functionalized AC showed better performance than AC in terms of kinetics and equilibrium for dye removal.

Keywords: activated carbon, dye removal, functionalization, hydrophobic interaction, water treatment

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Authors: Anamarel Medina-Hernandez, Teresa Ponce-Noyola, Ileana Vera-Reyes, Ana C. Ramos-Valdivia

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Somatic embryos reproduce original seed characteristics and could be implemented in biotechnological studies. Jatropha curcas L. is an important plant for biodiesel production, but also is used in traditional medicine. Seeds from J. curcas are toxic because contain diterpenoids called phorbol esters, but in Mexico exist a non-toxic variety. Therefore, somatic embryos suspension cultures from non-toxic J. curcas variety were induced. In order to investigate the characteristics of somatic embryos, a differential proteomic analysis was made between pre-globular and globular stages by 2-D gel electrophoresis. 108 spots were differentially expressed (p<0.02), and 20 spots from globular somatic embryos were sequenced by MALDI-TOF-TOF mass spectrometry. A comparative analysis of terpenoids production between the two stages was made by RP-18 TLC plates. The sequenced proteins were related to energy production (68%), protein destination and storage (9%), secondary metabolism (9%), signal transduction (5%), cell structure (5%) and aminoacid metabolism (4%). Regarding terpenoid production, in pre-globular and globular somatic embryos were identified sterols and triterpenes of pharmacological interest (alpha-amyrin and betulinic acid) but also it was found compounds that were unique to each stage. The results of this work are the basis to characterize at different levels the J. curcas somatic embryos so that this system can be used efficiently in biotechnological processes.

Keywords: Jatropha curcas, proteomics, somatic embryo, terpenoids

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Authors: Lai Peng, Cristina Pintucci, Dries Seuntjens, José Carvajal-Arroyo, Siegfried Vlaeminck

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Economic incentives drive the implementation of short-cut nitrogen removal processes such as nitritation/denitritation (Nit/DNit) to manage nitrogen in waste streams devoid of biodegradable organic carbon. However, as any biological nitrogen removal process, the potent greenhouse gas nitrous oxide (N2O) could be emitted from Nit/DNit. Challenges remain in understanding the fundamental mechanisms and development of engineered mitigation strategies for N2O production. To provide answers, this work focuses on manure as a model, the biggest wasted nitrogen mass flow through our economies. A sequencing batch reactor (SBR; 4.5 L) was used treating the centrate (centrifuge supernatant; 2.0 ± 0.11 g N/L of ammonium) from an anaerobic digester processing mainly pig manure, supplemented with a co-substrate. Glycerin was used as external carbon source, a by-product of vegetable oil. Out-selection of nitrite oxidizing bacteria (NOB) was targeted using a combination of low dissolved oxygen (DO) levels (down to 0.5 mg O2/L), high temperature (35ºC) and relatively high free ammonia (FA) (initially 10 mg NH3-N/L). After reaching steady state, the process was able to remove 100% of ammonium with minimum nitrite and nitrate in the effluent, at a reasonably high nitrogen loading rate (0.4 g N/L/d). Substantial N2O emissions (over 15% of the nitrogen loading) were observed at the baseline operational condition, which were even increased under nitrite accumulation and a low organic carbon to nitrogen ratio. Yet, higher DO (~2.2 mg O2/L) lowered aerobic N2O emissions and weakened the dependency of N2O on nitrite concentration, suggesting a shift of N2O production pathway at elevated DO levels. Limiting the greenhouse gas emissions (environmental protection) from such a system could be substantially minimized by increasing the external carbon dosage (a cost factor), but also through the implementation of an intermittent aeration and feeding strategy. Promising steps forward have been presented in this abstract, yet at the conference the insights of ongoing experiments will also be shared.

Keywords: mitigation, nitrous oxide, nitritation/denitritation, pig manure

Procedia PDF Downloads 249

Authors: Bouchiha Hanene, Rouabhi Rachid, Bouchama Khaled, Djebar Berrebbah Houraya, Djebar Mohamed Reda

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Recently, some natural products such as essentials oils (EOs) have been used in the fields as alternative to synthetic compounds, to minimize the negative impacts to the environment. This fact has led to questions about the possible impact of EOs on ecosystems. Currently in toxicology, the use of alternative models can help to understand the mechanisms of toxic action, at different levels of organization of ecosystems. Algae, protozoa and bacteria form the base of the food chain and protozoan cells are used as bioindicators often of pollution in environment. Unicellular organisms offer the possibility of direct study of independent cells with specific characteristics of individual cells and whole organisms at the same time. This unicellular facilitates the study of physiological processes, and effects of pollutants at the cellular level, which makes it widely used to assess the toxic effects of various xenobiotics. This study aimed to verify the effects of EOs of one famous plant used tremendously in our folk medicine, namely Artemisia herba alba in causing acute toxicity (24 hours) and chronic (15 days) toxicity for model cellular (Paramecium sp). To this end, cellular’s of paramecium were exposed to various concentrations (Three doses were chosen) of EOs extracted from plant (Artemisia herba alba). In the first experiment, the cellular s cultures were exposed for 48 hours to different concentrations to determine the median lethal concentration (DL50). We followed the evolution of physiological parameters (growth), biochemical (total proteins, respiratory metabolism), as well as the variations of a bio marker the GSH. Our results highlighted a light inhibition of the growth of the protozoa as well as a disturbance of the contents of total proteins and a reduction in the reduced rate of glutathione. The polarographic study revealed a stimulation of the consumption of O2 and this at the treated cells.

Keywords: essential oils, protozoa, bio indicators, toxicity, Growth, bio marker, proteins, polarographic

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Authors: John Abraham, Ramesh Saravana Kumar, Francis, Xavier, Deepak Mathew

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Broiler slaughter waste has become a major source of pollution throughout the world. Utilization of broiler slaughter waste by dry rendering process produced Rendered Chicken Oil (RCO) a cheap raw material for biodiesel production and Carcass Meal a feed ingredient for pets and fishes. Conversion of RCO into biodiesel may open new vistas for generating wealth from waste besides controlling the major havoc of environmental pollution. A two-step process to convert RCO to good quality Biodiesel was invented. Acid catalysed esterification of FFA followed by base catalysed transesterification of triglycerides was carried out after meticulously standardising the methanol molar ratio, catalyst concentration, reaction temperature and reaction time to obtain the maximum biodiesel yield of 97.62% and lowest glycerol yield of 6.96%. RCO biodiesel blended was tested in a Mahindra Scorpio CRDI engine. The results revealed that the blending of commercial diesel with 20% RCO biodiesel lead to less engine wear, a quieter engine and better fuel economy. The better lubricating qualities of RCO B20 prevented over heating of engine, which prolongs the engine life. The blending of biodiesel at 20% to commercial diesel can reduce the import of costly crude oil and simultaneously, substantially reduce the engine emissions as proved by significantly lower smoke levels, thus mitigating climatic changes.

Keywords: broiler waste, rendered chicken oil, biodiesel, engine testing

Procedia PDF Downloads 435

Authors: Amina Aloui, Jalila Ben Salah-Abbès, Abdellah Zinedine, Amar Riba, Noel Durand, Catherine Brabet, Didier Montet, Samir Abbès

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Mycotoxin contamination of food and feed-isa globaconcern, both economically and for public health. Aflatoxin M1 (AFM1) is the principal hydroxylated metabolite of aflatoxin B1. It is frequently found in milk and other dairy products. It is responsible for the development of hepatocellular carcinoma and immunotoxic in humans and animals. The reduction of its bioavailabilitybecomesa great demand in order to protect human and animal health. The use of probiotic bacteria and clay are demonstrated to be able to bind AFM1 in vitro. This study aimed to investigate, in vivo, the activity of two-component mixture: L. rhamnosusGAF06 (LR) and bentonite for reducing the oxidative stress and the histological alterationsinduced by AFM1 in the liver andkidneys. For the experiment, male mice were divided into 7 groups (6 mice/group) and treated, orally, by AFM1, alone or in combination with LR and/or bentonite, for 10 days as follows: group 1 control, group 2 treated with LR alone (2.108 CFU/ml), group 3 treated with bentonite alone (1g/kg), group 4 treated with AFM1 alone (100μg/kg), group 5 co-treated with LR+AFM1, group 6 co-treated with bentonite+AFM1, group 7 co-treated with bentonite+LR+AFM1. At the end of the treatment, the mice were sacrificed, and the livers and kidneys were collected for histological assays. Intracellular antioxidant activities and lipid peroxidation were also studied. The results showed that AFM1causeddamage in liver and kidney tissues, being evidence of hepatotoxicity and nephrotoxicity marked by necrotic cells. It increased the MDA level and decreased the antioxidant enzyme activities (SOD) in both organs. In contrast, the co-treatment with AFM1 plus LR and/or bentonitesignificantly improved the hepatic and renal tissues, regulated kidney, and liver antioxidant enzyme activities. This improvement was more remarkable with the administration of LR-bentonite mixture with AFM1.LR and bentonite alone showed to be safe during the treatment. This mixture can be a promising candidate for future applications in biotechnological processes that aimed to detoxify AFM1in food and feed.

Keywords: aflatoxin M1, bentonite, L. rhamnosus GAF06, oxidative stress, prevention

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Authors: Vanessa De Carvalho, Chad MacPherson, Julien Tremblay, Julie Champagne, Stephanie-Anne Girard

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Background: The interactions between bacteria and fungi in the human vaginal microbiome are fundamental to the concept of health and disease. The means by which the microbiota and mycobiota interact is still poorly understood and further studies are necessary to properly characterize this complex ecosystem. The aim of this study was to select a DNA extraction method capable of recovering high qualities of fungal and bacterial DNA from a single vaginal swab. Methods: 11 female volunteers ( ≥ 20 to < 55 years old) self-collected vaginal swabs in triplicates. Three commercial extraction kits: Masterpure Yeast Purification kit (Epicenter), PureLink™ Microbiome DNA Purification kit (Invitrogen), and Quick-DNA™ Fecal/Soil Microbe Miniprep kit (Zymo) were evaluated on the ability to recover fungal and bacterial DNA simultaneously. The extraction kits were compared on the basis of recovery, yield, purity, and the community richness of bacterial (16S rRNA - V3-V4 region) and fungal (ITS1) microbiota composition by Illumina MiSeq amplicon sequencing. Results: Recovery of bacterial DNA was achieved with all three kits while fungal DNA was only consistently recovered with Masterpure Yeast Purification kit (yield and purity). Overall, all kits displayed similar microbiota profiles for the top 20 OTUs; however, Quick-DNA™ Fecal/Soil Microbe Miniprep kit (Zymo) showed more species richness than the other two kits. Conclusion: In the present study, Masterpure Yeast purification kit proved to be a good candidate for purification of high quality fungal and bacterial DNA simultaneously. These findings have potential benefits that could be applied in future vaginal microbiome research. Whilst the use of a single extraction method would lessen the burden of multiple swab sampling, decrease laboratory workload and off-set costs associated with multiple DNA extractions, thoughtful consideration must be taken when selecting an extraction kit depending on the desired downstream application.

Keywords: bacterial vaginosis, DNA extraction, microbiota, mycobiota, vagina, vulvovaginal candidiasis, women’s health

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Authors: Hee-Jae Suh, Mi-Na Hong, Min-Ji Kim, Yeon-Seong Jeong, Ok-Hwan Lee, Jae-Wook Shin, Hyang-Sook Chun, Chan Lee

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Brown HT is a bis-azo dye which is permitted in EU as a food colorant. So far, many studies have focused on HPLC using diode array detection (DAD) analysis for detection of this food colorant with different columns and mobile phases. Even though these methods make it possible to detect Brown HT, low recovery, reproducibility, and linearity are still the major limitations for the application in foods. The purpose of this study was to compare various methods for the analysis of Brown HT and to develop an improved analytical methods including pretreatment. Among tested analysis methods, best resolution of Brown HT was observed when the following solvent was applied as a eluent; solvent A of mobile phase was 0.575g NH4H2PO4, and 0.7g Na2HPO4 in 500mL water added with 500mL methanol. The pH was adjusted using phosphoric acid to pH 6.9 and solvent B was methanol. Major peak for Brown HT appeared at the end of separation, 13.4min after injection. This method exhibited relatively high recovery and reproducibility compared with other methods. LOD (0.284 ppm), LOQ (0.861 ppm), resolution (6.143), and selectivity (1.3) of this method were better than those of ammonium acetate solution method which was most frequently used. Precision and accuracy were verified through inter-day test and intra-day test. Various methods for sample pretreatments were developed for different foods and relatively high recovery over 80% was observed in all case. This method exhibited high resolution and reproducibility of Brown HT compared with other previously reported official methods from FSA and, EU regulation.

Keywords: analytic method, Brown HT, food colorants, pretreatment method

Procedia PDF Downloads 478