Search results for: laboratory investigations

Authors: Elisa Esposito, Johannes C. Jansen, Loredana Dellamuzia, Ugo Moretti, Lidietta Giorno

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The reduction of CO₂ emission into the atmosphere as a result of human activity is one of the most important environmental challenges to face in the next decennia. Emission of CO₂, related to the use of fossil fuels, is believed to be one of the main causes of global warming and climate change. In this scenario, the production of biomethane from organic waste, as a renewable energy source, is one of the most promising strategies to reduce fossil fuel consumption and greenhouse gas emission. Unfortunately, biogas upgrading still produces the greenhouse gas CO₂ as a waste product. Therefore, this work presents a case study on biogas upgrading, aimed at the simultaneous purification of methane and CO₂ via different steps, including CO₂/methane separation by polymeric membranes. The original objective of the project was the biogas upgrading to distribution grid quality methane, but the innovative aspect of this case study is the further purification of the captured CO₂, transforming it from a useless by-product to a pure gas with food-grade quality, suitable for commercial application in the food and beverage industry. The study was performed on a pilot plant constructed by Tecno Project Industriale Srl (TPI) Italy. This is a model of one of the largest biogas production and purification plants. The full-scale anaerobic digestion plant (Montello Spa, North Italy), has a digestive capacity of 400.000 ton of biomass/year and can treat 6.250 m3/hour of biogas from FORSU (organic fraction of solid urban waste). The entire upgrading process consists of a number of purifications steps: 1. Dehydration of the raw biogas by condensation. 2. Removal of trace impurities such as H₂S via absorption. 3.Separation of CO₂ and methane via a membrane separation process. 4. Removal of trace impurities from CO₂. The gas separation with polymeric membranes guarantees complete simultaneous removal of microorganisms. The chemical purity of the different process streams was analysed by a certified laboratory and was compared with the guidelines of the European Industrial Gases Association and the International Society of Beverage Technologists (EIGA/ISBT) for CO₂ used in the food industry. The microbiological purity was compared with the limit values defined in the European Collaborative Action. With a purity of 96-99 vol%, the purified methane respects the legal requirements for the household network. At the same time, the CO₂ reaches a purity of > 98.1% before, and 99.9% after the final distillation process. According to the EIGA/ISBT guidelines, the CO₂ proves to be chemically and microbiologically sufficiently pure to be suitable for food-grade applications.

Keywords: biogas, CO₂ separation, CO2 utilization, CO₂ food grade

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Authors: E. Nemes-Nagy, E. Fogarasi, M. Croitoru, A. Nyárádi, K. Komlódi, S. Pál, A. Kovács, O. Kopácsy, R. Tripon, Z. Fazakas, C. Uzun, Z. Simon-Szabó, V. Balogh-Sămărghițan, E. Ernő Nagy, M. Szabó, M. Tilinca

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Intense oxidative stress, increased glycated hemoglobin and mineral imbalance represent risk factors for complications in diabetic patients. Cardiovascular complications are most common in these patients, including nephropathy. This study was conducted in 2015 at the Procardia Laboratory in Tîrgu Mureș, Romania on 40 type 2 diabetic adults. Routine biochemical tests were performed on the Konleab 20XTi analyzer (serum glucose, total cholesterol, LDL and HDL cholesterol, triglyceride, creatinine, urea). We also measured serum uric acid, magnesium and calcium concentration by photometric procedures, potassium, sodium and chloride by ion selective electrode, and chromium by atomic absorption spectrometry in a group of patients. Glycated hemoglobin (HbA1c) dosage was made by reflectometry. Urine analysis was performed using the HandUReader equipment. The level of oxidative stress was measured by serum malondialdehyde dosage using the thiobarbituric acid reactive substances method. MDRD (Modification of Diet in Renal Disease) formula was applied for calculation of creatinine-derived glomerular filtration rate. GraphPad InStat software was used for statistical analysis of the data. The diabetic subject included in the study presented high MDA concentrations, showing intense oxidative stress. Calcium was deficient in 5% of the patients, chromium deficiency was present in 28%. The atherogenic cholesterol fraction was elevated in 13% of the patients. Positive correlation was found between creatinine and MDRD-creatinine values (p<0.0001), 68% of the patients presented increased creatinine values. The majority of the diabetic patients had good control of their diabetes, having optimal HbA1c values, 35% of them presented fasting serum glucose over 120 mg/dl and 18% had glucosuria. Intense oxidative stress and mineral deficiencies can increase the risk of cardiovascular complications in diabetic patients in spite of their good metabolic balance. More than two third of the patients present biochemical signs of nephropathy, cystatin C dosage and microalbuminuria could reveal better the kidney disorder, but glomerular filtration rate calculation formulas are also useful for evaluation of renal function.

Keywords: cardiovascular risk, homocysteine, malondialdehyde, metformin, minerals, type 2 diabetes, vitamin B12

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Authors: Karthick Dharmalingam, Stalin Ramakrishnan, Haseena Banu Hedayathullah Khan, Sachidanandanam Thiruvaiyaru Panchanadham, Shanthi Palanivelu

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Background: Breast cancer is arising as the most dreadful cancer affecting women worldwide. Hence, there arises a need to search and test for new drugs. Herbal formulations used in Siddha preparations are proved to be effective against various types of cancer. They also offer advantage through synergistic amplification and diminish any possible adverse effects. Tridham (TD) is a herbal formulation prepared in our laboratory consisting of Terminalia chebula, Elaeocarpus ganitrus and Prosopis cineraria in a definite ratio and has been used for the treatment of mammary carcinoma. Objective: To study the restorative effect of Tridham and penta galloyl glucose (a component of TD) on DMBA induced mammary carcinoma in female Sprague Dawley rats. Materials and Methods: Rats were divided into seven groups of six animals each. Group I (Control) received corn oil. Group II– mammary carcinoma was induced by DMBA dissolved in corn oil single dose orally. Group III and Group IV were induced with DMBA and subsequently treated with Tridham and penta galloyl glucose, respectively for 48 days. Group V was treated with DMBA and subsequently with a standard drug, cyclophosphamide. Group VI and Group VII were given Tridham and penta galloyl glucose alone, respectively for 48 days. After the experimental period, the animals were sacrificed by cervical decapitation. The mammary gland tissue was excised and levels of antioxidants were determined by biochemical assay. p53 and PCNA expression were accessed using immunohistochemistry. Nrf-2, Cox-2 and caspase-3 protein expression were studied by Western Blotting analysis. p21, Bcl-2, Bax, Bad and caspase-8 gene expression were studied by RT-PCR. Results: Histopathological studies confirmed induction of mammary carcinoma in DMBA induced rats and treatment with TD and PGG resulted in regression of tumour. The levels of enzymic and non-enzymic antioxidants were decreased in DMBA induced rats when compared to control rats. The levels of cell cycle inhibitory markers and apoptotic markers were decreased in DMBA induced rats when compared to control rats. These parameters were restored to near normal levels on treatment with Tridham and PGG. Conclusion: The results of the present study indicate the antineoplastic effect of Tridham and PGG are exerted through the modulation of antioxidant status and expression of cell cycle regulatory markers as well as apoptotic markers. Acknowledgment: Financial assistance provided in the form of ICMR-SRF by Indian Council of Medical Research (ICMR), India is gratefully acknowledged here.

Keywords: antioxidants, Mammary carcinoma, pentaGalloyl glucose, Tridham

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Authors: B. T. Naveen Kumar, Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, A. H. Shanthanagouda, Orawan Boodde, K. M. Shankar, Prakash Patil, Shubhkaramjeet Kaur

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Early mortality syndrome (EMS)/Acute Hepatopancreatic Necrosis Disease (AHPND) has emerged as a major obstacle for the shrimp farming around the world. It is caused by a strain of Vibrio parahaemolyticus. The possible preventive and control measure is, early and rapid detection of the pathogen in the broodstock, post-larvae and monitoring the shrimp during the culture period. Polymerase chain reaction (PCR) based early detection methods are good, but they are costly, time taking and requires a sophisticated laboratory. The present study was conducted to develop a simple, sensitive and rapid diagnostic farmer level kit for the reliable detection of AHPND in shrimp. A panel of monoclonal antibodies (MAbs) were raised against the recombinant Pir B protein (rPirB). First, an immunodot was developed by using MAbs G3B8 and Mab G3H2 which showed specific reactivity to purified r-PirB protein with no cross-reactivity to other shrimp bacterial pathogens (AHPND free Vibrio parahaemolyticus (Indian strains), V. anguillarum, WSSV, Aeromonas hydrophila, and Aphanomyces invadans). Immunodot developed using Mab G3B8 is more sensitive than that with the Mab G3H2. However, immunodot takes almost 2.5 hours to complete with several hands-on steps. Therefore, the flow-through assay (FTA) was developed by using a plastic cassette containing the nitrocellulose membrane with absorbing pads below. The sample was dotted in the test zone on the nitrocellulose membrane followed by continuos addition of five solutions in the order of i) blocking buffer (BSA) ii) primary antibody (MAb) iii) washing Solution iv) secondary antibody and v) chromogen substrate (TMB) clear purple dots against a white background were considered as positive reactions. The FTA developed using MAbG3B8 is more sensitive than that with MAb G3H2. In FTA the two MAbs showed specific reactivity to purified r-PirB protein and not to other shrimp bacterial pathogens. The FTA is simple to farmer/field level, sensitive and rapid requiring only 8-10 min for completion. Tests can be developed to kits, which will be ideal for use in biosecurity, for the first line of screening (at the port or pond site) and during monitoring and surveillance programmes overall for the good management practices to reduce the risk of the disease.

Keywords: acute hepatopancreatic necrosis disease, AHPND, flow-through assay, FTA, farmer level, immunodot, pond site, shrimp

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Authors: Shaher A. I. Shalfawi, Fredrik Hviding, Bjornar Kjellstadli

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The main purpose of this study was to measure the effect of two different strength training methods on muscle strength, muscle mass, fat mass and endurance factors. Fourteen physical education students accepted to participate in this study. The participants were then randomly divided into three groups, traditional training group (TTG), cluster training group (CTG) and control group (CG). TTG consisted of 4 participants aged ( ± SD) (22.3 ± 1.5 years), body mass (79.2 ± 15.4 kg) and height (178.3 ± 11.9 cm). CTG consisted of 5 participants aged (22.2 ± 3.5 years), body mass (81.0 ± 24.0 kg) and height (180.2 ± 12.3 cm). CG consisted of 5 participants aged (22 ± 2.8 years), body mass (77 ± 19 kg) and height (174 ± 6.7 cm). The participants underwent a hypertrophy strength training program twice a week consisting of 4 sets of 10 reps at 70% of one-repetition maximum (1RM), using barbell squat and barbell bench press for 8 weeks. The CTG performed 2 x 5 reps using 10 s recovery in between repetitions and 50 s recovery between sets, while TTG performed 4 sets of 10 reps with 90 s recovery in between sets. Pre- and post-tests were administrated to assess body composition (weight, muscle mass, and fat mass), 1RM (bench press and barbell squat) and a laboratory endurance test (Bruce Protocol). Instruments used to collect the data were Tanita BC-601 scale (Tanita, Illinois, USA), Woodway treadmill (Woodway, Wisconsin, USA) and Vyntus CPX breath-to-breath system (Jaeger, Hoechberg, Germany). Analysis was conducted at all measured variables including time to peak VO2, peak VO2, heart rate (HR) at peak VO2, respiratory exchange ratio (RER) at peak VO2, and number of breaths per minute. The results indicate an increase in 1RM performance after 8 weeks of training. The change in 1RM squat was for the TTG = 30 ± 3.8 kg, CTG = 28.6 ± 8.3 kg and CG = 10.3 ± 13.8 kg. Similarly, the change in 1RM bench press was for the TTG = 9.8 ± 2.8 kg, CTG = 7.4 ± 3.4 kg and CG = 4.4 ± 3.4 kg. The within-group analysis from the oxygen consumption measured during the incremental exercise indicated that the TTG had only a statistical significant increase in their RER from 1.16 ± 0.04 to 1.23 ± 0.05 (P < 0.05). The CTG had a statistical significant improvement in their HR at peak VO2 from 186 ± 24 to 191 ± 12 Beats Per Minute (P < 0.05) and their RER at peak VO2 from 1.11 ± 0.06 to 1.18 ±0.05 (P < 0.05). Finally, the CG had only a statistical significant increase in their RER at peak VO2 from 1.11 ± 0.07 to 1.21 ± 0.05 (P < 0.05). The between-group analysis showed no statistical differences between all groups in all the measured variables from the oxygen consumption test during the incremental exercise including changes in muscle mass, fat mass, and weight (kg). The results indicate a similar effect of hypertrophy strength training irrespective of the methods of the training used on untrained subjects. Because there were no notable changes in body-composition measures, the results suggest that the improvements in performance observed in all groups is most probably due to neuro-muscular adaptation to training.

Keywords: hypertrophy strength training, cluster set, Bruce protocol, peak VO2

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Authors: Kartik Gupta, Virendra Kumar, Sanjeev Sinha, N. Jagannathan

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Introduction: A significant number of patients having human immunodeficiency virus (HIV) infection show a neurocognitive decline (NCD) ranging from minor cognitive impairment to severe dementia. The possible causes of NCD in HIV-infected patients include brain injury by HIV before cART, neurotoxic viral proteins and metabolic abnormalities. In the present study, we compared the level of NCD in asymptomatic HIV-infected patients with changes in brain metabolites measured by using magnetic resonance spectroscopy (MRS). Methods: 43 HIV-positive patients (30 males and 13 females) coming to ART center of the hospital and HIV-seronegative healthy subjects were recruited for the study. All the participants completed MRI and MRS examination, detailed clinical assessments and a battery of neuropsychological tests. All the MR investigations were carried out at 3.0T MRI scanner (Ingenia/Achieva, Philips, Netherlands). MRI examination protocol included the acquisition of T2-weighted imaging in axial, coronal and sagittal planes, T1-weighted, FLAIR, and DWI images in the axial plane. Patients who showed any apparent lesion on MRI were excluded from the study. T2-weighted images in three orthogonal planes were used to localize the voxel in left frontal lobe white matter (FWM) and left basal ganglia (BG) for single voxel MRS. Single voxel MRS spectra were acquired with a point resolved spectroscopy (PRESS) localization pulse sequence at an echo time (TE) of 35 ms and a repetition time (TR) of 2000 ms with 64 or 128 scans. Automated preprocessing and determination of absolute concentrations of metabolites were estimated using LCModel by water scaling method and the Cramer-Rao lower bounds for all metabolites analyzed in the study were below 15\%. Levels of total N-acetyl aspartate (tNAA), total choline (tCho), glutamate + glutamine (Glx), total creatine (tCr), were measured. Cognition was tested using a battery of tests validated for Indian population. The cognitive domains tested were the memory, attention-information processing, abstraction-executive, simple and complex perceptual motor skills. Z-scores normalized according to age, sex and education standard were used to calculate dysfunction in these individual domains. The NCD was defined as dysfunction with Z-score ≤ 2 in at least two domains. One-way ANOVA was used to compare the difference in brain metabolites between the patients and healthy subjects. Results: NCD was found in 23 (53%) patients. There was no significant difference in age, CD4 count and viral load between the two groups. Maximum impairment was found in the domains of memory and simple motor skills i.e., 19/43 (44%). The prevalence of deficit in attention-information processing, complex perceptual motor skills and abstraction-executive function was 37%, 35%, 33% respectively. Subjects with NCD had a higher level of Glutamate in the Frontal region (8.03 ± 2.30 v/s. 10.26 ± 5.24, p-value 0.001). Conclusion: Among newly diagnosed, ART-naïve retroviral disease patients from India, cognitive decline was found in 53\% patients using tests validated for this population. Those with neurocognitive decline had a significantly higher level of Glutamate in the left frontal region. There was no significant difference in age, CD4 count and viral load at initiation of ART between the two groups.

Keywords: HIV, neurocognitive decline, neurometabolites, magnetic resonance spectroscopy

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Authors: Tomas Makaras

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Sticklebacks (Gasterosteiformes) are increasingly used in ecological and evolutionary research and become well-established role as model species for biologists. However, ecotoxicology studies concerning behavioural effects in sticklebacks regarding stress responses, mainly induced by chemical mixtures, have hardly been addressed. Moreover, although many authors in their studies emphasised the similarity between three-spined and nine-spined stickleback in morphological, neuroanatomical and behavioural adaptations to environmental changes, several comparative studies have revealed considerable differences between these species in and their susceptibility and resistance to variousstressors in laboratory experiments. The hypothesis of this study was that three-spined and nine-spined stickleback species will demonstrate apparent differences in response patterns and sensitivity to metal-based chemicals stimuli. For this purpose, we investigated the swimming behaviour (including mortality rate based on 96-h LC50 values) of two ecologically similar three-spined (Gasterosteusaculeatus) and nine-spined sticklebacks (Pungitiuspungitius) to short-term (up to 24 h) metal mixture (MIX) exposure. We evaluated the relevance and efficacy of behavioural responses of test species in the early toxicity assessment of chemical mixtures. Fish exposed to six (Zn, Pb, Cd, Cu, Ni and Cr) metals in the mixture were either singled out by the Water Framework Directive as priority or as relevant substances in surface water, which was prepared according to the environmental quality standards (EQSs) of these metals set for inland waters in the European Union (EU) (Directive 2013/39/EU). Based on acute toxicity results, G. aculeatus found to be slightly (1.4-fold) more tolerant of MIX impact than those of P. pungitius specimens. The performed behavioural analysis showed the main effect on the interaction between time, species and treatment variables. Although both species exposed to MIX revealed a decreasing tendency in swimming activity, these species’ responsiveness to MIX was somewhat different. Substantial changes in the activity of G. aculeatus were established after 3-h exposure to MIX solutions, which was 1.43-fold lower, while in the case of P. pungitius, 1.96-fold higher than established 96-h LC50 values for each species. This study demonstrated species-specific differences in response sensitivity to metal-based water pollution, indicating behavioural insensitivity of P. pungitiuscompared to G. aculeatus. While many studies highlight the usefulness and suitability of nine-spined sticklebacks for evolutionary and ecological research, attested by their increasing popularity in these fields, great caution must be exercised when using them as model species in ecotoxicological research to probe metal contamination. Meanwhile, G. aculeatus showed to be a promising bioindicator species in the environmental ecotoxicology field.

Keywords: acute toxicity, comparative behaviour, metal mixture, swimming activity

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Authors: Usha S. Adiga, Sachidanada Adiga

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Introduction: Diabetes Mellitus (DM) is one of the most common non-communicable diseases which imposes a large economic burden on the global health-care system. Cost of illness studies in India have assessed the health care cost of DM, but have certain limitations due to lack of standardization of the methods used, improper documentation of data, lack of follow up, etc. The objective of the study was to estimate the cost of illness of uncomplicated versus complicated type 2 diabetes mellitus in Coastal Karnataka, India. The study also aimed to find out the trend of cost of illness of the disease over a decade. Methodology: A prevalence based bottom-up approach study was carried out in two tertiary care hospitals located in Coastal Karnataka after ethical approval. Direct Medical costs like annual laboratory costs, pharmacy cost, consultation charges, hospital bed charges, surgical /intervention costs of 238 diabetics and 340 diabetic patients respectively from two hospitals were obtained from the medical record sections. Patients were divided into six groups, uncomplicated diabetes, diabetic retinopathy(DR), nephropathy(DN), neuropathy(DNeu), diabetic foot(DF), and ischemic heart disease (IHD). Different costs incurred in 2008 and 2017 in these groups were compared, to study the trend of cost of illness. Kruskal Wallis test followed by Dunn’s test were used to compare median costs between the groups and Spearman's correlation test was used for correlation studies. Results: Uncomplicated patients had significantly lower costs (p <0.0001) compared to other groups. Patients with IHD had highest Medical expenses (p < 0.0001), followed by DN and DF (p < 0.0001 ). Annual medical costs incurred were 1.8, 2.76, 2.77, 1.76, and 4.34 times higher in retinopathy, nephropathy, diabetic foot, neuropathy and IHD patients as compared to the cost incurred in managing uncomplicated diabetics. Other costs also showed a similar pattern of rising. A positive correlation was observed between the costs incurred and duration of diabetes, a negative correlation between the glycemic status and cost incurred. The cost incurred in the management of DM in 2017 was found to be elevated 1.4 - 2.7 times when compared to that in 2008. Conclusion: It is evident from the study that the economic burden due to diabetes mellitus is substantial. It poses a significant financial burden on the healthcare system, individual and society as a whole. There is a need for the strategies to achieve optimal glycemic control and operationalize regular and early screening methods for complications so as to reduce the burden of the disease.

Keywords: COI, diabetes mellitus, a bottom up approach, economics

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Authors: Chu-Tzu Chen, Angela S. Huang, Yu-Min Chou, Chin-Hui Yang

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Dengue is a major public health concern throughout tropical and sub-tropical regions. Taiwan is located in the Pacific Ocean and overlying the tropical and subtropical zones. The island remains humid throughout the year and receives abundant rainfall, and the temperature is very hot in summer at southern Taiwan. It is ideal for the growth of dengue vectors and would be increasing the risk on dengue outbreaks. During the first half of the 20th century, there were three island-wide dengue outbreaks (1915, 1931, and 1942). After almost forty years of dormancy, a DEN-2 outbreak occurred in Liuchiu Township, Pingtung County in 1981. Thereafter, more dengue outbreaks occurred with different scales in southern Taiwan. However, there were more than ten thousands of dengue cases in 2014 and in 2015. It did not only affect human health, but also caused widespread social disruption and economic losses. The study would like to reveal the epidemiology of dengue on Taiwan, especially the severe outbreak in 2015, and try to find the effective interventions in dengue control including dengue vaccine development for the elderly. Methods: The study applied the Notifiable Diseases Surveillance System database of the Taiwan Centers for Disease Control as data source. All cases were reported with the uniform case definition and confirmed by NS1 rapid diagnosis/laboratory diagnosis. Results: In 2014, Taiwan experienced a serious DEN-1 outbreak with 15,492 locally-acquired cases, including 136 cases of dengue hemorrhagic fever (DHF) which caused 21 deaths. However, a more serious DEN-2 outbreak occurred with 43,419 locally-acquired cases in 2015. The epidemic occurred mainly at Tainan City (22,760 cases) and Kaohsiung City (19,723 cases) in southern Taiwan. The age distribution for the cases were mainly adults. There were 228 deaths due to dengue infection, and the case fatality rate was 5.25 ‰. The average age of them was 73.66 years (range 29-96) and 86.84% of them were older than 60 years. Most of them were comorbidities. To review the clinical manifestations of the 228 death cases, 38.16% (N=87) of them were reported with warning signs, while 51.75% (N=118) were reported without warning signs. Among the 87 death cases reported to dengue with warning signs, 89.53% were diagnosed sever dengue and 84% needed the intensive care. Conclusion: The year 2015 was characterized by large dengue outbreaks worldwide. The risk of serious dengue outbreak may increase significantly in the future, and the elderly is the vulnerable group in Taiwan. However, a dengue vaccine has been licensed for use in people 9-45 years of age living in endemic settings at the end of 2015. In addition to carry out the research to find out new interventions in dengue control, developing the dengue vaccine for the elderly is very important to prevent severe dengue and deaths.

Keywords: case fatality rate, dengue, dengue vaccine, the elderly

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Authors: Olga Gomez De Miranda, Jose R. Ochoa-Gomez, Stefaan De Wildeman, Luciano Monsegue, Soraya Prieto, Leire Lorenzo, Cristina Dineiro

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The chemical industry is facing a new revolution. As long as processes based on the exploitation of fossil resources emerged with force in the XIX century, Society currently demands a new radical change that will lead to the complete and irreversible implementation of a circular sustainable economic model. The implementation of biorefineries will be essential for this. There, renewable raw materials as sugars and other biomass resources are exploited for the development of new materials that will partially replace their petroleum-derived homologs in a safer, and environmentally more benign approach. Isosorbide, (1,4:3,6-dianhydro-d-glucidol) is a primary bio-based derivative obtained from the plant (poly) saccharides and a very interesting example of a useful chemical produced in biorefineries. It can, in turn, be converted to other secondary monomers as isosorbide bis-methyl carbonate (IBMC), whose main field of application can be as a key biodegradable intermediary substitute of bisphenol-A in the manufacture of polycarbonates, or as an alternative to the toxic isocyanates in the synthesis of new polyurethanes (non-isocyanate polyurethanes) both with a huge application market. New products will present advantageous mechanical or optical properties, as well as improved behavior in non-toxicity and biodegradability aspects in comparison to their petro-derived alternatives. A robust production process of IBMC, a biomass-derived chemical, is here presented. It can be used with different raw material qualities using dimethyl carbonate (DMC) as both co-reactant and solvent. It consists of the transesterification of isosorbide with DMC under soft operational conditions, using different basic catalysts, always active with the isosorbide characteristics and purity. Appropriate isolation processes have been also developed to obtain crude IBMC yields higher than 90%, with oligomers production lower than 10%, independently of the quality of the isosorbide considered. All of them are suitable to be used in polycondensation reactions for polymers obtaining. If higher qualities of IBMC are needed, a purification treatment based on nanofiltration membranes has been also developed. The IBMC reaction-isolation conditions established in the laboratory have been successfully modeled using appropriate software programs and moved to a pilot-scale (production of 100 kg of IBMC). It has been demonstrated that a highly efficient IBMC production process able to be up-scaled under suitable market conditions has been obtained. Operational conditions involved the production of IBMC involve soft temperature and energy needs, no additional solvents, and high operational efficiency. All of them are according to green manufacturing rules.

Keywords: biomass, catalyst, isosorbide bis-methyl carbonate, polycarbonate, polyurethane, transesterification

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Authors: Takaaki Nemoto

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Case presentation: A 69-year-old Japanese man presented with a low-grade fever and fatigue that had persisted for one month. The patient had an aortic dissection on the aortic arch 13 years prior, an abdominal aortic aneurysm seven years prior, and an aortic dissection on the distal aortic arch one year prior, which were all treated with artificial blood-vessel replacement surgery. Laboratory tests revealed an inflammatory response (CRP 7.61 mg/dl), high serum creatinine (Cr 1.4 mg/dL), and elevated transaminase (AST 47 IU/L, ALT 45 IU/L). The patient was admitted to our hospital on suspicion of prosthetic vascular graft infection. Following further workups on the inflammatory response, an enhanced chest computed tomography (CT) and a non-enhanced chest DWI (MRI) were performed. The patient was diagnosed with a pulmonary fistula and a prosthetic vascular graft infection on the distal aortic arch. After admission, the patient was administered Ceftriaxion and Vancomycine for 10 days, but his fever and inflammatory response did not improve. On day 13 of hospitalization, a lung fistula repair surgery and an omental filling operation were performed, and Meropenem and Vancomycine were administered. The fever and inflammatory response continued, and therefore we took repeated blood cultures. M. fortuitum was detected in a blood culture on day 16 of hospitalization. As a result, we changed the treatment regimen to Amikacin (400 mg/day), Meropenem (2 g/day), and Cefmetazole (4 g/day), and the fever and inflammatory response began to decrease gradually. We performed a test of sensitivity for Mycobacterium fortuitum, and found that the MIC was low for fluoroquinolone antibacterial agent. The clinical course was good, and the patient was discharged after a total of 8 weeks of intravenous drug administration. At discharge, we changed the treatment regimen to Levofloxacin (500 mg/day) and Clarithromycin (800 mg/day), and prescribed these two drugs as a long life suppressive therapy. Discussion: There are few cases of prosthetic vascular graft infection caused by mycobacteria, and a standard therapy remains to be established. For prosthetic vascular graft infections, it is ideal to provide surgical and medical treatment in parallel, but in this case, surgical treatment was difficult and, therefore, a conservative treatment was chosen. We attempted to increase the treatment success rate of this refractory disease by conducting a susceptibility test for mycobacteria and treating with different combinations of antimicrobial agents, which was ultimately effective. With our treatment approach, a good clinical course was obtained and continues at the present stage. Conclusion: Although prosthetic vascular graft infection resulting from mycobacteria is a refractory infectious disease, it may be curative to administer appropriate antibiotics based on the susceptibility test in addition to surgical treatment.

Keywords: prosthetic vascular graft infection, lung fistula, Mycobacterium fortuitum, conservative treatment

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Authors: Ciprian Briciu-Burghina, Brendan Heery, Dermot Brabazon, Fiona Regan

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Good quality bathing water is a highly desirable natural resource which can provide major economic, social, and environmental benefits. Both in Ireland and Europe, such water bodies are managed under the European Directive for the management of bathing water quality (BWD). The BWD aims mainly: (i) to improve health protection for bathers by introducing stricter standards for faecal pollution assessment (E. coli, enterococci), (ii) to establish a more pro-active approach to the assessment of possible pollution risks and the management of bathing waters, and (iii) to increase public involvement and dissemination of information to the general public. Standard methods for E. coli and enterococci quantification rely on cultivation of the target organism which requires long incubation periods (from 18h to a few days). This is not ideal when immediate action is required for risk mitigation. Municipalities that oversee the bathing water quality and deploy appropriate signage have to wait for laboratory results. During this time, bathers can be exposed to pollution events and health risks. Although forecasting tools exist, they are site specific and as consequence extensive historical data is required to be effective. Another approach for early detection of faecal pollution is the use of marker enzymes. β-glucuronidase (GUS) is a widely accepted biomarker for E. coli detection in microbiological water quality control. GUS assay is particularly attractive as they are rapid, less than 4 h, easy to perform and they do not require specialised training. A method for on-site detection of GUS from environmental samples in less than 75 min was previously demonstrated. In this study, the capability of ColiSense as an early warning system for faecal pollution in freshwater is assessed. The system successfully detected GUS activity in all of the 45 freshwater samples tested. GUS activity was found to correlate linearly with E. coli (r2=0.53, N=45, p < 0.001) and enterococci (r2=0.66, N=45, p < 0.001) Although GUS is a marker for E. coli, a better correlation was obtained for enterococci. For this study water samples were collected from 5 rivers in the Dublin area over 1 month. This suggests a high diversity of pollution sources (agricultural, industrial, etc) as well as point and diffuse pollution sources were captured in the sample size. Such variety in the source of E. coli can account for different GUS activities/culturable cell and different ratios of viable but not culturable to viable culturable bacteria. A previously developed protocol for the recovery and detection of E. coli was coupled with a miniaturised fluorometer (ColiSense) and the system was assessed for the rapid detection FIB in freshwater samples. Further work will be carried out to evaluate the system’s performance on seawater samples.

Keywords: faecal pollution, β-glucuronidase (GUS), bathing water, E. coli

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Authors: Ivana Loncarevic, Biljana Pajin, Jovana Petrovic, Danica Zaric, Vesna Tumbas Saponjac, Aleksandar Fistes

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Chocolate is a food that activates pleasure centers in the human brain. In comparison to black and milk chocolate, white chocolate does not contain fat-free cocoa solids and thus lacks bioactive components. The aim of this study was to examine the sensory characteristics of enriched white chocolate with the addition of 10% of raspberry juice encapsulated in maltodextrins (denoted as encapsulate). Chocolate is primarily intended for enjoyment, and therefore, the sensory expectation is a critical factor for consumers when selecting a new type of chocolate. Consumer acceptance of chocolate depends primarily on the appearance and taste, but also very much on the mouthfeel, which mainly depends on the particle size of chocolate. Chocolate samples were evaluated by a panel of 8 trained panelists, food technologists, trained according to ISO 8586 (2012). Panelists developed the list of attributes to be used in this study: intensity of red color (light to dark); glow on the surface (mat to shiny); texture on snap (appearance of cavities or holes on the snap surface that are seen - even to gritty); hardness (hardness felt during the first bite of chocolate sample in half by incisors - soft to hard); melting (the time needed to convert solid chocolate into a liquid state – slowly to quickly); smoothness (perception of evenness of chocolate during melting - very even to very granular); fruitiness (impression of fruity taste - light fruity notes to distinct fruity notes); sweetness (organoleptic characteristic of pure substance or mixture giving sweet taste - lightly sweet to very sweet). The chocolate evaluation was carried out 24 h after sample preparation in the sensory laboratory, in partitioned booths, which were illuminated with fluorescent lights (ISO 8589, 2007). Samples were served in white plastic plates labeled with three-digit codes from a random number table. Panelist scored the perceived intensity of each attribute using a 7-point scale (1 = the least intensity and 7 = the most intensity) (ISO 4121, 2002). The addition of 10% of encapsulate had a big influence on chocolate color, where enriched chocolate got a nice reddish color. At the same time, the enriched chocolate sample had less intensity of gloss on the surface. The panelists noticed that addition of encapsulate reduced the time needed to convert solid chocolate into a liquid state, increasing its hardness. The addition of encapsulate had a significant impact on chocolate flavor. It reduced the sweetness of white chocolate and contributed to the fruity raspberry flavor.

Keywords: white chocolate, encapsulated raspberry juice, color, sensory characteristics

Procedia PDF Downloads 160

Authors: Natsuki Kishizawa, Keiko Nakano, Hussam Organji, Amer Shaiban, Mohammad Albeirutty

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One of the most important tasks in operating desalination plants using a reverse osmosis (RO) method is preventing RO membrane fouling caused by foulants found in seawater. Optimal design of the pre-treatment process of RO process for plants enables the reduction of foulants. Therefore, a quantitative evaluation of the fouling risk in pre-treated water, which is fed to RO, is required for optimal design. Some measurement methods for water quality such as silt density index (SDI) and total organic carbon (TOC) have been conservatively applied for evaluations. However, these methods have not been effective in some situations for evaluating the fouling risk of RO feed water. Furthermore, stable management of plants will be possible by alerts and appropriate control of the pre-treatment process by using the method if it can be applied to the inline monitoring system for the fouling risk of RO feed water. The purpose of this study is to develop a method to evaluate the fouling risk of RO feed water. We applied a quartz crystal microbalance (QCM) to measure the amount of foulants found in seawater using a sensor whose surface is coated with polyamide thin film, which is the main material of a RO membrane. The increase of the weight of the sensor after a certain length of time in which the sample water passes indicates the fouling risk of the sample directly. We classified the values as “FP: Fouling Potential”. The characteristics of the method are to measure the very small amount of substances in seawater in a short time: < 2h, and from a small volume of the sample water: < 50mL. Using some RO cell filtration units, a higher correlation between the pressure increase given by RO fouling and the FP from the method than SDI and TOC was confirmed in the laboratory-scale test. Then, to establish the correlation in the actual bench-scale RO membrane module, and to confirm the feasibility of the monitoring system as a control tool for the pre-treatment process, we have started a long-term test at an experimental desalination site by the Red Sea in Jeddah, Kingdom of Saudi Arabia. Implementing inline equipment for the method made it possible to measure FP intermittently (4 times per day) and automatically. Moreover, for two 3-month long operations, the RO operation pressure among feed water samples of different qualities was compared. The pressure increase through a RO membrane module was observed at a high FP RO unit in which feed water was treated by a cartridge filter only. On the other hand, the pressure increase was not observed at a low FP RO unit in which feed water was treated by an ultra-filter during the operation. Therefore, the correlation in an actual scale RO membrane was established in two runs of two types of feed water. The result suggested that the FP method enables the evaluation of the fouling risk of RO feed water.

Keywords: fouling, monitoring, QCM, water quality

Procedia PDF Downloads 212

Authors: Kristina Amaleviciute, Ieva Jokubauskaite, Alvyra Slepetiene, Jonas Volungevicius, Inga Liaudanskiene

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The main task of this research was to investigate the chemical composition of the differently used soil in profiles. To identify the differences in the soil were investigated organic carbon (SOC) and its fractional composition: dissolved organic carbon (DOC), mobile humic acids (MHA) and C to N ratio of natural and anthropogenically affected Luvisols. Research object: natural and anthropogenically affected Luvisol, Akademija, Kedainiai, distr. Lithuania. Chemical analyses were carried out at the Chemical Research Laboratory of Institute of Agriculture, LAMMC. Soil samples for chemical analyses were taken from the genetics soil horizons. SOC was determined by the Tyurin method modified by Nikitin, measuring with spectrometer Cary 50 (VARIAN) in 590 nm wavelength using glucose standards. For mobile humic acids (MHA) determination the extraction procedure was carried out using 0.1 M NaOH solution. Dissolved organic carbon (DOC) was analyzed using an ion chromatograph SKALAR. pH was measured in 1M H2O. N total was determined by Kjeldahl method. Results: Based on the obtained results, it can be stated that transformation of chemical composition is going through the genetic soil horizons. Morphology of the upper layers of soil profile which is formed under natural conditions was changed by anthropomorphic (agrogenic, urbogenic, technogenic and others) structure. Anthropogenic activities, mechanical and biochemical disturbances destroy the natural characteristics of soil formation and complicates the interpretation of soil development. Due to the intensive cultivation, the pH values of the curve equals (disappears acidification characteristic for E horizon) with natural Luvisol. Luvisols affected by agricultural activities was characterized by a decrease in the absolute amount of humic substances in separate horizons. But there was observed more sustainable, higher carbon sequestration and thicker storage of humic horizon compared with forest Luvisol. However, the average content of humic substances in the soil profile was lower. Soil organic carbon content in anthropogenic Luvisols was lower compared with the natural forest soil, but there was more evenly spread over in the wider thickness of accumulative horizon. These data suggest that the organization of geo-ecological declines and agroecological increases in Luvisols. Acknowledgement: This work was supported by the National Science Program ‘The effect of long-term, different-intensity management of resources on the soils of different genesis and on other components of the agro-ecosystems’ [grant number SIT-9/2015] funded by the Research Council of Lithuania.

Keywords: agrogenization, dissolved organic carbon, luvisol, mobile humic acids, soil organic carbon

Procedia PDF Downloads 237

Authors: Alexandre Faure, Yoko Mizokami, éRic Dinet

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These recent years, the potential of light in assisting visually impaired people in their indoor mobility has been demonstrated by different studies. Implementing smart lighting systems for selective visual enhancement, especially designed for low-vision people, is an approach that breaks with the existing visual aids. The appearance of the surface of an object is significantly influenced by the lighting conditions and the constituent materials of the objects. Appearance of objects may appear to be different from expectation. Therefore, lighting conditions lead to an important part of accurate material recognition. The main objective of this work was to investigate the effect of the spatial distribution of light on object identification in the context of low vision. The purpose was to determine whether and what specific lighting approaches should be preferred for visually impaired people. A psychophysical experiment was designed to study the ability of individuals to identify the smallest cube of a pair under different lighting diffusion conditions. Participants were divided into two distinct groups: a reference group of observers with normal or corrected-to-normal visual acuity and a test group, in which observers were required to wear visual impairment simulation glasses. All participants were presented with pairs of cubes in a "miniature room" and were instructed to estimate the relative size of the two cubes. The miniature room replicates real-life settings, adorned with decorations and separated from external light sources by black curtains. The correlated color temperature was set to 6000 K, and the horizontal illuminance at the object level at approximately 240 lux. The objects presented for comparison consisted of 11 white cubes and 11 black cubes of different sizes manufactured with a 3D printer. Participants were seated 60 cm away from the objects. Two different levels of light diffuseness were implemented. After receiving instructions, participants were asked to judge whether the two presented cubes were the same size or if one was smaller. They provided one of five possible answers: "Left one is smaller," "Left one is smaller but unsure," "Same size," "Right one is smaller," or "Right one is smaller but unsure.". The method of constant stimuli was used, presenting stimulus pairs in a random order to prevent learning and expectation biases. Each pair consisted of a comparison stimulus and a reference cube. A psychometric function was constructed to link stimulus value with the frequency of correct detection, aiming to determine the 50% correct detection threshold. Collected data were analyzed through graphs illustrating participants' responses to stimuli, with accuracy increasing as the size difference between cubes grew. Statistical analyses, including 2-way ANOVA tests, showed that light diffuseness had no significant impact on the difference threshold, whereas object color had a significant influence in low vision scenarios. The first results and trends derived from this pilot experiment clearly and strongly suggest that future investigations could explore extreme diffusion conditions to comprehensively assess the impact of diffusion on object identification. For example, the first findings related to light diffuseness may be attributed to the range of manipulation, emphasizing the need to explore how other lighting-related factors interact with diffuseness.

Keywords: Lighting, Low Vision, Visual Aid, Object Identification, Psychophysical Experiment

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Authors: Tomas Makaras, Gintaras Svecevičius

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Landfill waste is a common problem as it has an economic and environmental impact even if it is closed. Landfill waste contains a high density of various persistent compounds such as heavy metals, organic and inorganic materials. As persistent compounds are slowly-degradable or even non-degradable in the environment, they often produce sublethal or even lethal effects on aquatic organisms. The aims of the present study were to estimate sublethal effects of the Kairiai landfill (WGS: 55°55‘46.74“, 23°23‘28.4“) leachate on the locomotor activity of rainbow trout Oncorhynchus mykiss juveniles using the original system package developed in our laboratory for automated monitoring, recording and analysis of aquatic organisms’ activity, and to determine patterns of fish behavioral response to sublethal effects of leachate. Four different concentrations of leachate were chosen: 0.125; 0.25; 0.5 and 1.0 mL/L (0.0025; 0.005; 0.01 and 0.002 as part of 96-hour LC50, respectively). Locomotor activity was measured after 5, 10 and 30 minutes of exposure during 1-minute test-periods of each fish (7 fish per treatment). The threshold-effect-concentration amounted to 0.18 mL/L (0.0036 parts of 96-hour LC50). This concentration was found to be even 2.8-fold lower than the concentration generally assumed to be “safe” for fish. At higher concentrations, the landfill leachate solution elicited behavioral response of test fish to sublethal levels of pollutants. The ability of the rainbow trout to detect and avoid contaminants occurred after 5 minutes of exposure. The intensity of locomotor activity reached a peak within 10 minutes, evidently decreasing after 30 minutes. This could be explained by the physiological and biochemical adaptation of fish to altered environmental conditions. It has been established that the locomotor activity of juvenile trout depends on leachate concentration and exposure duration. Modeling of these parameters showed that the activity of juveniles increased at higher leachate concentrations, but slightly decreased with the increasing exposure duration. Experiment results confirm that the behavior of rainbow trout juveniles is a sensitive and rapid biomarker that can be used in combination with the system for fish behavior monitoring, registration and analysis to determine sublethal concentrations of pollutants in ambient water. Further research should be focused on software improvement aimed to include more parameters of aquatic organisms’ behavior and to investigate the most rapid and appropriate behavioral responses in different species. In practice, this study could be the basis for the development and creation of biological early-warning systems (BEWS).

Keywords: fish behavior biomarker, landfill leachate, locomotor activity, rainbow trout juveniles, sublethal effects

Procedia PDF Downloads 273

Authors: Mustafa M. Donma, Orkide Donma

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Obesity is known to be associated with many clinically important diseases including metabolic syndrome (MetS). Vitamin B₁₂ plays essential roles in fat and protein metabolisms and its cooperation with vitamin B₉ is well-known. The aim of this study is to investigate the possible contributions as well as associations of these micronutrients upon obesity and MetS during childhood. A total of 128 children admitted to Namik Kemal University, Medical Faculty, Department of Pediatrics Outpatient Clinics were included into the scope of this study. The mean age±SEM of 92 morbid obese (MO) children and 36 with MetS were 118.3±3.8 months and 129.5±6.4 months, respectively (p > 0.05). The study was approved by Namık Kemal University, Medical Faculty Ethics Committee. Written informed consent forms were obtained from the parents. Demographic features and anthropometric measurements were recorded. WHO BMI-for age percentiles were used. The values above 99 percentile were defined as MO. Components of MetS [waist circumference (WC), fasting blood glucose (FBG), triacylglycerol (TRG), high density lipoprotein cholesterol (HDL-Chol), systolic pressure (SP), diastolic pressure (DP)] were determined. Routine laboratory tests were performed. Serum vitamin B₁₂ concentrations were measured using electrochemiluminescence immunoassay. Vitamin B₉ was analyzed by an immunoassay analyzer. Values for vitamin B₁₂ < 148 pmol/L, 148-221 pmol/L, > 221 pmol/L were accepted as low, borderline and normal, respectively. Vitamin B₉ levels ≤ 4 mcg/L defined deficiency state. Statistical evaluations were performed by SPSSx Version 16.0. p≤0.05 was accepted as statistical significance level. Statistically higher body mass index (BMI), WC, hip circumference (C) and neck C were calculated in MetS group compared to children with MO. No difference was noted for head C. All MetS components differed between the groups (SP, DP p < 0.001; WC, FBG, TRG p < 0.01; HDL-Chol p < 0.05). Significantly decreased vitamin B₉ and vitamin B₁₂ levels were detected (p < 0.05) in children with MetS. In both groups percentage of folate deficiency was 5.5%. No cases were below < 148 pmol/L. However, in MO group 14.3% and in MetS group 22.2% of the cases were of borderline status. In MO group B₁₂ levels were negatively correlated with BMI, WC, hip C and head C, but not with neck C. WC, hip C, head C and neck C were all negatively correlated with HDL-Chol. None of these correlations were observed in the group of children with MetS. Strong positive correlation between FBG and insulin as well as strong negative correlation between TRG and HDL-Chol detected in MO children were lost in MetS group. Deficiency state end-products of both B₉ and B₁₂ may interfere with the expected profiles of MetS components. In this study, the alterations in MetS components affected vitamin B₁₂ metabolism and also its associations with anthropometric body measurements. Further increases in vitamin B₁₂ and vitamin B₉ deficiency in MetS associated with the increased vitamin B₁₂ as well as vitamin B₉ deficiency metabolites may add to MetS parameters.

Keywords: children, cobalamin, folate, metabolic syndrome, obesity

Procedia PDF Downloads 194

Authors: Seyed Hamed Mousavi, Juha M. Hijmans, Reza Rajabi, Ron Diercks, Johannes Zwerver, Henk van der Worp

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Introduction: Running is one of the most popular sports activity among people. Improper sagittal plane ankle, knee and hip kinematics are considered to be associated with the increase of injury risk in runners. Motion assessing smart-phone applications are increasingly used to measure kinematics both in the field and laboratory setting, as they are cheaper, more portable, accessible, and easier to use relative to 3D motion analysis system. The aims of this study are 1) to compare the results of 3D gait analysis system and CE; 2) to evaluate the test-retest and intra-rater reliability of coach’s eye (CE) app for the sagittal plane hip, knee, and ankle angles in the touchdown and toe-off while running. Method: Twenty subjects participated in this study. Sixteen reflective markers and cluster markers were attached to the subject’s body. Subjects were asked to run at a self-selected speed on a treadmill. Twenty-five seconds of running were collected for analyzing kinematics of interest. To measure sagittal plane hip, knee and ankle joint angles at touchdown (TD) and toe off (TO), the mean of first ten acceptable consecutive strides was calculated for each angle. A smartphone (Samsung Note5, android) was placed on the right side of the subject so that whole body was simultaneously filmed with 3D gait system during running. All subjects repeated the task with the same running speed after a short interval of 5 minutes in between. The CE app, installed on the smartphone, was used to measure the sagittal plane hip, knee and ankle joint angles at touchdown and toe off the stance phase. Results: Intraclass correlation coefficient (ICC) was used to assess test-retest and intra-rater reliability. To analyze the agreement between 3D and 2D outcomes, the Bland and Altman plot was used. The values of ICC were for Ankle at TD (TRR=0.8,IRR=0.94), ankle at TO (TRR=0.9,IRR=0.97), knee at TD (TRR=0.78,IRR=0.98), knee at TO (TRR=0.9,IRR=0.96), hip at TD (TRR=0.75,IRR=0.97), hip at TO (TRR=0.87,IRR=0.98). The Bland and Altman plots displaying a mean difference (MD) and ±2 standard deviation of MD (2SDMD) of 3D and 2D outcomes were for Ankle at TD (MD=3.71,+2SDMD=8.19, -2SDMD=-0.77), ankle at TO (MD=-1.27, +2SDMD=6.22, -2SDMD=-8.76), knee at TD (MD=1.48, +2SDMD=8.21, -2SDMD=-5.25), knee at TO (MD=-6.63, +2SDMD=3.94, -2SDMD=-17.19), hip at TD (MD=1.51, +2SDMD=9.05, -2SDMD=-6.03), hip at TO (MD=-0.18, +2SDMD=12.22, -2SDMD=-12.59). Discussion: The ability that the measurements are accurately reproduced is valuable in the performance and clinical assessment of outcomes of joint angles. The results of this study showed that the intra-rater and test-retest reliability of CE app for all kinematics measured are excellent (ICC ≥ 0.75). The Bland and Altman plots display that there are high differences of values for ankle at TD and knee at TO. Measuring ankle at TD by 2D gait analysis depends on the plane of movement. Since ankle at TD mostly occurs in the none-sagittal plane, the measurements can be different as foot progression angle at TD increases during running. The difference in values of the knee at TD can depend on how 3D and the rater detect the TO during the stance phase of running.

Keywords: reliability, running, sagittal plane, two dimensional

Procedia PDF Downloads 202

Authors: Izabela Poprawa, Alina Chachulska-Zymelka, Lukasz Chajec, Grazyna Wilczek, Piotr Wilczek, Sebastian Student, Magdalena Rost-Roszkowska

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Heavy metals polluting the environment, especially soil, have a harmful effect on organisms, because they can damage the organ structure, disturb their function and cause developmental disorders. They can affect not only the somatic tissues but also the germinal tissues. In the natural environment, plants and animals are exposed to short- and long-term exposure to these stressors, which have a major influence on the functioning of these organisms. Numerous animals have been treated as the bioindicators of the environment. Therefore, studies on any alterations caused by, e.g., heavy metals are in the center of interests of not only environmental but also medical and biological science. Myriapods are invertebrates which are bioindicators of the environment. One of the species which lives in the upper layers of soil, particularly under stones and rocks is Lithobius forficatus (Chilopoda), commonly known as the brown centipede or stone centipede. It is a European species of the family Lithobiidae. This centipede living in the soil is exposed to, e.g., heavy metals such as cadmium, lead, arsenic. The main goal of our project was to analyze the impact of long-term exposure to cadmium on the structure of ovary with the emphasis on the course of oogenesis. As the material for analysis of cadmium exposure to ovaries, we chose the centipede species, L. forficatus. Animals were divided into two experimental groups: C – the control group, the animals cultured in laboratory conditions in a horticultural soil; Cd2 – the animals cultured in a horticultural soil supplemented with 80 mg/kg (dry weight) of CdCl2 for 45 days – long-term exposure. Animals were fed with Acheta and Chironomus larvae maintained in tap water. The analyzes were carried out using transmission electron microscopy (TEM), flow cytometry and laser scanning (confocal) microscopy. Here we present the results of long-term exposure to cadmium concentration in soil on the organ responsible for female germ cell formation. Analysis with the use of the transmission electron microscope showed changes in the ultrastructure of both somatic and germ cells in the ovary. Moreover, quantitative analysis revealed the decrease in the percentage of cells viability, the increase in the percentage of cells with depolarized mitochondria and increasing the number of early apoptotic cells. All these changes were statistically significant compared to the control. Additionally, an increase in the ADP/ATP index was recorded. However, changes were not statistically significant to the control. Acknowledgment: The study has been financed by the National Science Centre, Poland, grant no 2017/25/B/NZ4/00420.

Keywords: cadmium, centipede, ovary, ultrastructure

Procedia PDF Downloads 118

Authors: Rensson Homero Celiz Ygnacio, Marco Aurélio Schiavo Novaes, Lucy Vanessa Sulca Ñaupas, Ana Paula Ribeiro Rodrigues

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The cryopreservation of testicular tissue emerges as an alternative for the preservation of the reproductive potential of individuals who still cannot produce sperm; however, they will undergo treatments that may affect their fertility (e.g., chemotherapy). Therefore, the present work aims to compare two cryopreservation methods (slow freezing and vitrification) in testicular tissue of prepubertal lambs. For that, to obtain the testicular tissue, the animals were castrated and the testicles were collected immediately in a physiological solution supplemented with antibiotics. In the laboratory, the testis was split into small pieces. The total size of the testicular fragments was 3×3x1 mm³ and was placed in a dish contained in Minimum Essential Medium (MEM-HEPES). The fragments were distributed randomly into non-cryopreserved (fresh control), slow freezing (SF), and vitrified. To SF procedures, two fragments from a given male were then placed in a 2,0 mL cryogenic vial containing 1,0 mL MEM-HEPES supplemented with 20% fetal bovine serum (FBS) and 20% dimethylsulfoxide (DMSO). Tubes were placed into a Mr. Frosty™ Freezing container with isopropyl alcohol and transferred to a -80°C freezer for overnight storage. On the next day, each tube was plunged into liquid nitrogen (NL). For vitrification, the ovarian tissue cryosystem (OTC) device was used. Testicular fragments were placed in the OTC device and exposed to the first vitrification solution composed of MEM-HEPES supplemented with 10 mg/mL Bovine Serum Albumin (BSA), 0.25 M sucrose, 10% Ethylene glycol (EG), 10% DMSO and 150 μM alpha-lipoic acid for four min. The VS1 was discarded and then the fragments were submerged into a second vitrification solution (VS2) containing the same composition of VS1 but 20% EG and 20% DMSO. VS2 was then discarded and each OTC device containing up to four testicular fragments was closed and immersed in NL. After the storage period, the fragments were removed from the NL, kept at room temperature for one min and then immersed at 37 °C in a water bath for 30 s. Samples were warmed by sequentially immersing in solutions of MEM-HEPES supplemented with 3 mg/mL BSA and decreasing concentrations of sucrose. Hematoxylin-eosin staining to analyze the tissue architecture was used. The score scale used was from 0 to 3, classified with a score 0 representing normal morphologically, and 3 were considered a lot of alteration. The histomorphological evaluation of the testicular tissue shows that when evaluating the nuclear alteration (distinction of nucleoli and condensation of nuclei), there are no differences when using slow freezing with respect to the control. However, vitrification presents greater damage (p <0.05). On the other hand, when evaluating the epithelial alteration, we observed that the freezing showed scores statistically equal to the control in variables such as retraction of the basement membrane, formation of gaps and organization of the peritubular cells. The results of the study demonstrated that cryopreservation using the slow freezing method is an excellent tool for the preservation of pubertal testicular tissue.

Keywords: cryopreservation, slow freezing, vitrification, testicular tissue, lambs

Procedia PDF Downloads 175

Authors: L. C. Sánchez, L. C. Corrales, A. G. Lancheros, E. Castañeda, Y. Ariza, L. S. Fuentes, L. Sierra, J. L. Cuervo

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The importance of environment friendly alternatives in agricultural processes is the reason why the research group Ceparium, the Colegio Mayor de Cundinamarca University, Colombia, investigated the genus Bacillus and its applicability for improving crops of economic importance in Colombia. In this investigation, we presented a study in which the genus Bacillus plays a leading role as beneficial microorganism. The objective was to identify the biochemical potential of three indigenous species of Bacillus, which were able to carry out actions for biological control against pathogens and pests or promoted growth to improve productivity of crops in Colombia. The procedures were performed in three phases: first, the production of biomass of an indigenous strain and a reference strain starting from culture media for production of spores and toxins were made. Spore count was done in a Neubauer chamber, concentrations of spores of Bacillus sphaericus were prepared and a bioassay was done at the Laboratory of Entomology at the University Jorge Tadeo Lozano of Plutella xylostella larvae, insect pest of crucifers in several Colombian regions. The second phase included the extraction in the liquid state fermentation, a secondary metabolite that has antibiosis action against fungi, call iturin B, and was obtained from strains of Bacillus subtilis. The molecule was identified using High Resolution Chromatography (HPLC) and its biocontrol effect on Fusarium sp fungus causes vascular wilt in economically important plant varieties, was confirmed using testing of antagonism in Petri dish. In the third phase, an initial procedure in that let recover and identify microorganisms of the genus Bacillus from the rhizosphere in two aromatic herbs, Rosmarinus officinalis and Thymus vulgaris L. was used. Subsequently, testing of antagonism against Fusarium sp were made and an assay was done under greenhouse conditions to observe biocontrol and growth promoting action by comparing growth in length and dry weight. In the first experiment, native Bacillus sphaericus was lethal to 92% Plutella xylostella larvae in 10 DDA. In the second experiment, iturin B was identified and biological control of Fusarium sp was demonstrated. In the third study, all strains demonstrated biological control and the B14 strain identified as Bacillus megaterium increased root length and productivity of the two plants in terms of weight. It was concluded that the native microorganisms of the genus Bacillus has a great biochemical potential that provides a beneficial interactions with plants, improve their growth and development and therefore a greater impact on production.

Keywords: genus bacillus, biological control, PGPRs, biochemical potential

Procedia PDF Downloads 436

Authors: Nor Azian Binti Aziz, Muhammad Afiq Bin Tambichik, Zamri Bin Hashim

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The rapid development of the construction industry has contributed to increased construction waste, with concrete waste being among the most abundant. This waste is generated from ready-mix batching plants after the concrete cube testing process is completed and disposed of in landfills, leading to increased solid waste management costs. This study aims to evaluate the engineering characteristics of foamed concrete with waste mixtures construction and agricultural waste to determine the usability of recycled materials in the construction of non-load-bearing walls. This study involves the collection of construction wastes, such as recycled aggregates (RCA) obtained from the remains of finished concrete cubes, which are then tested in the laboratory. Additionally, agricultural waste, such as rice husk ash, is mixed into foamed concrete interlock blocks to enhance their strength. The optimal density of foamed concrete for this study was determined by mixing mortar and foam-backed agents to achieve the minimum targeted compressive strength required for non-load-bearing walls. The tests conducted in this study involved two phases. In Phase 1, elemental analysis using an X-ray fluorescence spectrometer (XRF) was conducted on the materials used in the production of interlock blocks such as sand, recycled aggregate/recycled concrete aggregate (RCA), and husk ash paddy/rice husk ash (RHA), Phase 2 involved physical and thermal tests, such as compressive strength test, heat conductivity test, and fire resistance test, on foamed concrete mixtures. The results showed that foamed concrete can produce lightweight interlock blocks. X-ray fluorescence spectrometry plays a crucial role in the characterization, quality control, and optimization of foamed concrete mixes containing construction and agriculture waste. The unique composition mixer of foamed concrete and the resulting chemical and physical properties, as well as the nature of replacement (either as cement or fine aggregate replacement), the waste contributes differently to the performance of foamed concrete. Interlocking blocks made from foamed concrete can be advantageous due to their reduced weight, which makes them easier to handle and transport compared to traditional concrete blocks. Additionally, foamed concrete typically offers good thermal and acoustic insulation properties, making it suitable for a variety of building projects. Using foamed concrete to produce lightweight interlock blocks could contribute to more efficient and sustainable construction practices. Additionally, RCA derived from concrete cube waste can serve as a substitute for sand in producing lightweight interlock blocks.

Keywords: construction waste, recycled aggregates (RCA), sustainable concrete, structure material

Procedia PDF Downloads 55

Authors: E. Tuğba Topçu, Ebru E. Kazan, Erhan Büken

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Emergency nurses are the first health care professional to generally observe the patients, communicate patients’ family or relatives, touch the properties of patients and contact to laboratory sample of patients. Also, they are the encounter incidents related crime, people who engage in violence or suspicious injuries frequently. So, documentation of patients’ condition came to the hospital and conservation of evidence are important in the inquiry of forensic medicine. The aim of the study was to determine the knowledge level of healthcare professional working at the emergency services regarding their approaches to common forensic cases. The study was comprised of 404 healthcare professional working (nurse, emergency medicine technician, health officer) at the emergency services of 6 state hospitals, 6 training and 6 research hospitals and 3 university hospitals in Ankara. Data was collected using questionnaire form which was developed by researches in the direction of literature. Questionnaire form is comprised of two sections. The first section includes 17 questions related demographic information about health care professional and 4 questions related Turkish laws. The second section includes 43 questions to the determination of knowledge level of health care professional’s working in the emergency department, about approaches to frequently encountered forensic cases. For the data evaluation of the study; Mann Whitney U test, Bonferroni correction Kruskal Wallis H test and Chi Square tests have been used. According to study, it’s said that there is no forensic medicine expert in the foundation by 73.4% of health care professionals. Two third (66%) of participants’ in emergency department reported daily average 7 or above forensic cases applied to the emergency department and 52.1% of participants did not evaluate incidents came to the emergency department as a forensic case. Most of the participants informed 'duty of preservation of evidence' is health care professionals duty related forensic cases. In result, we determinated that knowledge level of health care professional working in the emergency department, about approaches to frequently encountered forensic cases, is not the expected level. Because we found that most of them haven't received education about forensic nursing.Postgraduates participants, educated health professional about forensic nursing, staff who applied to sources about forensic nursing and staff who evaluated emergency department cases as forensic cases have significantly higher level of knowledge. Moreover, it’s found that forensic cases diagnosis score is the highest in health officer and university graduated. Health care professional’s deficiency in knowledge about forensic cases can cause defects in operation of the forensic process because of mistakes in collecting and conserving of evidence. It is obvious that training about the approach to forensic nursing should be arranged.

Keywords: emergency nurses, forensic case, forensic nursing, level of knowledge

Procedia PDF Downloads 295

Authors: Endalkachew Abebe Kebede, Bojin Bojinov, Andon Vasilev Andonov, Orhan Dengiz

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Remote sensing has a potential application in assessing and monitoring the plants' biophysical properties using the spectral responses of plants and soils within the electromagnetic spectrum. However, only a few reports compare the performance of different remote sensing sensors against in-situ field spectral measurement. The current study assessed the potential applications of open data source satellite images (Sentinel 2 and Landsat 9) in estimating the biophysical properties of the wheat crop on a study farm found in the village of OvchaMogila. A Landsat 9 (30 m resolution) and Sentinel-2 (10 m resolution) satellite images with less than 10% cloud cover have been extracted from the open data sources for the period of December 2021 to April 2022. An Unmanned Aerial Vehicle (UAV) has been used to capture the spectral response of plant leaves. In addition, SpectraVue 710s Leaf Spectrometer was used to measure the spectral response of the crop in April at five different locations within the same field. The ten most common vegetation indices have been selected and calculated based on the reflectance wavelength range of remote sensing tools used. The soil samples have been collected in eight different locations within the farm plot. The different physicochemical properties of the soil (pH, texture, N, P₂O₅, and K₂O) have been analyzed in the laboratory. The finer resolution images from the UAV and the Leaf Spectrometer have been used to validate the satellite images. The performance of different sensors has been compared based on the measured leaf spectral response and the extracted vegetation indices using the five sampling points. A scatter plot with the coefficient of determination (R2) and Root Mean Square Error (RMSE) and the correlation (r) matrix prepared using the corr and heatmap python libraries have been used for comparing the performance of Sentinel 2 and Landsat 9 VIs compared to the drone and SpectraVue 710s spectrophotometer. The soil analysis revealed the study farm plot is slightly alkaline (8.4 to 8.52). The soil texture of the study farm is dominantly Clay and Clay Loam.The vegetation indices (VIs) increased linearly with the growth of the plant. Both the scatter plot and the correlation matrix showed that Sentinel 2 vegetation indices have a relatively better correlation with the vegetation indices of the Buteo dronecompared to the Landsat 9. The Landsat 9 vegetation indices somewhat align better with the leaf spectrometer. Generally, the Sentinel 2 showed a better performance than the Landsat 9. Further study with enough field spectral sampling and repeated UAV imaging is required to improve the quality of the current study.

Keywords: landsat 9, leaf spectrometer, sentinel 2, UAV

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Authors: J. Manuel Bello-López, Julieta Rojo-Medina

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Introduction: The transplant of Umbilical Cord Blood Units (UCBU) are a therapeutic possibility for patients with oncohaematological disorders, especially in children. In Mexico, 48.5% of oncological diseases in children 1-4 years old are leukemias; whereas in patients 5-14 and 15-24 years old, lymphomas and leukemias represent the second and third cause of death in these groups respectively. Therefore it is necessary to have more registries of UCBU in order to ensure genetic diversity in the country; the above because the search for appropriate a UCBU is increasingly difficult for patients of mixed ethnicity. Objective: To estimate the genetic diversity (polymorphisms) of Human Leucocyte Antigen (HLA) Class I (A, B) and Class II (DRB1) in UCBU cryopreserved for transplant at Cord Blood Bank of the NCBT. Material and Methods: HLA typing of 533 UCBU for transplant was performed from 2003-2012 at the Histocompatibility Laboratory from the Research Department (evaluated by Los Angeles Ca. Immunogenetics Center) of the NCBT. Class I HLA-A, HLA-B and Class II HLA-DRB1 typing was performed using medium resolution Sequence-Specific Primer (SSP). In cases of an ambiguity detected by SSP; Sequence-Specific Oligonucleotide (SSO) method was carried out. A strict analysis of populations genetic parameters were done in 5 representative UCBU populations. Results: 46.5% of UCBU were collected from Mexico City, State of Mexico (30.95%), Puebla (8.06%), Morelos (6.37%) and Veracruz (3.37%). The remaining UCBU (4.75%) are represented by other states. The identified genotypes correspond to Amerindian origins (HLA-A*02, 31; HLA-B*39, 15, 48), Caucasian (HLA-A*02, 68, 01, 30, 31; HLA-B*35, 15, 40, 44, 07 y HLA-DRB1*04, 08, 07, 15, 03, 14), Oriental (HLA-A*02, 30, 01, 31; HLA-B* 35, 39, 15, 40, 44, 07,48 y HLA-DRB1*04, 07,15, 03) and African (HLA-A*30 y HLA-DRB1*03). The genetic distances obtained by Cavalli-Sforza analysis of the five states showed significant genetic differences by comparing genetic frequencies. The shortest genetic distance exists between Mexico City and the state of Puebla (0.0039) and the largest between Veracruz and Morelos (0.0084). In order to identify significant differences between this states, the ANOVA test was performed. This demonstrates that UCBU is significantly different according to their origin (P <0.05). This is shown by the divergence between arms at the Dendogram of Neighbor-Joining. Conclusions: The NCBT provides UCBU in patients with oncohaematological disorders in all the country. There is a group of patients for which not compatible UCBU can be find due to the mixed ethnic origin. For example, the population of northern Mexico is mostly Caucasian. Most of the NCBT donors are of various ethnic origins, predominantly Amerindians and Caucasians; although some ethnic minorities like Oriental, African and pure Indian ethnics are not represented. The NCBT is, therefore, establishing agreements with different states of Mexico to promote the altruistic donation of Umbilical Cord Blood in order to enrich the genetic diversity in its files.

Keywords: cord blood, genetic diversity, human leucocyte antigen, transplant

Procedia PDF Downloads 382

Authors: Ming-Yeh Lu, Shiao-Shing Chen, Saikat Sinha Ray, Hung-Te Hsu

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Recently, anaerobic membrane bioreactors (AnMBRs) has been widely utilized, which combines anaerobic biological treatment process and membrane filtration, that can be present an attractive option for wastewater treatment and water reuse. Conventional AnMBR is having several advantages, such as improving effluent quality, compact space usage, lower sludge yield, without aeration and production of energy. However, the removal of nitrogen and phosphorus in the AnMBR permeate was negligible which become the biggest disadvantage. In recent years, forward osmosis (FO) is an emerging technology that utilizes osmotic pressure as driving force to extract clean water without additional external pressure. The pore size of FO membrane is kindly mentioned the pore size, so nitrogen or phosphorus could effectively improve removal of nitrogen or phosphorus. Anaerobic bioreactor with FO membrane (AnOMBR) can retain the concentrate organic matters and nutrients. However, phosphorus is a non-renewable resource. Due to the high rejection property of FO membrane, the high amount of phosphorus could be recovered from the combination of AnMBR and FO. In this study, development of novel submerged anaerobic osmotic membrane bioreactor integrated with periodic microfiltration (MF) extraction for simultaneous phosphorus and clean water recovery from wastewater was evaluated. A laboratory-scale AnOMBR utilizes cellulose triacetate (CTA) membranes with effective membrane area of 130 cm² was fully submerged into a 5.5 L bioreactor at 30-35℃. Active layer-facing feed stream orientation was utilized, for minimizing fouling and scaling. Additionally, a peristaltic pump was used to circulate draw solution (DS) at a cross flow velocity of 0.7 cm/s. Magnesium sulphate (MgSO₄) solution was used as DS. Microfiltration membrane periodically extracted about 1 L solution when the TDS reaches to 5 g/L to recover phosphorus and simultaneous control the salt accumulation in the bioreactor. During experiment progressed, the average water flux was achieved around 1.6 LMH. The AnOMBR process show greater than 95% removal of soluble chemical oxygen demand (sCOD), nearly 100% of total phosphorous whereas only partial removal of ammonia, and finally average methane production of 0.22 L/g sCOD was obtained. Therefore, AnOMBR system periodically utilizes MF membrane extracted for phosphorus recovery with simultaneous pH adjustment. The overall performance demonstrates that a novel submerged AnOMBR system is having potential for simultaneous wastewater treatment and resource recovery from wastewater, and hence, the new concept of this system can be used to replace for conventional AnMBR in the future.

Keywords: anaerobic treatment, forward osmosis, phosphorus recovery, membrane bioreactor

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Authors: Gerald Amatre, Julius Bunny Lejju, Morgan Andama

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Despite several interventions, jigger flea infestations continue to be reported in the Busoga sub-region in Eastern Uganda. The purpose of this study was to identify factors that expose the indigenous people to jigger flea infestations and evaluate the effectiveness of any indigenous materials used in flea control by the affected communities. Flea compositions in residences were described, factors associated with flea infestation and indigenous materials used in flea control were evaluated. Field surveys were conducted in the affected communities after obtaining preliminary information on jigger infestation from the offices of the District Health Inspectors to identify the affected villages and households. Informed consent was then sought from the local authorities and household heads to conduct the study. Focus group discussions were conducted with key district informants, namely, the District Health Inspectors, District Entomologists and representatives from the District Health Office. A GPS coordinate was taken at central point at every household enrolled. Fleas were trapped inside residences using Kilonzo traps. A Kilonzo Trap comprised a shallow pan, about three centimetres deep, filled to the brim with water. The edges of the pan were smeared with Vaseline to prevent fleas from crawling out. Traps were placed in the evening and checked every morning the following day. The trapped fleas were collected in labelled vials filled with 70% aqueous ethanol and taken to the laboratory for identification. Socio-economic and environmental data were collected. The results indicate that the commonest flea trapped in the residences was the cat flea (Ctenocephalides felis) (50%), followed by Jigger flea (Tunga penetrans) (46%) and rat flea (Xenopsylla Cheopis) (4%), respectively. The average size of residences was seven squire metres with a mean of six occupants. The residences were generally untidy; with loose dusty floors and the brick walls were not plastered. The majority of the jigger affected households were headed by peasants (86.7%) and artisans (13.3%). The household heads mainly stopped at primary school level (80%) and few at secondary school level (20%). The jigger affected households were mainly headed by peasants of low socioeconomic status. The affected community members use soil-cow dung-ash mixture to smear floors of residences as the only measure to control fleas. This method was found to be ineffective in controlling the insects. The study recommends that home improvement campaigns be continued in the affected communities to improve sanitation and hygiene in residences as one of the interventions to combat flea infestations. Other cheap, available and effective means should be identified to curb jigger flea infestations.

Keywords: cow dung-soil-ash mixture, infestations, jigger flea, Tunga penetrans

Procedia PDF Downloads 136

Authors: Iris Vural Gursel, Jonathan Moncada, Ernst Worrell, Andrea Ramirez

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In order to achieve the transition from a fossil to bio-based economy, biomass needs to replace fossil resources in meeting the world’s energy and chemical needs. This calls for development of biorefinery systems allowing cost-efficient conversion of biomass to chemicals. In biorefinery systems, feedstock is converted to key intermediates called platforms which are converted to wide range of marketable products. The C6 sugars platform stands out due to its unique versatility as precursor for multiple valuable products. Among the different potential routes from C6 sugars to bio-based chemicals, 1,3-butadiene and ε-caprolactam appear to be of great interest. Butadiene is an important chemical for the production of synthetic rubbers, while caprolactam is used in production of nylon-6. In this study, ex-ante techno-economic performance of 1,3-butadiene and ε-caprolactam routes from C6 sugars were assessed. The aim is to provide insight from an early stage of development into the potential of these new technologies, and the bottlenecks and key cost-drivers. Two cases for each product line were analyzed to take into consideration the effect of possible changes on the overall performance of both butadiene and caprolactam production. Conceptual process design for the processes was developed using Aspen Plus based on currently available data from laboratory experiments. Then, operating and capital costs were estimated and an economic assessment was carried out using Net Present Value (NPV) as indicator. Finally, sensitivity analyses on processing capacity and prices was done to take into account possible variations. Results indicate that both processes perform similarly from an energy intensity point of view ranging between 34-50 MJ per kg of main product. However, in terms of processing yield (kg of product per kg of C6 sugar), caprolactam shows higher yield by a factor 1.6-3.6 compared to butadiene. For butadiene production, with the economic parameters used in this study, for both cases studied, a negative NPV (-642 and -647 M€) was attained indicating economic infeasibility. For the caprolactam production, one of the cases also showed economic infeasibility (-229 M€), but the case with the higher caprolactam yield resulted in a positive NPV (67 M€). Sensitivity analysis indicated that the economic performance of caprolactam production can be improved with the increase in capacity (higher C6 sugars intake) reflecting benefits of the economies of scale. Furthermore, humins valorization for heat and power production was considered and found to have a positive effect. Butadiene production was found sensitive to the price of feedstock C6 sugars and product butadiene. However, even at 100% variation of the two parameters, butadiene production remained economically infeasible. Overall, the caprolactam production line shows higher economic potential in comparison to that of butadiene. The results are useful in guiding experimental research and providing direction for further development of bio-based chemicals.

Keywords: bio-based chemicals, biorefinery, C6 sugars, economic analysis, process modelling

Procedia PDF Downloads 152

Authors: C. Miranda, S. Cunha, R. Soares, M. Maia, G. Igrejas, F. Silva, P. Poeta

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The worldwide inappropriate use of antibiotics has increased the emergence of antimicrobial-resistant microorganisms isolated from animals, humans, food, and the environment. To combat this complex and multifaceted problem is essential to know the prevalence in livestock animals and possible ways of transmission among animals and between these and humans. Enterococci species, in particular E. faecalis and E. faecium, are the most common nosocomial bacteria, causing infections in animals and humans. Thus, the aim of this study was to characterize resistance and virulence factors genes among two enterococci species isolated from calf colostrums in Portuguese dairy farms. The 55 enterococci isolates (44 E. faecalis and 11 E. faecium) were tested for the presence of the resistance genes for the following antibiotics: erythromicyn (ermA, ermB, and ermC), tetracycline (tetL, tetM, tetK, and tetO), quinupristin/dalfopristin (vatD and vatE) and vancomycin (vanB). Of which, 25 isolates (15 E. faecalis and 10 E. faecium) were tested until now for 8 virulence factors genes (esp, ace, gelE, agg, cpd, cylA, cylB, and cylLL). The resistance and virulence genes were performed by PCR, using specific primers and conditions. Negative and positive controls were used in all PCR assays. All enterococci isolates showed resistance to erythromicyn and tetracycline through the presence of the genes: ermB (n=29, 53%), ermC (n=10, 18%), tetL (n=49, 89%), tetM (n=39, 71%) and tetK (n=33, 60%). Only two (4%) E. faecalis isolates showed the presence of tetO gene. No resistance genes for vancomycin were found. The virulence genes detected in both species were cpd (n=17, 68%), agg (n=16, 64%), ace (n=15, 60%), esp (n=13, 52%), gelE (n=13, 52%) and cylLL (n=8, 32%). In general, each isolate showed at least three virulence genes. In three E. faecalis isolates was not found virulence genes and only E. faecalis isolates showed virulence genes for cylA (n=4, 16%) and cylB (n=6, 24%). In conclusion, these colostrum samples that were consumed by calves demonstrated the presence of antibiotic-resistant enterococci harbored virulence genes. This genotypic characterization is crucial to control the antibiotic-resistant bacteria through the implementation of restricts measures safeguarding public health. Acknowledgements: This work was funded by the R&D Project CAREBIO2 (Comparative assessment of antimicrobial resistance in environmental biofilms through proteomics - towards innovative theragnostic biomarkers), with reference NORTE-01-0145-FEDER-030101 and PTDC/SAU-INF/30101/2017, financed by the European Regional Development Fund (ERDF) through the Northern Regional Operational Program (NORTE 2020) and the Foundation for Science and Technology (FCT). This work was supported by the Associate Laboratory for Green Chemistry - LAQV which is financed by national funds from FCT/MCTES (UIDB/50006/2020 and UIDP/50006/2020).

Keywords: antimicrobial resistance, calf, colostrums, enterococci

Procedia PDF Downloads 200