Search results for: animal dunk
Commenced in January 2007
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Edition: International
Paper Count: 1139

Search results for: animal dunk

89 A Dynamic Model for Circularity Assessment of Nutrient Recovery from Domestic Sewage

Authors: Anurag Bhambhani, Jan Peter Van Der Hoek, Zoran Kapelan

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The food system depends on the availability of Phosphorus (P) and Nitrogen (N). Growing population, depleting Phosphorus reserves and energy-intensive industrial nitrogen fixation are threats to their future availability. Recovering P and N from domestic sewage water offers a solution. Recovered P and N can be applied to agricultural land, replacing virgin P and N. Thus, recovery from sewage water offers a solution befitting a circular economy. To ensure minimum waste and maximum resource efficiency a circularity assessment method is crucial to optimize nutrient flows and minimize losses. Material Circularity Indicator (MCI) is a useful method to quantify the circularity of materials. It was developed for materials that remain within the market and recently extended to include biotic materials that may be composted or used for energy recovery after end-of-use. However, MCI has not been used in the context of nutrient recovery. Besides, MCI is time-static, i.e., it cannot account for dynamic systems such as the terrestrial nutrient cycles. Nutrient application to agricultural land is a highly dynamic process wherein flows and stocks change with time. The rate of recycling of nutrients in nature can depend on numerous factors such as prevailing soil conditions, local hydrology, the presence of animals, etc. Therefore, a dynamic model of nutrient flows with indicators is needed for the circularity assessment. A simple substance flow model of P and N will be developed with the help of flow equations and transfer coefficients that incorporate the nutrient recovery step along with the agricultural application, the volatilization and leaching processes, plant uptake and subsequent animal and human uptake. The model is then used for calculating the proportions of linear and restorative flows (coming from reused/recycled sources). The model will simulate the adsorption process based on the quantity of adsorbent and nutrient concentration in the water. Thereafter, the application of the adsorbed nutrients to agricultural land will be simulated based on adsorbate release kinetics, local soil conditions, hydrology, vegetation, etc. Based on the model, the restorative nutrient flow (returning to the sewage plant following human consumption) will be calculated. The developed methodology will be applied to a case study of resource recovery from wastewater. In the aforementioned case study located in Italy, biochar or zeolite is to be used for recovery of P and N from domestic sewage through adsorption and thereafter, used as a slow-release fertilizer in agriculture. Using this model, information regarding the efficiency of nutrient recovery and application can be generated. This can help to optimize the recovery process and application of the nutrients. Consequently, this will help to optimize nutrient recovery and application and reduce the dependence of the food system on the virgin extraction of P and N.

Keywords: circular economy, dynamic substance flow, nutrient cycles, resource recovery from water

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88 The Improved Therapeutic Effect of Trans-Cinnamaldehyde on Adipose-Derived Stem Cells without Chemical Induction

Authors: Karthyayani Rajamani, Yi-Chun Lin, Tung-Chou Wen, Jeanne Hsieh, Yi-Maun Subeq, Jen-Wei Liu, Po-Cheng Lin, Horng-Jyh Harn, Shinn-Zong Lin, Tzyy-Wen Chiou

Abstract:

Assuring cell quality is an essential parameter for the success of stem cell therapy, utilization of various components to improve this potential has been the primary goal of stem cell research. The aim of this study was not only to demonstrate the capacity of trans-cinnamaldehyde (TC) to reverse stress-induced senescence but also improve the therapeutic abilities of stem cells. Because of the availability and the promising application potential in regenerative medicine, adipose-derived stem cells (ADSCs) were chosen for the study. We found that H2O2 treatment resulted in the expression of senescence characteristics in the ADSCs, including decreased proliferation rate, increased senescence-associated- β-galactosidase (SA-β-gal) activity, decreased SIRT1 (silent mating type information regulation 2 homologs) expression and decreased telomerase activity. However, TC treatment was sufficient to rescue or reduce the effects of H2O2 induction, ultimately leading to an increased proliferation rate, a decrease in the percentage of SA-β-gal positive cells, upregulation of SIRT1 expression, and increased telomerase activity of the senescent ADSCs at the cellular level. Further recently it was observed that the ADSCs were treated with TC without induction of senescence, all the before said positives were observed. Moreover, a chemically induced liver fibrosis animal model was used to evaluate the functionality of these rescued cells in vivo. Liver dysfunction was established by injecting 200 mg/kg thioacetamide (TAA) intraperitoneally into Wistar rats every third day for 60 days. The experimental rats were separated into groups; normal group (rats without TAA induction), sham group (without ADSC transplantation), positive control group (transplanted with normal ADSCs); H2O2 group (transplanted with H2O2 -induced senescent ADSCs), H2O2+TC group (transplanted with ADSCs pretreated with H2O2 and then further treated with TC) and TC group (ADSC treated with TC without H2O2 treatment). In the transplantation group, 1 × 106 human ADSCs were introduced into each rat via direct liver injection. Based on the biochemical analysis and immunohistochemical staining results, it was determined that the therapeutic effects on liver fibrosis by the induced senescent ADSCs (H2O2 group) were not as significant as those exerted by the normal ADSCs (the positive control group). However, the H2O2+TC group showed significant reversal of liver damage when compared to the H2O2 group 1 week post-transplantation. Further ADSCs without H2O2 treatment but with just TC treatment performed much better than all the groups. These data confirmed that the TC treatment had the potential to improve the therapeutic effect of ADSCs. It is therefore suggested that TC has potential applications in maintaining stem cell quality and could possibly aid in the treatment of senescence-related disorders.

Keywords: senescence, SIRT1, adipose derived stem cells, liver fibrosis

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87 Development of a One Health and Comparative Medicine Curriculum for Medical Students

Authors: Aliya Moreira, Blake Duffy, Sam Kosinski, Kate Heckman, Erika Steensma

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Introduction: The One Health initiative promotes recognition of the interrelatedness between people, animals, plants, and their shared environment. The field of comparative medicine studies the similarities and differences between humans and animals for the purpose of advancing medical sciences. Currently, medical school education is narrowly focused on human anatomy and physiology, but as the COVID-19 pandemic has demonstrated, a holistic understanding of health requires comprehension of the interconnection between health and the lived environment. To prepare future physicians for unique challenges from emerging zoonoses to climate change, medical students can benefit from exposure to and experience with One Health and Comparative Medicine content. Methods: In January 2020, an elective course for medical students on One Health and Comparative Medicine was created to provide medical students with the background knowledge necessary to understand the applicability of animal and environmental health in medical research and practice. The 2-week course was continued in January 2021, with didactic and experiential activities taking place virtually due to the COVID-19 pandemic. In response to student feedback, lectures were added to expand instructional content on zoonotic and wildlife diseases for the second iteration of the course. Other didactic sessions included interprofessional lectures from 20 physicians, veterinarians, public health professionals, and basic science researchers. The first two cohorts of students were surveyed regarding One Health and Comparative Medicine concepts at the beginning and conclusion of the course. Results: 16 medical students have completed the comparative medicine course thus far, with 87.5% (n=14) completing pre-and post-course evaluations. 100% of student respondents indicated little to no exposure to comparative medicine or One Health concepts during medical school. Following the course, 100% of students felt familiar or very familiar with comparative medicine and One Health concepts. To assess course efficacy, questions were evaluated on a five-point Likert scale. 100% agreed or strongly agreed that learning Comparative Medicine and One Health topics augmented their medical education. 100% agreed or strongly agreed that a course covering this content should be regularly offered to medical students. Conclusions: Data from the student evaluation surveys demonstrate that the Comparative Medicine course was successful in increasing medical student knowledge of Comparative Medicine and One Health. Results also suggest that interprofessional training in One Health and Comparative Medicine is applicable and useful for medical trainees. Future iterations of this course could capitalize on the inherently interdisciplinary nature of these topics by enrolling students from veterinary and public health schools into a longitudinal course. Such recruitment may increase the course’s value by offering multidisciplinary student teams the opportunity to conduct research projects, thereby strengthening both the individual learning experience as well as sparking future interprofessional research ventures. Overall, these efforts to educate medical students in One Health topics should be reproducible at other institutions, preparing more future physicians for the diverse challenges they will encounter in practice.

Keywords: medical education, interprofessional instruction, one health, comparative medicine

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86 Insights on the Halal Status of Antineoplastic and Immunomodulating Agents and Nutritional and Dietary Supplements in Malaysia

Authors: Suraiya Abdul Rahman, Perasna M. Varma, Amrahi Buang, Zhari Ismail, Wan Rosalina W. Rosli, Ahmad Rashidi M. Tahir

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Background: Muslims has the obligation to ensure that everything they consume including medicines should be halal. With the growing demands for halal medicines in October 2012, Malaysia has launched the world's first Halal pharmaceutical standards called Malaysian Standard MS 2424:2012 Halal Pharmaceuticals-General Guidelines to serve as a basic requirement for halal pharmaceuticals in Malaysia. However, the biggest challenge faced by pharmaceutical companies to comply is finding the origin or source of the ingredients and determine their halal status. Aim: This study aims to determine the halal status of the antineoplastic and immunomodulating agents, and nutritional and dietary supplements by analysing the origin of their active pharmaceutical ingredients (API) and excipients to provide an insight on the common source and halal status of pharmaceutical ingredients and an indication on adjustment required in order to be halal compliance. Method: The ingredients of each product available in a government hospital in central of Malaysia and their sources were determined from the product package leaflets, information obtained from manufacturer, reliable websites and standard pharmaceutical references. The ingredients were categorised as halal, musbooh or haram based on the definition set in MS2424. Results: There were 162 medications included in the study where 123 (76%) were under the antineoplastic and immunomodulating agents group, while 39 (24%) were nutritional and dietary supplements. In terms of the medication halal status, the proportion of halal, musbooh and haram were 40.1% (n=65), 58.6% (n=95) and 1.2% (n=2) respectively. With regards to the API, there were 89 (52%) different active ingredient identified for antineoplastic and immunomodulating agents with the proportion of 89.9% (n=80) halal and 10.1% (n=9) were mushbooh. There were 83 (48%) active ingredient from the nutritional and dietary supplements group with proportion of halal and masbooh were 89.2% (n=74) and 10.8% (n=9) respectively. No haram APIs were identified in all therapeutic classes. There were a total of 176 excipients identified from the products ranges. It was found that majority of excipients are halal with the proportion of halal, masbooh and haram were at 82.4% (n=145), 17% (n=30) and 0.6% (n=1) respectively. With regards of the sources of the excipeints, most of masbooh excipients (76.7%, n = 23) were classified as masbooh because they have multiple possible origin which consist of animals, plant or others. The remaining 13.3% and 10% were classified as masbooh due to their ethanol and land animal origin respectively. The one haram excipient was gelatine of bovine-porcine origin. Masbooh ingredients found in this research were glycerol, tallow, lactose, polysorbate, dibasic sodium phosphate, stearic acid and magnesium stearate. Ethanol, gelatine, glycerol and magnesium stearate were the most common ingredients classified as mushbooh. Conclusion: This study shows that most API and excipients are halal. However the majority of the medicines in these products categories are mushbooh due to certain excipients only, which could be replaced with halal alternative excipients. This insight should encourage the pharmaceutical products manufacturers to go for halal certification to meet the increasing demand for Halal certified medications for the benefit of mankind.

Keywords: antineoplastic and immunomodulation agents, halal pharmaceutical, MS2424, nutritional and dietary supplements

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85 Analysis of Lesotho Wool Production and Quality Trends 2008-2018

Authors: Papali Maqalika

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Lesotho farmers produce significant quantities of Merino wool of a quality competitive on the global market and make a substantial impact on the economy of Lesotho. However, even with the economic contribution, the production and quality information and trends of this fibre has been recognised nor documented. This is a sombre shortcoming as Lesotho wool is unknown on international markets. The situation is worsened by the fact that Lesotho wool is auction together with South African wool, trading and benchmarking Lesotho wool are difficult not to mention attempts to advance its production and quality. Based on the information above, available data on Lesotho wool for 10 years were collected and analysed for trends to used in benchmarking where applicable. The fibre properties analysed include fibre diameter (fineness), vegetable matter and yield, application and price. These were selected because they are fundamental in determining fibre quality and price. Production of wool in Lesotho has increased slightly over the ten years covered by this study. It also became apparent that production and quality trends of Lesotho wool are greatly influenced by the farming practices, breed of sheep and climatic conditions. Greater adoption of the merino sheep breed, sheds/barns and sheep coats are suggested as ways to reduce mortality rate (due to extremely cold temperatures), to reduce the vegetable matter on the fibre thus improving the quality and increase yield per sheep and production as a whole. Some farming practices such as the lack of barns, supplementary feeding and veterinary care present constraints in wool production. The districts in the Highlands region were found to have the highest production of mostly wool, this being ascribed to better pastures, climatic, social and other conditions conducive to wool production. The production of Lesotho wool and its quality can be improved further, possibly because of the interventions the Ministry of Agriculture introduced through the Small Agricultural and Development Project (SADP) and other appropriate initiatives by the National Wool and Mohair Growers Association (NWMGA). The challenge however, remains the lack of direct involvement of the wool growers (farmers) in decisions making and policy development, this potentially influences and may lead to the reluctance to adopt the strategies. In some cases, the wool growers do not receive the benefits associated with the interventions immediately. Based on these discoveries; it is recommended that the relevant educators and researchers in wool and textile science, as well as the local wool farmers in Lesotho, be represented in policy and other decision making forums relating to these interventions. In this way, educational campaigns and training workshops will be demand driven with a better chance of adoption and success. This is because the direct beneficiaries will have been involved at inception and they will have a sense of ownership as well as intent to see them through successfully.

Keywords: lesotho wool, wool quality, wool production, lesotho economy, global market, apparel wool, database, textile science, exports, animal farming practices, intimate apparel, interventions

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84 Socio-Economic Transformation of Barpak Post-Earthquake Reconstruction

Authors: Sudikshya Bhandari, Jonathan K. London

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The earthquake of April 2015 was one of the biggest disasters in the history of Nepal. The epicenter was located near Barpak, north of the Gorkha district. Before the disaster, this settlement was a compact and homogeneous settlement manifesting its uniqueness through the social and cultural activities, and a distinct vernacular architecture. Narrow alleys with stone paved streets, buildings with slate roofs, and common spaces between the houses made this settlement socially, culturally, and environmentally cohesive. With the presence of micro hydro power plants, local economic activities enabled the local community to exist and thrive. Agriculture and animal rearing are the sources of livelihood for the majority of families, along with the booming homestays (where local people welcome guests to their home, as a business) and local shops. Most of these activities are difficult to find as the houses have been destroyed with the earthquake and the process of reconstruction has been transforming the outlook of the settlement. This study characterized the drastic transformation in Barpak post-earthquake, and analyzed the consequences of the reconstruction process. In addition, it contributes to comprehending a broader representation about unsustainability created by the lack of contextual post-disaster development. Since the research is based in a specific area, a case study approach was used. Sample houses were selected on the basis of ethnicity and house typology. Mixed methods such as key informant and semi structured interviews, focus groups, observations and photographs are used for the collection of data. The research focus is predominantly on the physical change of the house typology from vernacular to externally adopted designs. This transformation of the house entails socio-cultural changes such as social fragmentation with differences among the rich and the poor and decreases in the social connectivity within families and neighborhood. Families have found that new houses require more maintenance and resources that have increased their economic expenses. The study also found that the reconstructed houses are not thermally comfortable in the cold climate of Barpak, leading to the increased use of different sources of heating like electric heaters and more firewood. Lack of storage spaces for crops and livestock have discouraged them to pursue traditional means of livelihood and depend more on buying food from stores, ultimately making it less economical for most of the families. The transformation of space leading to the economic, social and cultural changes demonstrates the unsustainability of Barpak. Conclusions from the study suggest place based and inclusive planning and policy formations that include locals as partners, identifying the possible ways to minimize the impact and implement these recommendations into the future policy and planning scenarios.

Keywords: earthquake, Nepal, reconstruction, settlement, transformation

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83 Cytochrome B Diversity and Phylogeny of Egyptian Sheep Breeds

Authors: Othman E. Othman, Agnés Germot, Daniel Petit, Abderrahman Maftah

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Threats to the biodiversity are increasing due to the loss of genetic diversity within the species utilized in agriculture. Due to the progressive substitution of the less productive, locally adapted and native breeds by highly productive breeds, the number of threatened breeds is increased. In these conditions, it is more strategically important than ever to preserve as much the farm animal diversity as possible, to ensure a prompt and proper response to the needs of future generations. Mitochondrial (mtDNA) sequencing has been used to explain the origins of many modern domestic livestock species. Studies based on sequencing of sheep mitochondrial DNA showed that there are five maternal lineages in the world for domestic sheep breeds; A, B, C, D and E. Because of the eastern location of Egypt in the Mediterranean basin and the presence of fat-tailed sheep breeds- character quite common in Turkey and Syria- where genotypes that seem quite primitive, the phylogenetic studies of Egyptian sheep breeds become particularly attractive. We aimed in this work to clarify the genetic affinities, biodiversity and phylogeny of five Egyptian sheep breeds using cytochrome B sequencing. Blood samples were collected from 63 animals belonging to the five tested breeds; Barki, Rahmani, Ossimi, Saidi and Sohagi. The total DNA was extracted and the specific primer allowed the conventional PCR amplification of the cytochrome B region of mtDNA (approximately 1272 bp). PCR amplified products were purified and sequenced. The alignment of Sixty-three samples was done using BioEdit software. DnaSP 5.00 software was used to identify the sequence variation and polymorphic sites in the aligned sequences. The result showed that the presence of 34 polymorphic sites leading to the formation of 18 haplotypes. The haplotype diversity in five tested breeds ranged from 0.676 in Rahmani breed to 0.894 in Sohagi breed. The genetic distances (D) and the average number of pairwise differences (Dxy) between breeds were estimated. The lowest distance was observed between Rahmani and Saidi (D: 1.674 and Dxy: 0.00150) while the highest distance was observed between Ossimi and Sohagi (D: 5.233 and Dxy: 0.00475). Neighbour-joining (Phylogeny) tree was constructed using Mega 5.0 software. The sequences of the 63 analyzed samples were aligned with references sequences of different haplogroups. The phylogeny result showed the presence of three haplogroups (HapA, HapB and HapC) in the 63 examined samples. The other two haplogroups described in literature (HapD and HapE) were not found. The result showed that 50 out of 63 tested animals cluster with haplogroup B (79.37%) whereas 7 tested animals cluster with haplogroup A (11.11%) and 6 animals cluster with haplogroup C (9.52%). In conclusion, the phylogenetic reconstructions showed that the majority of Egyptian sheep breeds belonging to haplogroup B which is the dominant haplogroup in Eastern Mediterranean countries like Syria and Turkey. Some individuals are belonging to haplogroups A and C, suggesting that the crosses were done with other breeds for characteristic selection for growth and wool quality.

Keywords: cytochrome B, diversity, phylogheny, Egyptian sheep breeds

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82 Application of IoTs Based Multi-Level Air Quality Sensing for Advancing Environmental Monitoring in Pingtung County

Authors: Men An Pan, Hong Ren Chen, Chih Heng Shih, Hsing Yuan Yen

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Pingtung County is located in the southernmost region of Taiwan. During the winter season, pollutants due to insufficient dispersion caused by the downwash of the northeast monsoon lead to the poor air quality of the County. Through the implementation of various control methods, including the application of permits of air pollution, fee collection of air pollution, control oil fume of catering sectors, smoke detection of diesel vehicles, regular inspection of locomotives, and subsidies for low-polluting vehicles. Moreover, to further mitigate the air pollution, additional alternative controlling strategies are also carried out, such as construction site control, prohibition of open-air agricultural waste burning, improvement of river dust, and strengthening of road cleaning operations. The combined efforts have significantly reduced air pollutants in the County. However, in order to effectively and promptly monitor the ambient air quality, the County has subsequently deployed micro-sensors, with a total of 400 IoTs (Internet of Things) micro-sensors for PM2.5 and VOC detection and 3 air quality monitoring stations of the Environmental Protection Agency (EPA), covering 33 townships of the County. The covered area has more than 1,300 listed factories and 5 major industrial parks; thus forming an Internet of Things (IoTs) based multi-level air quality monitoring system. The results demonstrate that the IoTs multi-level air quality sensors combined with other strategies such as “sand and gravel dredging area technology monitoring”, “banning open burning”, “intelligent management of construction sites”, “real-time notification of activation response”, “nighthawk early bird plan with micro-sensors”, “unmanned aircraft (UAV) combined with land and air to monitor abnormal emissions”, and “animal husbandry odour detection service” etc. The satisfaction improvement rate of air control, through a 2021 public survey, reached a high percentage of 81%, an increase of 46% as compared to 2018. For the air pollution complaints for the whole year of 2021, the total number was 4213 in contrast to 7088 in 2020, a reduction rate reached almost 41%. Because of the spatial-temporal features of the air quality monitoring IoTs system by the application of microsensors, the system does assist and strengthen the effectiveness of the existing air quality monitoring network of the EPA and can provide real-time control of the air quality. Therefore, the hot spots and potential pollution locations can be timely determined for law enforcement. Hence, remarkable results were obtained for the two years. That is, both reduction of public complaints and better air quality are successfully achieved through the implementation of the present IoTs system for real-time air quality monitoring throughout Pingtung County.

Keywords: IoT, PM, air quality sensor, air pollution, environmental monitoring

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81 Variability and Stability of Bread and Durum Wheat for Phytic Acid Content

Authors: Gordana Branković, Vesna Dragičević, Dejan Dodig, Desimir Knežević, Srbislav Denčić, Gordana Šurlan-Momirović

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Phytic acid is a major pool in the flux of phosphorus through agroecosystems and represents a sum equivalent to > 50% of all phosphorus fertilizer used annually. Nutrition rich in phytic acid can substantially decrease micronutrients apsorption as calcium, zink, iron, manganese, copper due to phytate salts excretion by human and non-ruminant animals as poultry, swine and fish, having in common very scarce phytase activity, and consequently the ability to digest and utilize phytic acid, thus phytic acid derived phosphorus in animal waste contributes to water pollution. The tested accessions consisted of 15 genotypes of bread wheat (Triticum aestivum L. ssp. vulgare) and of 15 genotypes of durum wheat (Triticum durum Desf.). The trials were sown at the three test sites in Serbia: Rimski Šančevi (RS) (45º19´51´´N; 19º50´59´´E), Zemun Polje (ZP) (44º52´N; 20º19´E) and Padinska Skela (PS) (44º57´N 20º26´E) during two vegetation seasons 2010-2011 and 2011-2012. The experimental design was randomized complete block design with four replications. The elementary plot consisted of 3 internal rows of 0.6 m2 area (3 × 0.2 m × 1 m). Grains were grinded with Laboratory Mill 120 Perten (“Perten”, Sweden) (particles size < 500 μm) and flour was used for the analysis. Phytic acid grain content was determined spectrophotometrically with the Shimadzu UV-1601 spectrophotometer (Shimadzu Corporation, Japan). Objectives of this study were to determine: i) variability and stability of the phytic acid content among selected genotypes of bread and durum wheat, ii) predominant source of variation regarding genotype (G), environment (E) and genotype × environment interaction (GEI) from the multi-environment trial, iii) influence of climatic variables on the GEI for the phytic acid content. Based on the analysis of variance it had been determined that the variation of phytic acid content was predominantly influenced by environment in durum wheat, while the GEI prevailed for the variation of the phytic acid content in bread wheat. Phytic acid content expressed on the dry mass basis was in the range 14.21-17.86 mg g-1 with the average of 16.05 mg g-1 for bread wheat and 14.63-16.78 mg g-1 with the average of 15.91 mg g-1 for durum wheat. Average-environment coordination view of the genotype by environment (GGE) biplot was used for the selection of the most desirable genotypes for breeding for low phytic acid content in the sense of good stability and lower level of phytic acid content. The most desirable genotypes of bread and durum wheat for breeding for phytic acid were Apache and 37EDUYT /07 No. 7849. Models of climatic factors in the highest percentage (> 91%) were useful in interpreting GEI for phytic acid content, and included relative humidity in June, sunshine hours in April, mean temperature in April and winter moisture reserves for genotypes of bread wheat, as well as precipitation in June and April, maximum temperature in April and mean temperature in June for genotypes of durum wheat.

Keywords: genotype × environment interaction, phytic acid, stability, variability

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80 Interplay of Material and Cycle Design in a Vacuum-Temperature Swing Adsorption Process for Biogas Upgrading

Authors: Federico Capra, Emanuele Martelli, Matteo Gazzani, Marco Mazzotti, Maurizio Notaro

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Natural gas is a major energy source in the current global economy, contributing to roughly 21% of the total primary energy consumption. Production of natural gas starting from renewable energy sources is key to limit the related CO2 emissions, especially for those sectors that heavily rely on natural gas use. In this context, biomethane produced via biogas upgrading represents a good candidate for partial substitution of fossil natural gas. The upgrading process of biogas to biomethane consists in (i) the removal of pollutants and impurities (e.g. H2S, siloxanes, ammonia, water), and (ii) the separation of carbon dioxide from methane. Focusing on the CO2 removal process, several technologies can be considered: chemical or physical absorption with solvents (e.g. water, amines), membranes, adsorption-based systems (PSA). However, none emerged as the leading technology, because of (i) the heterogeneity in plant size, ii) the heterogeneity in biogas composition, which is strongly related to the feedstock type (animal manure, sewage treatment, landfill products), (iii) the case-sensitive optimal tradeoff between purity and recovery of biomethane, and iv) the destination of the produced biomethane (grid injection, CHP applications, transportation sector). With this contribution, we explore the use of a technology for biogas upgrading and we compare the resulting performance with benchmark technologies. The proposed technology makes use of a chemical sorbent, which is engineered by RSE and consists of Di-Ethanol-Amine deposited on a solid support made of γ-Alumina, to chemically adsorb the CO2 contained in the gas. The material is packed into fixed beds that cyclically undergo adsorption and regeneration steps. CO2 is adsorbed at low temperature and ambient pressure (or slightly above) while the regeneration is carried out by pulling vacuum and increasing the temperature of the bed (vacuum-temperature swing adsorption - VTSA). Dynamic adsorption tests were performed by RSE and were used to tune the mathematical model of the process, including material and transport parameters (i.e. Langmuir isotherms data and heat and mass transport). Based on this set of data, an optimal VTSA cycle was designed. The results enabled a better understanding of the interplay between material and cycle tuning. As exemplary application, the upgrading of biogas for grid injection, produced by an anaerobic digester (60-70% CO2, 30-40% CH4), for an equivalent size of 1 MWel was selected. A plant configuration is proposed to maximize heat recovery and minimize the energy consumption of the process. The resulting performances are very promising compared to benchmark solutions, which make the VTSA configuration a valuable alternative for biomethane production starting from biogas.

Keywords: biogas upgrading, biogas upgrading energetic cost, CO2 adsorption, VTSA process modelling

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79 LaeA/1-Velvet Interplay in Aspergillus and Trichoderma: Regulation of Secondary Metabolites and Cellulases

Authors: Razieh Karimi Aghcheh, Christian Kubicek, Joseph Strauss, Gerhard Braus

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Filamentous fungi are of considerable economic and social significance for human health, nutrition and in white biotechnology. These organisms are dominant producers of a range of primary metabolites such as citric acid, microbial lipids (biodiesel) and higher unsaturated fatty acids (HUFAs). In particular, they produce also important but structurally complex secondary metabolites with enormous therapeutic applications in pharmaceutical industry, for example: cephalosporin, penicillin, taxol, zeranol and ergot alkaloids. Several fungal secondary metabolites, which are significantly relevant to human health do not only include antibiotics, but also e.g. lovastatin, a well-known antihypercholesterolemic agent produced by Aspergillus. terreus, or aflatoxin, a carcinogen produced by A. flavus. In addition to their roles for human health and agriculture, some fungi are industrially and commercially important: Species of the ascomycete genus Hypocrea spp. (teleomorph of Trichoderma) have been demonstrated as efficient producer of highly active cellulolytic enzymes. This trait makes them effective in disrupting and depolymerization of lignocellulosic materials and thus applicable tools in number of biotechnological areas as diverse as clothes-washing detergent, animal feed, and pulp and fuel productions. Fungal LaeA/LAE1 (Loss of aflR Expression A) homologs their gene products act at the interphase between secondary metabolisms, cellulase production and development. Lack of the corresponding genes results in significant physiological changes including loss of secondary metabolite and lignocellulose degrading enzymes production. At the molecular level, the encoded proteins are presumably methyltransferases or demethylases which act directly or indirectly at heterochromatin and interact with velvet domain proteins. Velvet proteins bind to DNA and affect expression of secondary metabolites (SMs) genes and cellulases. The dynamic interplay between LaeA/LAE1, velvet proteins and additional interaction partners is the key for an understanding of the coordination of metabolic and morphological functions of fungi and is required for a biotechnological control of the formation of desired bioactive products. Aspergilli and Trichoderma represent different biotechnologically significant species with significant differences in the LaeA/LAE1-Velvet protein machinery and their target proteins. We, therefore, performed a comparative study of the interaction partners of this machinery and the dynamics of the various protein-protein interactions using our robust proteomic and mass spectrometry techniques. This enhances our knowledge about the fungal coordination of secondary metabolism, cellulase production and development and thereby will certainly improve recombinant fungal strain construction for the production of industrial secondary metabolite or lignocellulose hydrolytic enzymes.

Keywords: cellulases, LaeA/1, proteomics, secondary metabolites

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78 Viscoelastic Behavior of Human Bone Tissue under Nanoindentation Tests

Authors: Anna Makuch, Grzegorz Kokot, Konstanty Skalski, Jakub Banczorowski

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Cancellous bone is a porous composite of a hierarchical structure and anisotropic properties. The biological tissue is considered to be a viscoelastic material, but many studies based on a nanoindentation method have focused on their elasticity and microhardness. However, the response of many organic materials depends not only on the load magnitude, but also on its duration and time course. Depth Sensing Indentation (DSI) technique has been used for examination of creep in polymers, metals and composites. In the indentation tests on biological samples, the mechanical properties are most frequently determined for animal tissues (of an ox, a monkey, a pig, a rat, a mouse, a bovine). However, there are rare reports of studies of the bone viscoelastic properties on microstructural level. Various rheological models were used to describe the viscoelastic behaviours of bone, identified in the indentation process (e. g Burgers model, linear model, two-dashpot Kelvin model, Maxwell-Voigt model). The goal of the study was to determine the influence of creep effect on the mechanical properties of human cancellous bone in indentation tests. The aim of this research was also the assessment of the material properties of bone structures, having in mind the energy aspects of the curve (penetrator loading-depth) obtained in the loading/unloading cycle. There was considered how the different holding times affected the results within trabecular bone.As a result, indentation creep (CIT), hardness (HM, HIT, HV) and elasticity are obtained. Human trabecular bone samples (n=21; mean age 63±15yrs) from the femoral heads replaced during hip alloplasty were removed and drained from alcohol of 1h before the experiment. The indentation process was conducted using CSM Microhardness Tester equipped with Vickers indenter. Each sample was indented 35 times (7 times for 5 different hold times: t1=0.1s, t2=1s, t3=10s, t4=100s and t5=1000s). The indenter was advanced at a rate of 10mN/s to 500mN. There was used Oliver-Pharr method in calculation process. The increase of hold time is associated with the decrease of hardness parameters (HIT(t1)=418±34 MPa, HIT(t2)=390±50 MPa, HIT(t3)= 313±54 MPa, HIT(t4)=305±54 MPa, HIT(t5)=276±90 MPa) and elasticity (EIT(t1)=7.7±1.2 GPa, EIT(t2)=8.0±1.5 GPa, EIT(t3)=7.0±0.9 GPa, EIT(t4)=7.2±0.9 GPa, EIT(t5)=6.2±1.8 GPa) as well as with the increase of the elastic (Welastic(t1)=4.11∙10-7±4.2∙10-8Nm, Welastic(t2)= 4.12∙10-7±6.4∙10-8 Nm, Welastic(t3)=4.71∙10-7±6.0∙10-9 Nm, Welastic(t4)= 4.33∙10-7±5.5∙10-9Nm, Welastic(t5)=5.11∙10-7±7.4∙10-8Nm) and inelastic (Winelastic(t1)=1.05∙10-6±1.2∙10-7 Nm, Winelastic(t2) =1.07∙10-6±7.6∙10-8 Nm, Winelastic(t3)=1.26∙10-6±1.9∙10-7Nm, Winelastic(t4)=1.56∙10-6± 1.9∙10-7 Nm, Winelastic(t5)=1.67∙10-6±2.6∙10-7)) reaction of materials. The indentation creep increased logarithmically (R2=0.901) with increasing hold time: CIT(t1) = 0.08±0.01%, CIT(t2) = 0.7±0.1%, CIT(t3) = 3.7±0.3%, CIT(t4) = 12.2±1.5%, CIT(t5) = 13.5±3.8%. The pronounced impact of creep effect on the mechanical properties of human cancellous bone was observed in experimental studies. While the description elastic-inelastic, and thus the Oliver-Pharr method for data analysis, may apply in few limited cases, most biological tissues do not exhibit elastic-inelastic indentation responses. Viscoelastic properties of tissues may play a significant role in remodelling. The aspect is still under an analysis and numerical simulations. Acknowledgements: The presented results are part of the research project founded by National Science Centre (NCN), Poland, no.2014/15/B/ST7/03244.

Keywords: bone, creep, indentation, mechanical properties

Procedia PDF Downloads 172
77 Technology Assessment of the Collection of Cast Seaweed and Use as Feedstock for Biogas Production- The Case of SolrøD, Denmark

Authors: Rikke Lybæk, Tyge Kjær

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The Baltic Sea is suffering from nitrogen and phosphorus pollution, which causes eutrophication of the maritime environment and hence threatens the biodiversity of the Baltic Sea area. The intensified quantity of nutrients in the water has created challenges with the growth of seaweed being discarded on beaches around the sea. The cast seaweed has led to odor problems hampering the use of beach areas around the Bay of Køge in Denmark. This is the case in, e.g., Solrød Municipality, where recreational activities have been disrupted when cast seaweed pile up on the beach. Initiatives have, however, been introduced within the municipality to remove the cast seaweed from the beach and utilize it for renewable energy production at the nearby Solrød Biogas Plant, thus being co-digested with animal manure for power and heat production. This paper investigates which type of technology application’s have been applied in the effort to optimize the collection of cast seaweed, and will further reveal, how the seaweed has been pre-treated at the biogas plant to be utilized for energy production the most efficient, hereunder the challenges connected with the content of sand. Heavy metal contents in the seaweed and how it is managed will also be addressed, which is vital as the digestate is utilized as soil fertilizer on nearby farms. Finally, the paper will outline the energy production scheme connected to the use of seaweed as feedstock for biogas production, as well as the amount of nitrogen-rich fertilizer produced. The theoretical approach adopted in the paper relies on the thinking of Circular Bio-Economy, where biological materials are cascaded and re-circulated etc., to increase and extend their value and usability. The data for this research is collected as part of the EU Interreg project “Cluster On Anaerobic digestion, environmental Services, and nuTrients removAL” (COASTAL Biogas), 2014-2020. Data gathering consists of, e.g., interviews with relevant stakeholders connected to seaweed collection and operation of the biogas plant in Solrød Municipality. It further entails studies of progress and evaluation reports from the municipality, analysis of seaweed digestion results from scholars connected to the research, as well as studies of scientific literature to supplement the above. Besides this, observations and photo documentation have been applied in the field. This paper concludes, among others, that the seaweed harvester technology currently adopted is functional in the maritime environment close to the beachfront but inadequate in collecting seaweed directly on the beach. New technology hence needs to be developed to increase the efficiency of seaweed collection. It is further concluded that the amount of sand transported to Solrød Biogas Plant with the seaweed continues to pose challenges. The seaweed is pre-treated for sand in a receiving tank with a strong stirrer, washing off the sand, which ends at the bottom of the tank where collected. The seaweed is then chopped by a macerator and mixed with the other feedstock. The wear down of the receiving tank stirrer and the chopper are, however, significant, and new methods should be adopted.

Keywords: biogas, circular bio-economy, Denmark, maritime technology, cast seaweed, solrød municipality

Procedia PDF Downloads 293
76 Influence of Cryo-Grinding on Particle Size Distribution of Proso Millet Bran Fraction

Authors: Maja Benkovic, Dubravka Novotni, Bojana Voucko, Duska Curic, Damir Jezek, Nikolina Cukelj

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Cryo-grinding is an ultra-fine grinding method used in the pharmaceutical industry, production of herbs and spices and in the production and handling of cereals, due to its ability to produce powders with small particle sizes which maintain their favorable bioactive profile. The aim of this study was to determine the particle size distributions of the proso millet (Panicum miliaceum) bran fraction grinded at cryogenic temperature (using liquid nitrogen (LN₂) cooling, T = - 196 °C), in comparison to non-cooled grinding. Proso millet bran is primarily used as an animal feed, but has a potential in food applications, either as a substrate for extraction of bioactive compounds or raw material in the bakery industry. For both applications finer particle sizes of the bran could be beneficial. Thus, millet bran was ground for 2, 4, 8 and 12 minutes using the ball mill (CryoMill, Retsch GmbH, Haan, Germany) at three grinding modes: (I) without cooling, (II) at cryo-temperature, and (III) at cryo-temperature with included 1 minute of intermediate cryo-cooling step after every 2 minutes of grinding, which is usually applied when samples require longer grinding times. The sample was placed in a 50 mL stainless steel jar containing one grinding ball (Ø 25 mm). The oscillation frequency in all three modes was 30 Hz. Particle size distributions of the bran were determined by a laser diffraction particle sizing method (Mastersizer 2000) using the Scirocco 2000 dry dispersion unit (Malvern Instruments, Malvern, UK). Three main effects of the grinding set-up were visible from the results. Firstly, grinding time at all three modes had a significant effect on all particle size parameters: d(0.1), d(0.5), d(0.9), D[3,2], D[4,3], span and specific surface area. Longer grinding times resulted in lower values of the above-listed parameters, e.g. the averaged d(0.5) of the sample (229.57±1.46 µm) dropped to 51.29±1.28 µm after 2 minutes grinding without LN₂, and additionally to 43.00±1.33 µm after 4 minutes of grinding without LN₂. The only exception was the sample ground for 12 minutes without cooling, where an increase in particle diameters occurred (d(0.5)=62.85±2.20 µm), probably due to particles adhering to one another and forming larger particle clusters. Secondly, samples with LN₂ cooling exhibited lower diameters in comparison to non-cooled. For example, after 8 minutes of non-cooled grinding d(0.5)=46.97±1.05 µm was achieved, while the LN₂ cooling enabled collection of particles with average sizes of d(0.5)=18.57±0.18 µm. Thirdly, the application of intermediate cryo-cooling step resulted in similar particle diameters (d(0.5)=15.83±0.36 µm, 12 min of grinding) as cryo-milling without this step (d(0.5)=16.33±2.09 µm, 12 min of grinding). This indicates that intermediate cooling is not necessary for the current application, which consequently reduces the consumption of LN₂. These results point out the potential beneficial effects of millet bran grinding at cryo-temperatures. Further research will show if the lower particle size achieved in comparison to non-cooled grinding could result in increased bioavailability of bioactive compounds, as well as protein digestibility and solubility of dietary fibers of the proso millet bran fraction.

Keywords: ball mill, cryo-milling, particle size distribution, proso millet (Panicum miliaceum) bran

Procedia PDF Downloads 145
75 Technology for Biogas Upgrading with Immobilized Algae Biomass

Authors: Marcin Debowski, Marcin Zielinski, Miroslaw Krzemieniewski, Agata Glowacka-Gil, Paulina Rusanowska, Magdalena Zielinska, Agnieszka Cydzik-Kwiatkowska

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Technologies of biogas upgrading are now perceived as competitive solution combustion and production of electricity and heat. Biomethane production will ensure broader application as energy carrier than biogas. Biomethane can be used as fuel in internal combustion engines or introduced into the natural gas transmission network. Therefore, there is a need to search for innovative, economically and technically justified methods for biogas enrichment. The aim of this paper is to present a technology solution for biogas upgrading with immobilized algae biomass. Reactor for biogas upgrading with immobilized algae biomass can be used for removing CO₂ from the biogas, flue gases and the waste gases especially coming from different industry sectors, e.g. from the food industry from yeast production process, biogas production systems, liquid and gaseous fuels combustion systems, hydrocarbon processing technology. The basis for the technological assumptions of presented technology were laboratory works and analyses that tested technological variants of biogas upgrading. The enrichment of biogas with a methane content of 90-97% pointed to technological assumptions for installation on a technical scale. Reactor for biogas upgrading with algae biomass is characterized by a significantly lower cubature in relation to the currently used solutions which use CO₂ removal processes. The invention, by its structure, assumes achieving a very high concentration of biomass of algae through its immobilization in capsules. This eliminates the phenomenon of lowering the pH value, i.e. acidification of the environment in which algae grow, resulting from the introduction of waste gases at a high CO₂ concentration. The system for introducing light into algae capsules is characterized by a higher degree of its use, due to lower losses resulting from the phenomenon of absorption of light energy by water. The light from the light source is continuously supplied to the formed biomass of algae or cyanobacteria in capsules by the light tubes. The light source may be sunlight or a light generator of a different wavelength of light from 300 nm to 800 nm. A portion of gas containing CO₂, accumulated in the tank and conveyed by the pump is periodically introduced into the housing of the photobioreactor tank. When conveying the gas that contains CO₂, it penetrates the algal biomass in capsules through the outer envelope, displacing, from the algal biomass, gaseous metabolic products which are discharged by the outlet duct for gases. It contributes to eliminating the negative impact of this factor on CO₂ binding processes. As a result of the cyclic dosing of gases containing carbon dioxide, gaseous metabolic products of algae are displaced and removed outside the technological system. Technology for biogas upgrading with immobilized algae biomass is suitable for the small biogas plant. The advantages of this technology are high efficiency as well as useful algae biomass which can be used mainly as animal feed, fertilizers and in the power industry. The construction of the device allows effective removal of carbon dioxide from gases at a high CO₂ concentration.

Keywords: biogas, carbon dioxide, immobilised biomass, microalgae, upgrading

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74 Foslip Loaded and CEA-Affimer Functionalised Silica Nanoparticles for Fluorescent Imaging of Colorectal Cancer Cells

Authors: Yazan S. Khaled, Shazana Shamsuddin, Jim Tiernan, Mike McPherson, Thomas Hughes, Paul Millner, David G. Jayne

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Introduction: There is a need for real-time imaging of colorectal cancer (CRC) to allow tailored surgery to the disease stage. Fluorescence guided laparoscopic imaging of primary colorectal cancer and the draining lymphatics would potentially bring stratified surgery into clinical practice and realign future CRC management to the needs of patients. Fluorescent nanoparticles can offer many advantages in terms of intra-operative imaging and therapy (theranostic) in comparison with traditional soluble reagents. Nanoparticles can be functionalised with diverse reagents and then targeted to the correct tissue using an antibody or Affimer (artificial binding protein). We aimed to develop and test fluorescent silica nanoparticles and targeted against CRC using an anti-carcinoembryonic antigen (CEA) Affimer (Aff). Methods: Anti-CEA and control Myoglobin Affimer binders were subcloned into the expressing vector pET11 followed by transformation into BL21 Star™ (DE3) E.coli. The expression of Affimer binders was induced using 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG). Cells were harvested, lysed and purified using nickle chelating affinity chromatography. The photosensitiser Foslip (soluble analogue of 5,10,15,20-Tetra(m-hydroxyphenyl) chlorin) was incorporated into the core of silica nanoparticles using water-in-oil microemulsion technique. Anti-CEA or control Affs were conjugated to silica nanoparticles surface using sulfosuccinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (sulfo SMCC) chemical linker. Binding of CEA-Aff or control nanoparticles to colorectal cancer cells (LoVo, LS174T and HC116) was quantified in vitro using confocal microscopy. Results: The molecular weights of the obtained band of Affimers were ~12.5KDa while the diameter of functionalised silica nanoparticles was ~80nm. CEA-Affimer targeted nanoparticles demonstrated 9.4, 5.8 and 2.5 fold greater fluorescence than control in, LoVo, LS174T and HCT116 cells respectively (p < 0.002) for the single slice analysis. A similar pattern of successful CEA-targeted fluorescence was observed in the maximum image projection analysis, with CEA-targeted nanoparticles demonstrating 4.1, 2.9 and 2.4 fold greater fluorescence than control particles in LoVo, LS174T, and HCT116 cells respectively (p < 0.0002). There was no significant difference in fluorescence for CEA-Affimer vs. CEA-Antibody targeted nanoparticles. Conclusion: We are the first to demonstrate that Foslip-doped silica nanoparticles conjugated to anti-CEA Affimers via SMCC allowed tumour cell-specific fluorescent targeting in vitro, and had shown sufficient promise to justify testing in an animal model of colorectal cancer. CEA-Affimer appears to be a suitable targeting molecule to replace CEA-Antibody. Targeted silica nanoparticles loaded with Foslip photosensitiser is now being optimised to drive photodynamic killing, via reactive oxygen generation.

Keywords: colorectal cancer, silica nanoparticles, Affimers, antibodies, imaging

Procedia PDF Downloads 239
73 Synthesis and Characterization of Fibrin/Polyethylene Glycol-Based Interpenetrating Polymer Networks for Dermal Tissue Engineering

Authors: O. Gsib, U. Peirera, C. Egles, S. A. Bencherif

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In skin regenerative medicine, one of the critical issues is to produce a three-dimensional scaffold with optimized porosity for dermal fibroblast infiltration and neovascularization, which exhibits high mechanical properties and displays sufficient wound healing characteristics. In this study, we report on the synthesis and characterization of macroporous sequential interpenetrating polymer networks (IPNs) combining skin wound healing properties of fibrin with the excellent physical properties of polyethylene glycol (PEG). Fibrin fibers serve as a provisional biologically active network to promote cell adhesion and proliferation while PEG provides the mechanical stability to maintain the entire 3D construct. After having modified both PEG and Serum Albumin (used for promoting enzymatic degradability) by adding methacrylate residues (PEGDM and SAM, respectively), Fibrin/PEGDM-SAM sequential IPNs were synthesized as follows: Macroporous sponges were first produced from PEGDM-SAM hydrogels by a freeze-drying technique and then rehydrated by adding the fibrin precursors. Environmental Scanning Electron Microscopy (ESEM) and Confocal Laser Scanning Microscopy (CLSM) were used to characterize their microstructure. Human dermal fibroblasts were cultivated during one week in the constructs and different cell culture parameters (viability, morphology, proliferation) were evaluated. Subcutaneous implantations of the scaffolds were conducted on five-week old male nude mice to investigate their biocompatibility in vivo. We successfully synthesized interconnected and macroporous Fibrin/PEGDM-SAM sequential IPNs. The viability of primary dermal fibroblasts was well maintained (above 90%) after 2 days of culture. Cells were able to adhere, spread and proliferate in the scaffolds suggesting the suitable porosity and intrinsic biologic properties of the constructs. The fibrin network adopted a spider web shape that covered partially the pores allowing easier cell infiltration into the macroporous structure. To further characterize the in vitro cell behavior, cell proliferation (EdU incorporation, MTS assay) is being studied. Preliminary histological analysis of animal studies indicated the persistence of hydrogels even after one-month post implantation and confirmed the absence of inflammation response, good biocompatibility and biointegration of our scaffolds within the surrounding tissues. These results suggest that our Fibrin/PEGDM-SAM IPNs could be considered as potential candidates for dermis regenerative medicine. Histological analysis will be completed to further assess scaffold remodeling including de novo extracellular matrix protein synthesis and early stage angiogenesis analysis. Compression measurements will be conducted to investigate the mechanical properties.

Keywords: fibrin, hydrogels for dermal reconstruction, polyethylene glycol, semi-interpenetrating polymer network

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72 Moringa olifera Curate The Toxic Potential of CuO Nanoparticles in Oreochromis mossambicus

Authors: Farhat Jabeen, Muhammad Asad

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The study assessed the curative potential of Moringa olifera seeds against copper oxide nanoparticles induced toxicity in Oreochromis mossambicus. In order to investigate the curative potential of M. olifera seeds, firstly we examine its chemical composition, secondary metabolites, and bioactive compounds including hydroxyl-cinnamic acids, flavanols and hydroxybenzoic acids through standard methods and high performance liquid chromatography. In current study, the potential sub-lethal toxic dose of CuO-NPs (0.12 mg/l) was investigated through pilot experiment and three non-lethal doses (low=32, medium=48 and high=96 mg/l) of M. olifera were selected on the basis of its LC50 value for O. mossambicus. The experimental fish, O. mossambicus (n=100 of approximately 20 g each) were procured from Manawan Fisheries Complex, Lahore, and acclimatized for two weeks in glass aquaria. Experiment was conducted in accordance with the guidelines of Institutional Animal Ethics Committee, Government College University Faisalabad, Pakistan. During acclimatization and experimental period, fish received the commercial fish feed at 2.5% body weight daily. In order to assess the curative effect of M. olifera against CuO NPs induced toxicity, O. mossambicus were randomly divided into five groups and were designated as control (C) without any treatment, positive control (G*) exposed to potential toxic dose of CuO-NPs at 0.12 mg/l, and three treated groups namely G1, G2, and G3 co-treated with 0.12 mg/l of CuO-NPs plus different doses of M. olifera seed extract at 32, 48, and 96 mg/l, respectively for 56 days. Fish were exposed to waterborne CuO NPs and M. olifera seed extract. CuO-NPs treatment was ceased after 28 days but the doses of M. olifera were continued for 56 days. Blood was taken after 28 and 56 days through caudal venipuncture. Liver and intestine were taken for oxidative stress and histological studies after 56 days. In M. olifera seeds, moisture contents, crude protein, lipids, carbohydrates and ash were recorded as 3.8, 37.83, 32.52, 46.12, and 7.75%, respectively on dry weight basis. Total energy was recorded as 627.36 kcal/100g. Qualitative analysis of M. olifera seeds showed the presence of terpenoids, saponins, flavonoids, alkaloids and phenolics, while its quantitative analysis showed the considerable amount of total phenolics, flavonoids, saponins, and alkaloids as 134.75, 170.15, 1.57, and 0.4 µg/mg, respectively. Analysis of bioactive compounds in M. olifera seeds showed the presence of hydroxy-cinnamic acids (6.07 µg/ml), flavanols (71.72 µg/ml), and hydroxyl benzoic acids (97.82 µg/ml). The results showed that M. oliefera seed extract at 48 and 56 mg/l was able to cure against the toxic effects of CuO-NPs. The significant changes were observed in G* and G1 for sero-hepatic enzymes, anti-oxidants and histological profile. The investigations of this study showed that M. olifera is a good curative agent against potential induced toxicity of CuO-NPs in O. mossambicus. The curative effect of M. olifera is attributed to the presence of higher amount of secondary metabolites and bioactive compounds. This study suggested the use of M. olifera to curate different ailments in fish and other organisms.

Keywords: CuO nanoparticles, curative, Moringa olifera, Oreochromis mossambicus

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71 Review of the Nutritional Value of Spirulina as a Potential Replacement of Fishmeal in Aquafeed

Authors: Onada Olawale Ahmed

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As the intensification of aquaculture production increases on global scale, the growing concern of fish farmers around the world is related to cost of fish production, where cost of feeding takes substantial percentage. Fishmeal (FM) is one of the most expensive ingredients, and its high dependence in aqua-feed production translates to high cost of feeding of stocked fish. However, to reach a sustainable aquaculture, new alternative protein sources including cheaper plant or animal origin proteins are needed to be introduced for stable aqua-feed production. Spirulina is a cyanobacterium that has good nutrient profile that could be useful in aquaculture. This review therefore emphasizes on the nutritional value of Spirulina as a potential replacement of FM in aqua-feed. Spirulina is a planktonic photosynthetic filamentous cyanobacterium that forms massive populations in tropical and subtropical bodies of water with high levels of carbonate and bicarbonate. Spirulina grows naturally in nutrient rich alkaline lake with water salinity ( > 30 g/l) and high pH (8.5–11.0). Its artificial production requires luminosity (photo-period 12/12, 4 luxes), temperature (30 °C), inoculum, water stirring device, dissolved solids (10–60 g/litre), pH (8.5– 10.5), good water quality, and macro and micronutrient presence (C, N, P, K, S, Mg, Na, Cl, Ca and Fe, Zn, Cu, Ni, Co, Se). Spirulina has also been reported to grow on agro-industrial waste such as sugar mill waste effluent, poultry industry waste, fertilizer factory waste, and urban waste and organic matter. Chemical composition of Spirulina indicates that it has high nutritional value due to its content of 55-70% protein, 14-19% soluble carbohydrate, high amount of polyunsaturated fatty acids (PUFAs), 1.5–2.0 percent of 5–6 percent total lipid, all the essential minerals are available in spirulina which contributes about 7 percent (average range 2.76–3.00 percent of total weight) under laboratory conditions, β-carotene, B-group vitamin, vitamin E, iron, potassium and chlorophyll are also available in spirulina. Spirulina protein has a balanced composition of amino acids with concentration of methionine, tryptophan and other amino acids almost similar to those of casein, although, this depends upon the culture media used. Positive effects of spirulina on growth, feed utilization and stress and disease resistance of cultured fish have been reported in earlier studies. Spirulina was reported to replace up to 40% of fishmeal protein in tilapia (Oreochromis mossambicus) diet and even higher replacement of fishmeal was possible in common carp (Cyprinus carpio), partial replacement of fish meal with spirulina in diets for parrot fish (Oplegnathus fasciatus) and Tilapia (Orechromis niloticus) has also been conducted. Spirulina have considerable potential for development, especially as a small-scale crop for nutritional enhancement and health improvement of fish. It is important therefore that more research needs to be conducted on its production, inclusion level in aqua-feed and its possible potential use of aquaculture.

Keywords: aquaculture, spirulina, fish nutrition, fish feed

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70 Cereal Bioproducts Conversion to Higher Value Feed by Using Pediococcus Strains Isolated from Spontaneous Fermented Cereal, and Its Influence on Milk Production of Dairy Cattle

Authors: Vita Krungleviciute, Rasa Zelvyte, Ingrida Monkeviciene, Jone Kantautaite, Rolandas Stankevicius, Modestas Ruzauskas, Elena Bartkiene

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The environmental impact of agricultural bioproducts from the processing of food crops is an increasing concern worldwide. Currently, cereal bran has been used as a low-value ingredient for both human consumption and animal feed. The most popular bioprocessing technologies for cereal bran nutritional and technological functionality increasing are enzymatic processing and fermentation, and the most popular starters in fermented feed production are lactic acid bacteria (LAB) including pediococci. However, the ruminant digestive system is unique, there are billions of microorganisms which help the cow to digest and utilize nutrients in the feed. To achieve efficient feed utilization and high milk yield, the microorganisms must have optimal conditions, and the disbalance of this system is highly undesirable. Pediococcus strains Pediococcus acidilactici BaltBio01 and Pediococcus pentosaceus BaltBio02 from spontaneous fermented rye were isolated (by rep – PCR method), identified, and characterized by their growth (by Thermo Bioscreen C automatic turbidometer), acidification rate (2 hours in 2.5 pH), gas production (Durham method), and carbohydrate metabolism (by API 50 CH test ). Antimicrobial activities of isolated pediococcus against variety of pathogenic and opportunistic bacterial strains previously isolated from diseased cattle, and their resistance to antibiotics were evaluated (EFSA-FEEDAP method). The isolated pediococcus strains were cultivated in barley/wheat bran (90 / 10, m / m) substrate, and developed supplements, with high content of valuable pediococcus, were used for Lithuanian black and white dairy cows feeding. In addition, the influence of supplements on milk production and composition was determined. Milk composition was evaluated by the LactoScope FTIR” FT1.0. 2001 (Delta Instruments, Holland). P. acidilactici BaltBio01 and P. pentosaceus BaltBio02 demonstrated versatile carbohydrate metabolism, grown at 30°C and 37°C temperatures, and acidic tolerance. Isolated pediococcus strains showed to be non resistant to antibiotics, and having antimicrobial activity against undesirable microorganisms. By barley/wheat bran utilisation using fermentation with selected pediococcus strains, it is possible to produce safer (reduced Enterobacteriaceae, total aerobic bacteria, yeast and mold count) feed stock with high content of pediococcus. Significantly higher milk yield (after 33 days) by using pediococcus supplements mix for dairy cows feeding could be obtained, while similar effect by using separate strains after 66 days of feeding could be achieved. It can be stated that barley/wheat bran could be used for higher value feed production in order to increase milk production. Therefore, further research is needed to identify what is the main mechanism of the positive action.

Keywords: barley/wheat bran, dairy cattle, fermented feed, milk, pediococcus

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69 Dietary Factors Contributing to Osteoporosis among Postmenopausal Women in Riyadh Armed Forces Hospital

Authors: Rabab Makki

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Bone mineral density and bone metabolism are affected by various factors such as genetic, endocrine, mechanical and nutritional. Our understanding of nutritional influences on bone health is limited because most studies have focused on calcium. This study investigated the dietary factors which are likely t contribute to Osteoporosis in Saudi post-menopausal women, and correlated it with BMD. This is a case controlled study involved 36 postmenopausal Saudi females selected from the Orthopedics and osteoporosis outpatient clinics, and 25 postmenopausal Saudi females as controls from the primary clinic of Military Hospital in Riyadh. The women were diagnosed as osteoporotic based on the BMD measurement at any site (left femur neck, right femur neck, left total hip or right total hip or spine). Both the controls and the Osteoporotics were over 50 years of age and BMI between 31-34 kg/m2 had 2nd degree obesity, and were not free from other problems such as diabetes, hypertension, etc. Subjects (osteoporotics and controls) were interviewed to called data on demographic characterstics, medical history, dietary intake anthropometry (height and weight) bone mineral density. Blood samples were collected from subjects (Osteoporotics and controls). Analysis of serum calcium, vitamin D, phosphate were done at the main laboratory at Military Hospital Riyadh, by the laboratory technician while BMD was determined at the department of Nuclear Medicine by an expert technician and results were interpreted by radiologist.Data on frequency of consumption of animal food (meat, eggs, poultry and fish) and diary foods (milk, yogurt, cheese) of osteoporotic was less than control. In spite of the low intake there was no association with BMD.In general, the vegetables and fruits were consumed less by the osteoporotics than control. The only fruit which had shown a significant positive correlation is banana with right and left hip BMD total probably due to high potassium and minerals content which likely to prevent bone resorption. Mataziz vegetables combination of wheat showed a significant positive correlation with the same site (total right and left hip). Both osteoporotics abd controls were consuming table sugar. (But the sweet intake showed a significant negative correlation with left neck femur BMD, suggesting sucrose increase urinary calcium loss. Both osteoporotic and controls were consuming Arabic coffee. A negative significant correlation between intake of Arabic coffee and BMD of right neck femur of osteoporosis patient was observed. It could be suggested that increased intake of fruits and vegetables, might promote bone density while high intake of coffee and sugars might affect bone density, no significant correlation was observed between BMD at any site and diary product. We can say the major risk factors are inadequate nutrition. Further studies are needed among Saudi population to confirm these results.

Keywords: osteoporosi, Saudia Arabia, Riyadh Armed Forces, postmenopausal women

Procedia PDF Downloads 408
68 Effect of Supplementation with Fresh Citrus Pulp on Growth Performance, Slaughter Traits and Mortality in Guinea Pigs

Authors: Carlos Minguez, Christian F. Sagbay, Erika E. Ordoñez

Abstract:

Guinea pigs (Cavia porcellus) play prominent roles as experimental models for medical research and as pets. However, in developing countries like South America, the Philippines, and sub-Saharan Africa, the meat of guinea pigs is an economic source of animal protein for the poor and malnourished humans because guinea pigs are mainly fed with forage and do not compete directly with human beings for food resources, such as corn or wheat. To achieve efficient production of guinea pigs, it is essential to provide insurance against vitamin C deficiency. The objective of this research was to investigate the effect of the partial replacement of alfalfa with fresh citrus pulp (Citrus sinensis) in a diet of guinea pigs on the growth performance, slaughter traits and mortality during the fattening period (between 20 and 74 days of age). A total of 300 guinea pigs were housed in collective cages of about ten animals (2 x 1 x 0.4 m) and were distributed into two completely randomized groups. Guinea pigs in both groups were fed ad libitum, with a standard commercial pellet diet (10 MJ of digestible energy/kg, 17% crude protein, 11% crude fiber, and 4.5% crude fat). Control group was supplied with fresh alfalfa as forage. In the treatment group, 30% of alfalfa was replaced by fresh citrus pulp. Growth traits, including body weight (BW), average daily gain (ADG), feed intake (FI), and feed conversion ratio (FCR), were measured weekly. On day 74, the animals were slaughtered, and slaughter traits, including live weight at slaughter (LWS), full gastrointestinal tract weight (FGTW), hot carcass weight (with head; HCW), cold carcass weight (with head; CCW), drip loss percentage (DLP) and dressing out carcass yield percentage (DCY), were evaluated. Contrasts between groups were obtained by calculated generalized least squares values. Mortality was evaluated by Fisher's exact test due to low numbers in some cells. In the first week, there were significant differences in the growth traits BW, ADG, FI, and FCR, which were superior in control group. These differences may have been due to the origin of the young guinea pigs, which, before weaning, were all raised without fresh citrus pulp, and they were not familiarized with the new supplement. In the second week, treatment group had significantly increased ADG compared with control group, which may have been the result of a process of compensatory growth. During subsequent weeks, no significant differences were observed between animals raised in the two groups. Neither were any significant differences observed across the total fattening period. No significant differences in slaughter traits or mortality rate were observed between animals from the two groups. In conclusion, although there were no significant differences in growth performance, slaughter traits, or mortality, the use of fresh citrus pulp is recommended. Fresh citrus pulp is a by-product of orange juice industry and it is cheap or free. Forage made with fresh citrus pulp could reduce about of 30 % the quantity of alfalfa in guinea pig for meat and as consequence, reduce the production costs.

Keywords: fresh citrus, growth, Guinea pig, mortality

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67 Bio-Detoxification of Mycotoxins by Lactic Acid Bacteria from Different Food Matrices

Authors: António Inês, Ana Guimarães, José Maria, Vânia Laranjo, Armando Venâncio, Luís Abrunhosa

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Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP-PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 μg/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 μm pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Keywords: bio-detoxification, lactic acid bacteria, mycotoxins, food and feed

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66 COVID-19: Potential Effects of Nutritional Factors on Inflammation Relief

Authors: Maryam Nazari

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COVID-19 is a respiratory disease triggered by the novel coronavirus, SARS-CoV-2, that has reached pandemic status today. Acute inflammation and immune cells infiltration into lung injuries result in multi-organ failure. The presence of other non-communicable diseases (NCDs) with systemic inflammation derived from COVID-19 may exacerbate the patient's situation and increase the risk for adverse effects and mortality. This pandemic is a novel situation and the scientific community at this time is looking for vaccines or drugs to treat the pathology. One of the biggest challenges is focused on reducing inflammation without compromising the correct immune response of the patient. In this regard, addressing the nutritional factors should not be overlooked not only as a matter of avoiding the presence of NCDs with severe infections but also as an adjunctive way to modulate the inflammatory status of the patients. Despite the pivotal role of nutrition in modifying immune response, due to the novelty of the COVID-19 disease, information about the effects of specific dietary agents is limited in this area. From the macronutrients point of view, protein deficiency (quantity or quality) has negative effects on the number of functional immunoglobulins and gut-associated lymphoid tissue (GALT). High biological value proteins or some amino acids like arginine and glutamine are well known for their ability to augment the immune system. Among lipids, fish oil has the ability to inactivate enveloped viruses, suppress pro-inflammatory prostaglandin production and block platelet-activating factors and their receptors. In addition, protectin D1, which is an Omega-3 PUFAs derivation, is a novel antiviral drug. So it seems that these fatty acids can reduce the severity and/or improve recovery of patients with COVID-19. Carbohydrates with lower glycemic index and fibers are associated with lower levels of inflammatory cytokines (CRP, TNF-α, and IL-6). Short-Chain Fatty acids not only exert a direct anti-inflammatory effect but also provide appropriate gut microbial, which is important in gastrointestinal issues related to COVID-19. From the micronutrients point of view, Vitamins A, C, D, E, iron, magnesium, zinc, selenium and copper play a vital role in the maintenance of immune function. Inadequate status in these nutrients may result in decreased resistance against COVID-19 infection. There are specific bioactive compounds in the diet that interact with the ACE2 receptor, which is the gateway for SARS and SARS-CoV-2, and thus controls the viral infection. Regarding this, the potential benefits of probiotics, resveratrol (a polyphenol found in grape), oleoylethanolamide (derived from oleic acid), and natural peroxisome proliferator-activated receptor γ agonists in foodstuffs (like curcumin, pomegranate, hot pepper) are suggested. Yet, it should be pointed out that most of these results have been reported in animal models and further human studies are needed to be verified.

Keywords: Covid-19, inflammation, nutrition, dietary agents

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65 Review of Concepts and Tools Applied to Assess Risks Associated with Food Imports

Authors: A. Falenski, A. Kaesbohrer, M. Filter

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Introduction: Risk assessments can be performed in various ways and in different degrees of complexity. In order to assess risks associated with imported foods additional information needs to be taken into account compared to a risk assessment on regional products. The present review is an overview on currently available best practise approaches and data sources used for food import risk assessments (IRAs). Methods: A literature review has been performed. PubMed was searched for articles about food IRAs published in the years 2004 to 2014 (English and German texts only, search string “(English [la] OR German [la]) (2004:2014 [dp]) import [ti] risk”). Titles and abstracts were screened for import risks in the context of IRAs. The finally selected publications were analysed according to a predefined questionnaire extracting the following information: risk assessment guidelines followed, modelling methods used, data and software applied, existence of an analysis of uncertainty and variability. IRAs cited in these publications were also included in the analysis. Results: The PubMed search resulted in 49 publications, 17 of which contained information about import risks and risk assessments. Within these 19 cross references were identified to be of interest for the present study. These included original articles, reviews and guidelines. At least one of the guidelines of the World Organisation for Animal Health (OIE) and the Codex Alimentarius Commission were referenced in any of the IRAs, either for import of animals or for imports concerning foods, respectively. Interestingly, also a combination of both was used to assess the risk associated with the import of live animals serving as the source of food. Methods ranged from full quantitative IRAs using probabilistic models and dose-response models to qualitative IRA in which decision trees or severity tables were set up using parameter estimations based on expert opinions. Calculations were done using @Risk, R or Excel. Most heterogeneous was the type of data used, ranging from general information on imported goods (food, live animals) to pathogen prevalence in the country of origin. These data were either publicly available in databases or lists (e.g., OIE WAHID and Handystatus II, FAOSTAT, Eurostat, TRACES), accessible on a national level (e.g., herd information) or only open to a small group of people (flight passenger import data at national airport customs office). In the IRAs, an uncertainty analysis has been mentioned in some cases, but calculations have been performed only in a few cases. Conclusion: The current state-of-the-art in the assessment of risks of imported foods is characterized by a great heterogeneity in relation to general methodology and data used. Often information is gathered on a case-by-case basis and reformatted by hand in order to perform the IRA. This analysis therefore illustrates the need for a flexible, modular framework supporting the connection of existing data sources with data analysis and modelling tools. Such an infrastructure could pave the way to IRA workflows applicable ad-hoc, e.g. in case of a crisis situation.

Keywords: import risk assessment, review, tools, food import

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64 Energy Metabolism and Mitochondrial Biogenesis in Muscles of Rats Subjected to Cold Water Immersion

Authors: Bosiacki Mateusz, Anna Lubkowska, Dariusz Chlubek, Irena Baranowska-Bosiacka

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Exposure to cold temperatures can be considered a stressor that can lead to adaptive responses. The present study hypothesized the possibility of a positive effect of cold water exercise on mitochondrial biogenesis and muscle energy metabolism in aging rats. The purpose of this study was to evaluate the effects of cold water exercise on energy status, purine compounds, and mitochondrial biogenesis in the muscles of aging rats as indicators of the effects of cold water exercise and their usefulness in monitoring adaptive changes. The study was conducted on 64 aging rats of both sexes, 15 months old at the time of the experiment. The rats (male and female separately) were randomly assigned to the following study groups: control, sedentary animals; 5°C groups animals - training swimming in cold water at 5°C; 36°C groups - animals training swimming in water at thermal comfort temperature. The study was conducted with the approval of the Local Ethical Committee for Animal Experiments. The animals in the experiment were subjected to swimming training for 9 weeks. During the first week of the study, the duration of the first swimming training was 2 minutes (on the first day), increasing daily by 0.5 minutes up to 4 minutes on the fifth day of the first week. From the second to the eighth week, the swimming training was 4 minutes per day, five days a week. At the end of the study, forty-eight hours after the last swim training, the animals were dissected. In the skeletal muscle tissue of the thighs of the rats, we determined the concentrations of ATP, ADP, AMP, Ado (HPLC), PGC-1a protein expression (Western blot), PGC1A, Mfn1, Mfn2, Opa1, and Drp1 gene expression (qRT PCR). The study showed that swimming in water at a thermally comfortable temperature improved the energy metabolism of the aging rat muscles by increasing the metabolic rate (increase in ATP, ADP, TAN, AEC) and enhancing mitochondrial fusion (increase in mRNA expression of regulatory proteins Mfn1 and Mfn2). Cold water swimming improved muscle energy metabolism in aging rats by increasing the rate of muscle energy metabolism (increase in ATP, ADP, TAN, AEC concentrations) and enhancing mitochondrial biogenesis and dynamics (increase in the mRNA expression of proteins of fusion-regulating factors – Mfn1, Mfn2, and Opa1, and the factor regulating mitochondrial fission – Drp1). The concentration of high-energy compounds and the expression of proteins regulating mitochondrial dynamics in the muscle may be a useful indicator in monitoring adaptive changes occurring in aging muscles under the influence of exercise in cold water. It represents a short-term adaptation to changing environmental conditions and has a beneficial effect on maintaining the bioenergetic capacity of muscles in the long term. Conclusion: exercise in cold water can exert positive effects on energy metabolism, biogenesis and dynamics of mitochondria in aging rat muscles. Enhancement of mitochondrial dynamics under cold water exercise conditions can improve mitochondrial function and optimize the bioenergetic capacity of mitochondria in aging rat muscles.

Keywords: cold water immersion, adaptive responses, muscle energy metabolism, aging

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63 Glucose Measurement in Response to Environmental and Physiological Challenges: Towards a Non-Invasive Approach to Study Stress in Fishes

Authors: Tomas Makaras, Julija Razumienė, Vidutė Gurevičienė, Gintarė Sauliutė, Milda Stankevičiūtė

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Stress responses represent animal’s natural reactions to various challenging conditions and could be used as a welfare indicator. Regardless of the wide use of glucose measurements in stress evaluation, there are some inconsistencies in its acceptance as a stress marker, especially when it comes to comparison with non-invasive cortisol measurements in the fish challenging stress. To meet the challenge and to test the reliability and applicability of glucose measurement in practice, in this study, different environmental/anthropogenic exposure scenarios were simulated to provoke chemical-induced stress in fish (14-days exposure to landfill leachate) followed by a 14-days stress recovery period and under the cumulative effect of leachate fish subsequently exposed to pathogenic oomycetes (Saprolegnia parasitica) to represent a possible infection in fish. It is endemic to all freshwater habitats worldwide and is partly responsible for the decline of natural freshwater fish populations. Brown trout (Salmo trutta fario) and sea trout (Salmo trutta trutta) juveniles were chosen because of a large amount of literature on physiological stress responses in these species was known. Glucose content in fish by applying invasive and non-invasive glucose measurement procedures in different test mediums such as fish blood, gill tissues and fish-holding water were analysed. The results indicated that the quantity of glucose released in the holding water of stressed fish increased considerably (approx. 3.5- to 8-fold) and remained substantially higher (approx. 2- to 4-fold) throughout the stress recovery period than the control level suggesting that fish did not recover from chemical-induced stress. The circulating levels of glucose in blood and gills decreased over time in fish exposed to different stressors. However, the gill glucose level in fish showed a decrease similar to the control levels measured at the same time points, which was found to be insignificant. The data analysis showed that concentrations of β-D glucose measured in gills of fish treated with S. parasitica differed significantly from the control recovery, but did not differ from the leachate recovery group showing that S. parasitica presence in water had no additive effects. In contrast, a positive correlation between blood and gills glucose were determined. Parallel trends in blood and water glucose changes suggest that water glucose measurement has much potency in predicting stress. This study demonstrated that measuring β-D-glucose in fish-holding water is not stressful as it involves no handling and manipulation of an organism and has critical technical advantages concerning current (invasive) methods, mainly using blood samples or specific tissues. The quantification of glucose could be essential for studies examining the stress physiology/aquaculture studies interested in the assessment or long-term monitoring of fish health.

Keywords: brown trout, landfill leachate, sea trout, pathogenic oomycetes, β-D-glucose

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62 Mixed Monolayer and PEG Linker Approaches to Creating Multifunctional Gold Nanoparticles

Authors: D. Dixon, J. Nicol, J. A. Coulter, E. Harrison

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The ease with which they can be functionalized, combined with their excellent biocompatibility, make gold nanoparticles (AuNPs) ideal candidates for various applications in nanomedicine. Indeed several promising treatments are currently undergoing human clinical trials (CYT-6091 and Auroshell). A successful nanoparticle treatment must first evade the immune system, then accumulate within the target tissue, before enter the diseased cells and delivering the payload. In order to create a clinically relevant drug delivery system, contrast agent or radiosensitizer, it is generally necessary to functionalize the AuNP surface with multiple groups; e.g. Polyethylene Glycol (PEG) for enhanced stability, targeting groups such as antibodies, peptides for enhanced internalization, and therapeutic agents. Creating and characterizing the biological response of such complex systems remains a challenge. The two commonly used methods to attach multiple groups to the surface of AuNPs are the creation of a mixed monolayer, or by binding groups to the AuNP surface using a bi-functional PEG linker. While some excellent in-vitro and animal results have been reported for both approaches further work is necessary to directly compare the two methods. In this study AuNPs capped with both PEG and a Receptor Mediated Endocytosis (RME) peptide were prepared using both mixed monolayer and PEG linker approaches. The PEG linker used was SH-PEG-SGA which has a thiol at one end for AuNP attachment, and an NHS ester at the other to bind to the peptide. The work builds upon previous studies carried out at the University of Ulster which have investigated AuNP synthesis, the influence of PEG on stability in a range of media and investigated intracellular payload release. 18-19nm citrate capped AuNPs were prepared using the Turkevich method via the sodium citrate reduction of boiling 0.01wt% Chloroauric acid. To produce PEG capped AuNPs, the required amount of PEG-SH (5000Mw) or SH-PEG-SGA (3000Mw Jenkem Technologies) was added, and the solution stirred overnight at room temperature. The RME (sequence: CKKKKKKSEDEYPYVPN, Biomatik) co-functionalised samples were prepared by adding the required amount of peptide to the PEG capped samples and stirring overnight. The appropriate amounts of PEG-SH and RME peptide were added to the AuNP to produce a mixed monolayer consisting of approximately 50% PEG and 50% RME. The PEG linker samples were first fully capped with bi-functional PEG before being capped with RME peptide. An increase in diameter from 18-19mm for the ‘as synthesized’ AuNPs to 40-42nm after PEG capping was observed via DLS. The presence of PEG and RME peptide on both the mixed monolayer and PEG linker co-functionalized samples was confirmed by both FTIR and TGA. Bi-functional PEG linkers allow the entire AuNP surface to be capped with PEG, enabling in-vitro stability to be achieved using a lower molecular weight PEG. The approach also allows the entire outer surface to be coated with peptide or other biologically active groups, whilst also offering the promise of enhanced biological availability. The effect of mixed monolayer versus PEG linker attachment on both stability and non-specific protein corona interactions was also studied.

Keywords: nanomedicine, gold nanoparticles, PEG, biocompatibility

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61 Human Facial Emotion: A Comparative and Evolutionary Perspective Using a Canine Model

Authors: Catia Correia Caeiro, Kun Guo, Daniel Mills

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Despite its growing interest, emotions are still an understudied cognitive process and their origins are currently the focus of much debate among the scientific community. The use of facial expressions as traditional hallmarks of discrete and holistic emotions created a circular reasoning due to a priori assumptions of meaning and its associated appearance-biases. Ekman and colleagues solved this problem and laid the foundations for the quantitative and systematic study of facial expressions in humans by developing an anatomically-based system (independent from meaning) to measure facial behaviour, the Facial Action Coding System (FACS). One way of investigating emotion cognition processes is by applying comparative psychology methodologies and looking at either closely-related species (e.g. chimpanzees) or phylogenetically distant species sharing similar present adaptation problems (analogy). In this study, the domestic dog was used as a comparative animal model to look at facial expressions in social interactions in parallel with human facial expressions. The orofacial musculature seems to be relatively well conserved across mammal species and the same holds true for the domestic dog. Furthermore, the dog is unique in having shared the same social environment as humans for more than 10,000 years, facing similar challenges and acquiring a unique set of socio-cognitive skills in the process. In this study, the spontaneous facial movements of humans and dogs were compared when interacting with hetero- and conspecifics as well as in solitary contexts. In total, 200 participants were examined with FACS and DogFACS (The Dog Facial Action Coding System): coding tools across four different emotionally-driven contexts: a) Happiness (play and reunion), b) anticipation (of positive reward), c) fear (object or situation triggered), and d) frustration (negation of a resource). A neutral control was added for both species. All four contexts are commonly encountered by humans and dogs, are comparable between species and seem to give rise to emotions from homologous brain systems. The videos used in the study were extracted from public databases (e.g. Youtube) or published scientific databases (e.g. AM-FED). The results obtained allowed us to delineate clear similarities and differences on the flexibility of the facial musculature in the two species. More importantly, they shed light on what common facial movements are a product of the emotion linked contexts (the ones appearing in both species) and which are characteristic of the species, revealing an important clue for the debate on the origin of emotions. Additionally, we were able to examine movements that might have emerged for interspecific communication. Finally, our results are discussed from an evolutionary perspective adding to the recent line of work that supports an ancient shared origin of emotions in a mammal ancestor and defining emotions as mechanisms with a clear adaptive purpose essential on numerous situations, ranging from maintenance of social bonds to fitness and survival modulators.

Keywords: comparative and evolutionary psychology, emotion, facial expressions, FACS

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60 Enhanced Functional Production of a Crucial Biomolecule Human Serum Albumin in Escherichia coli

Authors: Ashima Sharma

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Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

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