Search results for: T. Ferreira

Authors: Rafael Felipe Guatura da Silva, José Luis Gomes da Silva, Luiz Antonio, Ferreira Perrone de Brito

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Noise can cause serious damage to human health, such as hearing loss, stress, irritability, fatigue, and others. Aviation is a place where your entire process should be work out with the utmost attention and commitment of human resources, thus the need to study the effects of noise in this sector, as aeronautical noise levels are high. This study aimed to evaluate the impact of noise pollution on the performance of professionals regarding the fatigue generated by aeronautical noise and time to noise exposure. The methodology used consists of measurements of sound pressure levels at 42 points of the aerodrome. The selected points are located inside the hangars and outside the airfield hangars. All points chosen are close to the professionals' work areas, seeking to identify the sound pressure levels to which they submitted. The other part of the research used the principle on the application of a self-report questionnaire to a sample of 207 people working inside the aerodrome. The 207 professionals surveyed consist of aircraft mechanics, pilots, maintenance managers, and administrative professionals. The questionnaire was intended to evaluate the knowledge that professionals have about health risks caused by sound exposure as well as to identify diseases that professionals have, and that may be associated with exposure to high levels of sound pressure. Preliminary results identify points with sound pressure levels of up to 91.7 dB, thus highlighting the need for the use of personal protective equipment that reduces noise exposure. It was also identified a large number of professionals who are bothered by the sound exposure and approximately 25% of professionals interviewed reported having a hearing disorder.

Keywords: aeronautical noise, fatigue, noise and health, noise management

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Authors: Renata Rank Miranda, Arandi Ginane Bezerra, Ciro Alberto Oliveira Ribeiro, Marco AntôNio Ferreira Randi, Carmen Lúcia Voigt, Lilian Skytte, Kaare Lund Rasmussen, Francisco Filipak Neto, Frank Kjeldsen

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Synergetic and antagonistic effects of drugs are well-known concerns in pharmacological assessments of dose and toxicity. Similar approach should be used in assessing cellular uptake and cytotoxicity of nanoparticles. Since nanoparticles are released into the aquatic environment they may interact with existing xenobiotics. Here we used biochemical assays and quantitative proteomics to assess the cytotoxicity of silver nanoparticles (AgNP) when human hepatoma HepG2 cells were co-exposed to 2 nm AgNP together with either Cd2+ or Hg2+ ions. Time-course experiments (2h, 4h, and 24h) were conducted to assess the first response to the exposure studies. The general trend was that a synergetic toxicological response was observed in cells exposed to both AgNP and Cd2+ or Hg2+, with AgNP and Cd2+ being more toxic. This was observed by a significant increase in the ROS and superoxide level of >35% in the case of AgNP+Cd2+ compared to the sum of responses of AgNP and Cd2+, individually. Metabolic activity and viability also dropped more for AgNP+Cd2+ (>10%) than for AgNP and Cd2+ combined. We used inductively coupled plasma mass spectrometry to investigate if AgNP facilitates larger influx of toxic metal ions into HepG2 cells. Only Hg2+ ions was found to be more efficiently engulfed as the concentration of Hg2+ was found 2.8 times larger compared to exposure experiments with only Hg2+. This effect was not observed for Cd2+. We now continue with deep proteomics studies to obtain wider details on the mechanism of the toxicity related to AgNP, Cd2+, and AgNP+Cd2+, respectively.

Keywords: nanotoxicology, silver nanoparticles, proteomics, human cell line

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Authors: Tahir Khaleeq, Patrick Lancaster, Keji Fakoya, Pedro Ferreira, Usman Ahmed

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Introduction: Virtual fracture clinics (VFC) have been shown to be a safe and cost-effective way of managing outpatient referrals to the orthopaedic department. During the coronavirus pandemic there has been a push to reduce unnecessary patient contact whilst maintaining patient safety. Materials and Methods: A protocol was developed by the clinical team in collaboration with Advanced Physiotherapy Practitioners (APP) on how to manage common musculoskeletal presentations to A&E prior to COVID as part of routine service development. Patients broadly triaged into 4 categories; discharge with advice, referral to VFC, referral to face to face clinic or discussion with on call team. The first 9 months of data were analysed to assess types of injury seen and outcomes. Results: In total 2489 patients were referred to VFC from internal and external sources. 734 patients were discharged without follow-up and 182 patients were discharged for physiotherapy review. Only 3 patients required admission. Regarding follow-ups, 431 patients had a virtual follow-up while 1036 of patients required further face to face follow up. 87 patients were triaged into subspecialty clinics. 37 patients were felt to have been referred inappropriately. Discussion: BOA guidelines suggest all patients need to be reviewed within 72 hours of their orthopaedic injury. Implementation of a VFC allows this target to be achieved and at the same time reduce patient contact. Almost half the patients were discharged following VFC review, the remaining patients were appropriately followed up. This is especially relevant in the current pandemic where reducing unnecessary trips to hospital will benefit the patient as well as make the most of the resources available.

Keywords: virtual fracture clinic, lockdown, trauma and orthopaedics, Covid- 19

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Authors: Luísa P. Cruz-Lopes, Hugo Costa e Silva, Idalina Domingos, José Ferreira, Luís Teixeira de Lemos, Bruno Esteves

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The liquefaction process of cork based tree barks has led to an increase of interest due to its potential innovation in the lumber and wood industries. In this particular study the bark of Quercus cerris (Turkish oak) is used due to its appreciable amount of cork tissue, although of inferior quality when compared to the cork provided by other Quercus trees. This study aims to optimize alkaline catalysis liquefaction conditions, regarding several parameters. To better comprehend the possible chemical characteristics of the bark of Quercus cerris, a complete chemical analysis was performed. The liquefaction process was performed in a double-jacket reactor heated with oil, using glycerol and a mixture of glycerol/ethylene glycol as solvents, potassium hydroxide as a catalyst, and varying the temperature, liquefaction time and granulometry. Due to low liquefaction efficiency resulting from the first experimental procedures a study was made regarding different washing techniques after the filtration process using methanol and methanol/water. The chemical analysis stated that the bark of Quercus cerris is mostly composed by suberin (ca. 30%) and lignin (ca. 24%) as well as insolvent hemicelluloses in hot water (ca. 23%). On the liquefaction stage, the results that led to higher yields were: using a mixture of methanol/ethylene glycol as reagents and a time and temperature of 120 minutes and 200 ºC, respectively. It is concluded that using a granulometry of <80 mesh leads to better results, even if this parameter barely influences the liquefaction efficiency. Regarding the filtration stage, washing the residue with methanol and then distilled water leads to a considerable increase on final liquefaction percentages, which proves that this procedure is effective at liquefying suberin content and lignocellulose fraction.

Keywords: liquefaction, Quercus cerris, polyalcohol liquefaction, temperature

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Authors: Pedro Ferreira, Alexandre Frugoli, Pedro Frugoli, Lucio Leonardo, Thais Cavalheri

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The teaching of fluid mechanics in engineering courses is, in general, a source of great difficulties for learning. The possibility of the use of experiments with didactic wind tunnels can facilitate the education of future professionals. The objective of this proposal is the development of a low-cost aerodynamic balance to be used in a didactic wind tunnel. The set is comprised of an Arduino microcontroller, programmed by an open source software, linked to load cells built by students from another project. The didactic wind tunnel is 5,0m long and the test area is 90,0 cm x 90,0 cm x 150,0 cm. The Weq® electric motor, model W-22 of 9,2 HP, moves a fan with nine blades, each blade 32,0 cm long. The Weq® frequency inverter, model WEGCFW 08 (Vector Inverter) is responsible for wind speed control and also for the motor inversion of the rotational direction. A flat-convex profile prototype of airfoil was tested by measuring the drag and lift forces for certain attack angles; the air flux conditions remained constant, monitored by a Pitot tube connected to a EXTECH® Instruments digital pressure differential manometer Model HD755. The results indicate a good agreement with the theory. The choice of all of the components of this proposal resulted in a low-cost product providing a high level of specific knowledge of mechanics of fluids, which may be a good alternative to teaching in countries with scarce educational resources. The system also allows the expansion to measure other parameters like fluid velocity, temperature, pressure as well as the possibility of automation of other functions.

Keywords: aerodynamic balance, wind tunnel, strain gauge, load cell, Arduino, low-cost education

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Authors: Nunila Ferreira de Oliveira, Silvana Martins Mishima

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Diabetes mellitus type 2 (DM2) prevalence, is increasing on the world, in Brazil is considered a public health problem. Treatment focuses on glycemic control depending primarily of lifestyle changes - not drug treatment (NDT), may involve drug therapy (DT) and requires continuous health monitoring. In Brazil this monitoring is performed by the Unified Health System (SUS) through Primary Health Care (PHC), which stimulate people with DM2 empowerment for care management. SUS was approved in 1988 and the PHC operationalization was strengthened with the creation of the Family Health Strategy (FHS) in 1994. Our aim was to analyze the people with DM2 participation in front of the care management health monitoring in the FHS. Qualitative research was carried out through non-participant observation of attendance of 25 people with DM2 in the FHS and interviewed at home. Ethical guidelines were followed. It was found that people with DM2 only follow professionals’ recommendations that make sense according to their own conceptions of health/disease; most of them emphasize the importance of (DT) with little emphasis on the NDT, was found great difficulty in the NDT and lack of knowledge about the disease and care. As regards monitoring the FHS, were observed therapeutic practices based on the bio medical model, although the APS search for another care perspective; NDT is not systematically accompanied by the health team and takes place a few educational activities on the DM2 in the FHS, with low user adoption. The work of the FHS is done by multidisciplinary teams, but we see the need for greater participation of nurses in clinical-care follow-up of this population and may also act in adapting to the NDT. Finally we emphasize the need for professional practices that consider the difficulties to care management by people with DM2, especially because of the NDT. It is noticed that the measures recommended by the FHS professionals are not always developed by people with DM2. We must seek the empowerment of people with DM2 to manage the form of care associated with the FHS team, seeking to reduce the incidence of complications and higher quality of life.

Keywords: diabetes mellitus, primary health care, nursing, management of care

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Authors: Fabiane Alves Farias Guimarães, Rodrigo Otávio Moretti-Pires, Ana Lúcia Schaefer Ferreira de Mello

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Introduction: Considering the last changes in Brazilian Health and Higher Educational Systems, the production of scientific knowledge regarding dental education and training has been increasing. The National Curriculum Guidelines for undergraduate courses in Dentistry established in 2002 the principles and procedures to perform a more generalist dental professional profile. Objectives: To perform a systematic review of the Brazilian scientific literature about dental education and training. Methods: The systematic review was conducted considering the Lilacs - Latin American Literature in Health Sciences and SciELO - Scientific Electronic Library Online data bases, using the combination of key words dentistry, education, teaching or training. It was select original research articles, published between 2010 and 2013, in Portuguese. Results: Based on the selection criteria, it was found 23 articles. In order to organize the outcomes, the analysis was separated in three themes: Ethical aspects of education (3 articles), integrating dental service with training (10 articles) and Dental education and the Brazilian curriculum guidelines (10 articles). Most of the studies were published between 2011 and 2012 (35% each) and were held in public universities. The studied populations included dental students, teachers, universities directors, health managers and dentists. The qualitative methodological approach was predominant. Conclusion: It was possible to identify a transience time in Brazilian undergraduate courses in Dentistry after curricular changes. The produced literature shows some advances, as the incorporation of ethical values on dental education and the inclusion of new practices environments for students by integrating education and training in diversified dental services scenarios.

Keywords: Teaching, Dental Students, Human resources in dentistry

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Authors: Carla Maria Costa Ferreira, Maria Helena Freitas Melao Barros

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The rules implemented in Europe regarding structural design are termed Structural Eurocodes and deal with the several materials available for construction. Particularly regarding the very used in Europe concrete with steel reinforcement, it is named the Eurocode 2 – Design of Concrete Structures, usually known as EC2. The need of tables and abacuses to help in the design of reinforced concrete was due to the fact that the evolution and the study of new procedures and higher strength concrete showed that the previous tables needed to be improved. Reinforced concrete structures have particular aspects in the design that come from the nonlinear behavior of the concrete and steel and, in the case of concrete, also by the very low tensile strength. The design of reinforced concrete structures is made in terms of evaluating the ultimate strength and how it behaves under service conditions. As a matter of fact, the use of higher-strength concrete and steel classes showed that these serviceability design that was important for prestressed structures may be relevant in reinforced concrete structures. For these aspects, there are tables and design charts used for the ultimate limit design of reinforced concrete sections under bending moments and axial forces, and also auxiliary design diagrams able to evaluate the stress of the steel and the concrete at a section and the ductility for service limit states verification. For practical use, here are presented tables and design charts for the ultimate limit design of reinforced concrete sections and also auxiliary interaction diagrams for verification of the serviceability conditions. These kinds of aid for design were only available to engineers before the development of computers and, nowadays, yet an important tool in the universities for the students' use. Usually, in the reinforced concrete design, it is needed to obtain the area of the steel longitudinal reinforcement to be placed in the structure. The quantity and the position of the steel area may have different solutions and these tables and abacuses permit to obtain many possibilities in order to optimize the solution in economic or ductility terms.

Keywords: design examples, Eurocode 2, reinforced concrete, section design

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Authors: Thais Cavalheri Santos, Pedro Jose Gabriel Ferreira, Alexandre Daliberto Frugoli, Lucio Leonardo, Pedro Americo Frugoli

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This paper presents a low cost education proposal to be used in engineering courses. The engineering education in universities of a developing country that is in need of an increasing number of engineers carried out with quality and affordably, pose a difficult problem to solve. In Brazil, the political and economic scenario requires academic managers able to reduce costs without compromising the quality of education. Within this context, the elaboration of a physics principles teaching method with the construction of an electronic balance is proposed. First, a method to develop and construct a load cell through which the students can understand the physical principle of strain gauges and bridge circuit will be proposed. The load cell structure was made with aluminum 6351T6, in dimensions of 80 mm x 13 mm x 13 mm and for its instrumentation, a complete Wheatstone Bridge was assembled with strain gauges of 350 ohms. Additionally, the process involves the use of a software tool to document the prototypes (design circuits), the conditioning of the signal, a microcontroller, C language programming as well as the development of the prototype. The project also intends to use an open-source I/O board (Arduino Microcontroller). To design the circuit, the Fritizing software will be used and, to program the controller, an open-source software named IDE®. A load cell was chosen because strain gauges have accuracy and their use has several applications in the industry. A prototype was developed for this study, and it confirmed the affordability of this educational idea. Furthermore, the goal of this proposal is to motivate the students to understand the several possible applications in high technology of the use of load cells and microcontroller.

Keywords: Arduino, load cell, low-cost education, strain gauge

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Authors: J. F. Pollo Gaspary, F. Peron Gaspary, E. M. Simão, R. Concatto Beltrame, G. Orengo de Oliveira, M. S. Ristow Ferreira, F. Sartori Thies, I. F. Minello, F. dos Santos de Oliveira

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Background: The immune system has evolved several mechanisms to protect the host against cancer, and it has now been suggested that the expansion of its functions may prevent tumor growth and control the symptoms of cancer patients. Two techniques, ozone therapy and pulsed electromagnetic fields (PEMF), are independently associated with an increase in the immune system functions and they maybe help palliative care of patients in these conditions. Case Report: A patient with rectal adenocarcinoma with metastases decides to interrupt the clinical chemotherapy protocol due to refractoriness and side effects. As a palliative care alternative treatment it is suggested to the patient the use of ozone therapy associated with PEMF techniques. Results: The patient reports an improvement in well-being, in autonomy and in pain control. Imaging tests confirm a pause in tumor growth despite more than 60 days without using classic treatment. These results associated with palliative care alternative treatment stimulate the return to the chemotherapy protocol. Discussion: This case illustrates that these two techniques can contribute to the control of tumor growth and refractory symptoms, such as pain, probably by enhancing the immune system. Conclusions: The potential use of the combination of these two therapies, ozone therapy and PEMF therapy, can contribute to palliation of cancer patients, alone or in combination with pharmacological therapies. The conduct of future investigations on this paradigm can elucidate how much these techniques contribute to the survival and well-being of these patients.

Keywords: cancer, complementary and alternative medicine , ozone therapy, palliative care, PEMF therapy

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Authors: Maria L. G. Lourenco, Viviane Y. Hibaru, Keylla H. N. P. Pereira, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Neonatal conjunctivitis, or ophthalmia, is an infection of the conjunctiva or cornea before opening the eyelids. It is believed that immunodeficiency contributes to the development of the condition. This study aims at reporting a case of ophthalmia neonatorum in a dog, in addition to its diagnosis and treatment. A litter of five pug neonates was admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, with complaints of ocular secretion. The neonates were five days old. The clinical examination revealed that three newborns presented swelling in the ocular region and a purulent secretion in the medial corner of the eye that was exerting pressure on the ocular globes, which are compatible with the description of this disease. The diagnosis was made based on the clinical signs and bacterial culture of the secretion, which revealed the presence of bacteria belonging to the genus Staphylococcus sp. The laboratory assays did not reveal any alterations. The treatment was instituted gently, opening the eyelids early and cleaning the purulent ocular secretion with saline solution. An ophthalmic ointment with retinol, amino acids, methionine, and chloramphenicol (Epitezan®) was prescribed four times a day for seven days. Blood plasma (2 mL/100 g) was administered subcutaneously because bacterial infections in neonates may represent a failure in the transference of passive immunity. A more thorough cleaning of the environment was also recommended. Neonatal conjunctivitis has a simple diagnosis and treatment. If not treated early, it can evolve to adherence of the eyelids to the cornea, ulceration, and perforation of the cornea. Therefore, the prognosis is favorable as long as the condition is diagnosed early, and the treatment is instituted quickly.

Keywords: ophthalmia neonatorum, neonatal infection, puppy, newborn

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Authors: Renan De Caldas Honorato, Ana Augusta Ferreira De Freitas

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Homosexuality is still criminalized in a large number of countries. In some of them, being gay or lesbian can even be punished by death. Added to this context, the experiences of social discrimination faced by the LGBT population, including homophobia, machismo and misogyny, cause numerous restrictions throughout their lives. The possibility of confronting these challenges in moments that should be pleasant, such as on a trip or on vacation, is unpleasant, to say the least. In the current scenario of intensifying the use of Social network sites (SNSs) to search for information, including in the tourist area, this work aims to analyze the sharing of tourist experiences with situations of confrontation and perceptions of homophobia, machismo and misogyny, and restrictions suffered in tourist destinations. The fieldwork is a community of LGBT backpackers based on Facebook. Netnography was the core method adopted. A qualitative approach was conducted and 463 publications posted from January to December 2020 were assessed through the computer-mediated discourse analysis (CMDA). The results suggest that these publications exist to identify the potential exposure to these offensive behaviors while traveling. Individuals affirm that the laws, positive or not, in relation to the LGBT public are not the only factors for a place to be defined as safe or not for gay travelers. The social situation of a country and its laws are quite different and this is the main target of these publications. The perception of others about the chosen destination is more important than knowing your rights and the legal status of each country and it also lessens uncertainty, even when they are never totally confident when choosing a travel destination. In certain circumstances, sexual orientation also needs to be protected from the judgment of hosts and residents. The systemic treatment of homophobic behavior and the construction of a more inclusive society are urgent.

Keywords: homophobia, hospitality, machismo, misogyny

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Authors: Jesus D. Cortes Gil, Fernanda Ewerling, Leonardo Ferreira, Aluisio J. D. Barros

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Background: The sustainable development goals call for inclusive, equitable, and quality learning opportunities for all. This is especially important for children, to ensure they all develop to their full potential. We studied the prevalence and inequalities of suspected delay in child development in 63 low- and middle-income countries. Methods and Findings: We used the early child development module from national health surveys, which covers four developmental domains (physical, social-emotional, learning, literacy-numeracy) and provides a combined indicator (early child development index, ECDI) of whether children are on track. We calculated the age-adjusted prevalence of suspected delay at the country level and stratifying by wealth, urban/rural residence, sex of the child, and maternal education. We also calculated measures of absolute and relative inequality. We studied 330.613 children from 63 countries. The prevalence of suspected delay for the ECDI ranged from 3% in Barbados to 67% in Chad. For all countries together, 25% of the children were suspected of developmental delay. At regional level, the prevalence of delay ranged from 10% in Europe and Central Asia to 42% in West and Central Africa. The literacy-numeracy domain was by far the most challenging, with the highest proportions of delay. We observed very large inequalities, and most markedly for the literacy-numeracy domain. Conclusions: To date, our study presents the most comprehensive analysis of child development using an instrument especially developed for national health surveys. With a quarter of the children globally suspected of developmental delay, we face an immense challenge. The multifactorial aspect of early child development and the large gaps we found only add to the challenge of not leaving these children behind.

Keywords: child development, inequalities, global health, equity

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Authors: M. C. Pardo-Ferreira, J. C. Rubio-Romero, M. Martínez-Rojas

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Resilience Engineering is a new paradigm of safety management that proposes to change the way of managing the safety to focus on the things that go well instead of the things that go wrong. Many complex and high-risk sectors such as air traffic control, health care, nuclear power plants, railways or emergencies, have applied this new vision of safety and have obtained very positive results. In the construction sector, safety management continues to be a problem as indicated by the statistics of occupational injuries worldwide. Therefore, it is important to improve safety management in this sector. For this reason, it is proposed to apply Resilience Engineering to the construction sector. The Construction Phase Health and Safety Plan emerges as a key element for the planning of safety management. One of the key tools of Resilience Engineering is the Resilience Assessment Grid that allows measuring the four essential abilities (respond, monitor, learn and anticipate) for resilient performance. The purpose of this paper is to develop a questionnaire based on the Resilience Assessment Grid, specifically on the ability to learn, to assess whether a Construction Phase Health and Safety Plans helps companies in a construction site to implement this ability. The research process was divided into four stages: (i) initial design of a questionnaire, (ii) validation of the content of the questionnaire, (iii) redesign of the questionnaire and (iii) application of the Delphi method. The questionnaire obtained could be used as a tool to help construction companies to evolve from Safety-I to Safety-II. In this way, companies could begin to develop the ability to learn, which will serve as a basis for the development of the other abilities necessary for resilient performance. The following steps in this research are intended to develop other questions that allow evaluating the rest of abilities for resilient performance such as monitoring, learning and anticipating.

Keywords: resilience engineering, construction sector, resilience assessment grid, construction phase health and safety plan

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Authors: Maria João Ferreira, Natalia Sierra-Garcia, Javier Cremades, Carla António, Ana M. Rodrigues, Helena Silva, Ângela Cunha

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Salicornia ramosissima, a halophyte that grows naturally in coastal areas of the northern hemisphere, is often considered the most promising halophyte candidate for extensive crop cultivation and saline agriculture practices. The expanding interest in this plant surpasses its use as gourmet food and includes their potential application as a source of bioactive compounds for the pharmaceutical industry. Despite growing well in saline soils, sustainable and ecologically friendly techniques to enhance crop production and the nutritional value of this plant are still needed. The root microbiome of S. ramosissima proved to be a source of taxonomically diverse plant growth-promoting bacteria (PGPB). Halotolerant strains of Bacillus, Salinicola, Pseudomonas, and Brevibacterium, among other genera, exhibit a broad spectrum of plant-growth promotion traits [e.g., 3-indole acetic acid (IAA), 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, siderophores, phosphate solubilization, Nitrogen fixation] and express a wide range of extracellular enzyme activities. In this work, three plant growth-promoting bacteria strains (Brevibacterium casei EB3, Pseudomonas oryzihabitans RL18, and Bacillus aryabhattai SP20) isolated from the rhizosphere and the endosphere of S. ramosissima roots from different saltmarshes along the Portuguese coast were inoculated in S. ramosissima seeds. Plants germinated from inoculated seeds were grown for three months in pots filled with a mixture of perlite and estuarine sediment (1:1) in greenhouse conditions and later transferred to a growth chamber, where they were maintained two months with controlled photoperiod, temperature, and humidity. Pots were placed on trays containing the irrigation solution (Hoagland’s solution 20% added with 10‰ marine salt). Before reaching the flowering stage, plants were collected, and the fresh and dry weight of aerial parts was determined. Non-inoculated seeds were used as a negative control. Selected dried stems from the most promising treatments were later analyzed by GC-TOF-MS for primary metabolite composition. The efficiency of inoculation and persistence of the inoculum was assessed by Next Generation Sequencing. Inoculations with single strain EB3 and co-inoculations with EB3+RL18 and EB3+RL18+SP20 (All treatment) resulted in significantly higher biomass production (fresh and dry weight) compared to non-inoculated plants. Considering fresh weight alone, inoculation with isolates SP20 and RL18 also caused a significant positive effect. Combined inoculation with the consortia SP20+EB3 or SP20+RL18 did not significantly improve biomass production. The analysis of the profile of primary metabolites will provide clues on the mechanisms by which the growth-enhancement effect of the inoculants operates in the plants. These results sustain promising prospects for the use of rhizospheric and endophytic PGPB as biofertilizers, reducing environmental impacts and operational costs of agrochemicals and contributing to the sustainability and cost-effectiveness of saline agriculture. Acknowledgments: This work was supported by project Rhizomis PTDC/BIA-MIC/29736/2017 financed by Fundação para a Ciência e Tecnologia (FCT) through the Regional Operational Program of the Center (02/SAICT/2017) with FEDER funds (European Regional Development Fund, FNR, and OE) and by FCT through CESAM (UIDP/50017/2020 + UIDB/50017/2020), LAQV-REQUIMTE (UIDB/50006/2020). We also acknowledge FCT/FSE for the financial support to Maria João Ferreira through a PhD grant (PD/BD/150363/2019). We are grateful to Horta dos Peixinhos for their help and support during sampling and seed collection. We also thank Glória Pinto for her collaboration providing us the use of the growth chambers during the final months of the experiment and Enrique Mateos-Naranjo and Jennifer Mesa-Marín of the Departamento de Biología Vegetal y Ecología, the University of Sevilla for their advice regarding the growth of salicornia plants in greenhouse conditions.

Keywords: halophytes, PGPB, rhizosphere engineering, biofertilizers, primary metabolite profiling, plant inoculation, Salicornia ramosissima

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Authors: Armenio Lopes, Carlos Ferreira

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In the specific context of the Iberian Union between 1580 and 1640, characteristics emerged in Portuguese architecture that stood out from the main architectural production of the period. Recognised and identified aspects that had begun making their appearance decades before (1521) became significantly more marked during the Hapsburg-Spanish occupation. Distinctive even from the imperialist language of Spain, this trend would endure even after the restoration of independence (1706), continuing through to the start of the age of absolutism. Or perhaps not. This trend, recognised as Plain Style (Kubler), associated with a certain scarcity of resources, involved a certain formal and decorative simplification, as well as a particular set of conventions that would subsequently mark the landscape. This expression could also be seen as a means of asserting a certain spirit of independence as the Iberian Union breathed its last. The image of a simple, bare-bones architecture with purer design lines is associated by various authors –most notably Kubler– with the narratives of modernism, to whose principles it is similar, in a context-specific to the period. There is a contrast with some of the exuberance of the baroque or its expression in the Manueline period, in a similar fashion to modernism's responses to nineteenth-century eclecticism. This assertion and practice of simple architecture, drafted from the interpretation of the treaties, and highlighting a certain classical inspiration, was to become a benchmark in the theory of architecture, spanning the Baroque and Mannerism, until achieving contemporary recognition within certain originality and modernity. At a time when the baroque and its scenography became generally very widespread, it is important also to recognise the role played by plain style architecture in the construction of a rather complex and contradictory waterfront landscape, featuring promises of exuberance and more discrete practices.

Keywords: Carlos Mardel, Lisbon's waterfront, plain style, urban image on the waterfront

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Authors: Susana P. Dias, Andrea Gomes, Fernanda M. Ferreira, Marta F. Henriques

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Thymus mastichina L. is an endogenous medicinal and aromatic plant of the Mediterranean flora. It has been used empirically over the years as a natural preservative in food. Nowadays, the antimicrobial activity of its bioactive compounds, such as essential oils and extracts, has been well recognized. The main purpose of this study was to evaluate the antimicrobial effect of Thymus mastichina ethanolic and aqueous extracts on pathogens and spoilage microorganisms present in cheese during ripening. The effect that the extract type and its concentration has on the development of Staphylococcus aureus, Escherichia coli, and Yarrowia lipolytica populations during 24 hours, was studied 'in vitro' using appropriate culture media. The results achieved evidenced the antimicrobial activity of T. mastichina extracts against the studied strains, and the concentration of 2 mg/mL (w/v) was selected and used directly on the cheese surface during ripening. In addition to the microbiological evaluation in terms of total aerobic bacteria, Enterobacteriaceae, yeasts (particularly Y. lipolytica) and molds, the treated cheeses physicochemical evaluation (humidity, aw, pH, colour, and texture) was also performed. The results were compared with cheeses with natamicyn (positive control) and without any treatment (negative control). The physicochemical evaluation showed that the cheeses treated with ethanolic extract of Thymus mastichina, except the fact that they lead to a faster water loss during ripening, did not present considerable differences when compared to controls. The study revealed an evident antimicrobial power of the extracts, although less effective than the one shown by the use of natamycin. For this reason, the improvement of the extraction methods and the adjustment of the extract concentrations will contribute to the use of T. mastichina as a healthier and eco-friendly alternative to natamycin, that is also more attractive from an economic point of view.

Keywords: antimicrobial activity, cheese, ethanolic extract, Thymus mastichina

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Authors: Adriane Amend, Leidiene Ferreira Santos, Daniella Pires Nunes

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The experience of assuming or caring for older persons dependents by relatives is a complex task that encompasses or affective involvement, the demand for technical activities and or psychological support. It would be necessary to understand the situations related to the caregiver, the person and the environment, which help the family difficulty, as a caregiver to lead this role. Objective: To identify the forces that drive and restrict the care process of family caregivers of the older adults. Method: Descriptive and exploratory research, with a qualitative approach, which has as a reference the Force Field Theory. Five family caregivers of older adult’s dependents residing in the city of Palmas, Tocantins, Brazil will participate. The data were collected from December 2021 to February 2022, through a semi-structured individual interview, and submitted to content analysis. Results: As forces that drive or process of caring for family caregivers were: the account of compassionate attitudes and patience of the caregiver (I); to the collaboration of the other person to the care and to the body structure of the same (Other); and the supports of other people not cared for and structural, such as adaptations in the room, read and bathroom, as in the presence of air conditioners (Environment). Among the restrictive forces of care we mention difficulties in delegating care to another person, or stress of care and other personal demands (I); imposition of the older person about care and e a transfer from bed to hip (Other); e lack of accessibility of the house and absence of air conditioning and hospital bed (Environment). Conclusion: The results show that there are driving forces with the caregiver's attitude and feelings, a bond as an idol and support for the caregiver and the environment. On the other hand, conflicting ties, absence of physical structure and daily and continuous care shifts, can significantly compromise well-being or the cycle of older adult, caregiver and care.

Keywords: caregivers, frail elderly, perception, geriatric nursing

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Authors: Ana L. Martins, Rita S. Saraiva, João C. Ferreira

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Much effort has been placed on the assessment of perceived service quality. Several models can be found in literature, but these are mainly focused on business-to-consumer (B2C) relationships. Literature on how to assess perceived quality in business-to-business (B2B) contexts is scarce both conceptually and in terms of its application. This research aims at filling this gap in literature by applying INDSERV to a case study situation. Under this scope, this research aims at analyzing the adequacy of the proposed assessment tool to other context besides the one where it was developed and by doing so analyzing the perceive quality of the advertisement service provided by a specific television network to its B2B customers. The INDSERV scale was adopted and applied to a sample of 33 clients, via questionnaires adapted to interviews. Data was collected in person or phone. Both quantitative and qualitative data collection was performed. Qualitative data analysis followed content analysis protocol. Quantitative analysis used hypotheses testing. Findings allowed to conclude that the perceived quality of the television service provided by television network is very positive, being the Soft Process Quality the parameter that reveals the highest perceived quality of the service as opposed to Potential Quality. To this end, some comments and suggestions were made by the clients regarding each one of these service quality parameters. Based on the hypotheses testing, it was noticed that only advertisement clients that maintain a connection to the television network from 5 to 10 years do show a significant different perception of the TV advertisement service provided by the company in what the Hard Process Quality parameter is concerned. Through the collected data content analysis, it was possible to obtain the percentage of clients which share the same opinions and suggestions for improvement. Finally, based on one of the four service quality parameter in a B2B context, managerial suggestions were developed aiming at improving the television network advertisement perceived quality service.

Keywords: B2B, case study, INDSERV, perceived service quality

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Authors: Liliane Samara Ferreira Leite, Francys Kley Vieira Moreira, Luiz Henrique Capparelli Mattoso

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The increasing environmental concern regarding the plastic pollution worldwide has stimulated the development of low-cost biodegradable materials. Proteins are renewable feedstocks that could be used to produce biodegradable plastics. Gelatin, for example, is a cheap film-forming protein extracted from animal skin and connective tissues of Brazilian Livestock residues; thus it has a good potential in low-cost biodegradable plastic production. However, gelatin plastics are limited in terms of mechanical and barrier properties. Cellulose nanocrystals (CNC) are efficient nanofillers that have been used to extend physical properties of polymers. This work was aimed at evaluating the reinforcing efficiency of CNC on gelatin films. Specifically, we have employed the continuous casting as the processing method for obtaining the gelatin/CNC bionanocomposites. This required a first rheological study for assessing the effect of gelatin-CNC and CNC-CNC interactions on the colloidal state of the aqueous bionanocomposite formulations. CNC were isolated from eucalyptus pulp by sulfuric acid hydrolysis (65 wt%) at 55 °C for 30 min. Gelatin was solubilized in ultra-pure water at 85°C for 20 min and then mixed with glycerol at 20 wt.% and CNC at 0.5 wt%, 1.0 wt% and 2.5 wt%. Rotational measurements were performed to determine linear viscosity (η) of bionanocomposite solutions, which increased with increasing CNC content. At 2.5 wt% CNC, η increased by 118% regarding the neat gelatin solution, which was ascribed to percolation CNC network formation. Storage modulus (G’) and loss modulus (G″) further determined by oscillatory tests revealed that a gel-like behavior was dominant in the bionanocomposite solutions (G’ > G’’) over a broad range of temperature (20 – 85 °C), particularly at 2.5 wt% CNC. These results confirm effective interactions in the aqueous gelatin-CNC bionanocomposites that could substantially increase the physical properties of the gelatin plastics. Tensile tests are underway to confirm this hypothesis. The authors would like to thank the Fapesp (process n 2016/03080-3) for support.

Keywords: bionanocomposites, cellulose nanocrystals, gelatin, viscoelastic characterization

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Authors: Adriana Duarte, Sandra Sampaio, Catia Ferreira, Jaime I. N. R. Gomes

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With the fashion of faded worn garments the textile industry has moved from indigo and pigments to dyes that are fixed by cationization, with products that can be toxic, and that can show this effect after washing down the dye with friction and/or treating with enzymes in a subsequent operation. Increasingly they are treated with bleaches, such as hypochlorite and permanganate, both toxic substances. An alternative process is presented in this work for both garment and jet dyeing processes, without the use of pre-cationization and the alternative use of “particle-dyes”. These are hybrid products, made up by an inorganic particle and an organic dye. With standard soluble dyes, it is not possible to avoid diffusion into the inside of the fiber unless using previous cationization. Only in this way can diffusion be avoided keeping the centre of the fibres undyed so as to produce the faded effect by removing the surface dye and showing the white fiber beneath. With “particle-dyes”, previous cationization is avoided. By applying low temperatures, the dye does not diffuse completely into the inside of the fiber, since it is a particle and not a soluble dye, being then able to give the faded effect. Even though bleaching can be used it can also be avoided, by the use of friction and enzymes they can be used just as for other dyes. This fashion brought about new ways of applying reactive dyes by the use of previous cationization of cotton, lowering the salt, and temperatures that reactive dyes usually need for reacting and as a side effect the application of a more environmental process. However, cationization is a process that can be problematic in applying it outside garment dyeing, such as jet dyeing, being difficult to obtain level dyeings. It also should be applied by a pad-fix or Pad-batch process due to the low affinity of the pre-cationization products making it a more expensive process, and the risk of unlevelness in processes such as jet dyeing. Wit particle-dyes, since no pre-cationizartion is necessary, they can be applied in jet dyeing. The excess dye is fixed by a fixing agent, fixing the insoluble dye onto the surface of the fibers. By applying the fixing agent only one to 1-3 rinses in water at room temperature are necessary, saving water and improving the washfastness.

Keywords: denim, garment dyeing, worn look, eco-fashion

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Authors: Nádia Pereira, Paula Ferreira, Sofia Francisco, Sofia Oliveira, Sidclay Souza, Paula Paulino, Ana Margarida Veiga Simão

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Social media has progressed into a platform for hate speech among its users, and thus, there is an increasing need to develop automatic detection classifiers of offense and conflicts to help decrease the prevalence of such incidents. Online communication can be used to intentionally harm someone, which is why such classifiers could be essential in social networks. A possible application of these classifiers is the automatic detection of cyberbullying. Even though identifying the aggressive language used in online interactions could be important to build cyberbullying datasets, there are other criteria that must be considered. Being able to capture the language, which is indicative of the intent to harm others in a specific context of online interaction is fundamental. Offense and hate speech may be the foundation of online conflicts, which have become commonly used in social media and are an emergent research focus in machine learning and natural language processing. This study presents two Portuguese language offense-related datasets which serve as examples for future research and extend the study of the topic. The first is similar to other offense detection related datasets and is entitled Aggressiveness dataset. The second is a novelty because of the use of the history of the interaction between users and is entitled the Conflicts/Attacks dataset. Both datasets were developed in different phases. Firstly, we performed a content analysis of verbal aggression witnessed by adolescents in situations of cyberbullying. Secondly, we computed frequency analyses from the previous phase to gather lexical and linguistic cues used to identify potentially aggressive conflicts and attacks which were posted on Twitter. Thirdly, thorough annotation of real tweets was performed byindependent postgraduate educational psychologists with experience in cyberbullying research. Lastly, we benchmarked these datasets with other machine learning classifiers.

Keywords: aggression, classifiers, cyberbullying, datasets, hate speech, machine learning

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Authors: A. Camões, M. Ferreira Dias, M. Amorim

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In recent years, the social economy is increasingly seen as a viable means to address social problems. Social enterprises, as well as public projects and initiatives targeted to meet social purposes, offer organizational models that assume heterogeneity, flexibility and adaptability to the ‘real world and real problems’. Despite the growing popularity of social initiatives, decision makers still face a paucity in what concerns the available models and tools to adequately assess its sustainability, and its impacts, notably the nature of its contribution to economic growth. This study was carried out at the local level, by analyzing the social impact initiatives and projects promoted by the Municipality of Albergaria-a-Velha (Câmara Municipal de Albergaria-a-Velha -CMA), a municipality of 25,000 inhabitants in the central region of Portugal. This work focuses on the challenges related to the qualifications and employability of citizens, which stands out as one of the key concerns in the Portuguese economy, particularly expressive in the context of small-scale cities and inland territories. The study offers a characterization of the Municipality, its socio-economic structure and challenges, followed by an exploratory analysis of multiple sourced data, collected from the CMA's documental sources as well as from privileged informants. The purpose is to conduct detailed analysis of the CMA's social projects, aimed at characterizing its potential impact for the model of qualifications and employability of the citizens of the Municipality. The study encompasses a discussion of the socio-economic profile of the municipality, notably its asymmetries, the analysis of the social projects and initiatives, as well as of data derived from inquiry actors involved in the implementation of the social projects and its beneficiaries. Finally, the results obtained with the Better Life Index will be included. This study makes it possible to ascertain if what is implicit in the literature goes to the encounter of what one experiences in reality.

Keywords: measurement, municipalities, social economy, social impact

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Authors: Maria Lucia G. Lourenco, Viviane Y. Hibaru, Keylla H. N. P. Pereira, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Isosporiasis is an affliction caused by coccidial protozoa belonging to genera Isospora spp. or Cystoisospora spp., which may parasitize the small and large intestines of dogs, of which neonates and young animals present higher risk of infection. This study aims at reporting a case of isosporiasis in neonate Pitbull dogs, as well as the diagnosis and treatment. Seven Pitbull puppies were admitted to the São Paulo State University (UNESP) Veterinary Hospital, Botucatu, São Paulo, Brazil, with history of yellowish diarrhea without mucus or blood for the past two days. The animals were five days old. The history of the mother, a primiparous two-year-old, revealed that she was properly vaccinated, not de-wormed and did not present diarrhea. The clinical examination revealed that the neonates weighted between 308 and 360 grams, and presented normal reflexes, moderate dehydration, body temperatures between 36.8 and 37.2 ºC, blood sugar between 103 and 124 mg/dL and normal appetite. A full blood count and a parasitology assay were performed to aid in the diagnosis. The full blood count detected eosinophilia, without any other relevant alterations. The parasitology assay (Willis-Molly & Faust) revealed the presence of Cystoisospora spp. The treatment was instituted with heated fluid therapy with Ringer’s Lactate (4 mL/100 g, subcutaneous) and antibiotic therapy with sulfamethoxazole-trimethoprim (15 mg/kg, orally) every 12 hours for ten days. The mother and other dogs that came in contact with the newborns were also treated. The environment was disinfected for 10 minutes with 1.6% quaternary ammonium. After 10 days, the newborns presented normal clinical signs and no alterations in the full blood count. Isosporiasis is an affliction with high mortality rates in litters that should be diagnosed and treated as soon as possible to increase the survival rates in these patients.

Keywords: Cystoisospora spp., neonatal infection, puppies, diarrhea,

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Authors: Maria L. G. Lourenço, Keylla H. N. P. Pereira, Viviane Y. Hibaru, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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In veterinary medicine, the standard procedure during a caesarian section is clamping the umbilical cord immediately after birth. In human neonates, when the umbilical cord is kept intact after birth, blood continues to flow from the cord to the newborn, but this procedure may prove to be difficult in dogs due to the shorter umbilical cord and the number of newborns in the litter. However, a possible detachment of the placenta while keeping the umbilical cord intact may make the residual blood to flow to the neonate. This study compared the effects on neonatal vitality between clamping and no clamping the umbilical cord of dogs born through cesarean section, assessing them through Apgar and reflex scores. Fifty puppies delivered from 16 bitches were randomly allocated to receive clamping of the umbilical cord immediately (n=25) or to not receive the clamping until breathing (n=25). The neonates were assessed during the first five min of life and once again 10 min after the first assessment. The differences observed between the two moments were significant (p < 0.01) for both the Apgar and reflex scores. The differences observed between the groups (clamped vs. not clamped) were not significant for the Apgar score in the 1st moment (p=0.1), but the 2nd moment was significantly (p < 0.01) in the group not clamped, as well as significant (p < 0.05) for the reflex score in the 1st moment and 2nd moment (p < 0.05), revealing higher neonatal vitality in the not clamped group. The differences observed between the moments (1st vs. 2nd) of each group as significant (p < 0.01), revealing higher neonatal vitality in the 2nd moments. In the no clamping group, after removing the neonates together with the umbilical cord and the placenta, we observed that the umbilical cords were full of blood at the time of birth and later became whitish and collapsed, demonstrating the blood transfer. The results suggest that keeping the umbilical cord intact for at least three minutes after the onset breathing is not detrimental and may contribute to increase neonate vitality in puppies delivered by cesarean section.

Keywords: puppy vitality, newborn dog, cesarean section, Apgar score

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Authors: Maria L. G. Lourenço, Keylla H. N. P. Pereira, Viviane Y. Hibaru, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Anasarca is generalized congenital edema that is often lethal. The condition is transmitted hereditarily and is autosomal dominant, with a racial predisposition in French Bulldogs and English Bulldogs. This study aims at reporting a case of anasarca treatment in neonates. The fetuses of a one year and six months old, primiparous English Bulldog mother were diagnosed with anasarca during an ultrasound examination performed at the 55th day of pregnancy and, therefore, an elective cesarean section was scheduled to prevent fetal dystocia. At birth, all puppies presented anasarca, and one of the six was stillborn. The newborns presented cyanosis, dyspnea, bradycardia, absent reflexes, low vitality scores (3/10), and hypothermia ( < 32ºC). The weight of the puppies at the time of birth varied between 347 and 373 grams, about 100 grams above the average weight estimated for the breed. Immediate neonatal care was applied with oxygen therapy via a mask, aminophylline (0.2 ml/100 g/PV/sublingual), and slow heating. After 10 minutes, there was a significant improvement in the neonatal parameters. The anasarca was treated with the drug furosemide, administered subcutaneously, at a dose of 0.2 mg per 100 grams of weight, every three hours. The stimulation for urination of newborns was performed every 30 minutes, and weight loss was monitored every 30 minutes. Five grams of potassium chloride were administered orally for every 30 grams of weight loss to counterbalance the loss of potassium caused by the diuretic medication. After 15 hours, the neonates reached the ideal weight for the breed, around 209 to 230 grams. In total, four neonates received five doses of furosemide, while one received six doses. The puppies are currently ten months old, healthy and neutered. Anasarca should not be ignored and is considered potentially lethal and an indication for euthanasia in all cases. Early intervention is of utmost importance for the survival of these patients.

Keywords: Walrus syndrome, congenital edema, water puppy syndrome, puppies

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Authors: Jéssica Ferreira Rodrigues, Júlio Silvio De Sousa Bueno Filho, Vanessa Rios De Souza, Ana Carla Marques Pinheiro

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This study aimed to evaluate the implications of the block size and testing order on efficiency and precision of preference estimation for Dulce de leche samples. Efficiency was defined as the inverse of the average variance of pairwise comparisons among treatments. Precision was defined as the inverse of the variance of treatment means (or effects) estimates. The experiment was originally designed to test 16 treatments as a series of 8 Sudoku 16x16 designs being 4 randomized independently and 4 others in the reverse order, to yield balance in testing order. Linear mixed models were assigned to the whole experiment with 112 testers and all their grades, as well as their partially balanced subgroups, namely: a) experiment with the four initial EU; b) experiment with EU 5 to 8; c) experiment with EU 9 to 12; and b) experiment with EU 13 to 16. To record responses we used a nine-point hedonic scale, it was assumed a mixed linear model analysis with random tester and treatments effects and with fixed test order effect. Analysis of a cumulative random effects probit link model was very similar, with essentially no different conclusions and for simplicity, we present the results using Gaussian assumption. R-CRAN library lme4 and its function lmer (Fit Linear Mixed-Effects Models) was used for the mixed models and libraries Bayesthresh (default Gaussian threshold function) and ordinal with the function clmm (Cumulative Link Mixed Model) was used to check Bayesian analysis of threshold models and cumulative link probit models. It was noted that the number of samples tested in the same session can influence the acceptance level, underestimating the acceptance. However, proving a large number of samples can help to improve the samples discrimination.

Keywords: acceptance, block size, mixed linear model, testing order, testing order

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Authors: Eliane Ferreira da Silva

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This paper aims to mobilize and sensitize public administration leaders to good practices and encourage investment in the PV system in Brazil. It presents a case study methodology for dimensioning the PV system in the roofs of the public buildings of the Esplanade of the Ministries, Brasilia, capital of the country, with predefined resources, starting with the Sustainable Esplanade Project (SEP), of the exponential growth of photovoltaic solar energy in the world and making a comparison with the solar power plant of the Ministry of Mines and Energy (MME), active since: 6/10/2016. In order to do so, it was necessary to evaluate the energy efficiency of the buildings in the period from January 2016 to April 2017, (16 months) identifying the opportunities to reduce electric energy expenses, through the adjustment of contracted demand, the tariff framework and correction of existing active energy. The instrument used to collect data on electric bills was the e-SIC citizen information system. The study considered in addition to the technical and operational aspects, the historical, cultural, architectural and climatic aspects, involved by several actors. Identifying the reductions of expenses, the study directed to the following aspects: Case 1) economic feasibility for exchanges of common lamps, for LED lamps, and, Case 2) economic feasibility for the implementation of photovoltaic solar system connected to the grid. For the case 2, PV*SOL Premium Software was used to simulate several possibilities of photovoltaic panels, analyzing the best performance, according to local characteristics, such as solar orientation, latitude, annual average solar radiation. A simulation of an ideal photovoltaic solar system was made, with due calculations of its yield, to provide a compensation of the energy expenditure of the building - or part of it - through the use of the alternative source in question. The study develops a methodology for public administration, as a major consumer of electricity, to act in a responsible, fiscalizing and incentive way in reducing energy waste, and consequently reducing greenhouse gases.

Keywords: energy efficiency, esplanade of ministries, photovoltaic solar energy, public buildings, sustainable building

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Authors: C. Miranda, R. Soares, S. Cunha, L. Ferreira, G. Igrejas, P. Poeta

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Different animal species, including ornamental animals, are reported as potential reservoirs of antibiotic resistance genes. Consequently, these resistances can be disseminated in the environment and transferred to humans. Moreover, multidrug-resistant bacteria reduce the efficacy of antibiotics, as the case of vancomycin-resistant enterococci. Enterococcus faecalis and E. faecium are described as the main nosocomial pathogens. In this line, the aim of this study was to characterize resistance and virulence genes of enterococci species isolated from samples of food supplied to ornamental animals during 2020. The 29 enterococci isolates (10 E. faecalis and 19 E. faecium) were tested for the presence of the resistance genes for the following antibiotics: erythromicyn (ermA, ermB and ermC), tetracycline (tetL, tetM, tetK and tetO), quinupristin/dalfopristin (vatD and vatE), gentamicin (aac(6’)-aph(2’’)-Ia), chloramphenicol (catA), streptomycin (ant(6)-Ia) and vancomycin (vanA and vanB). The same isolates were also tested for 10 virulence factors genes (esp, ace, gelE, agg, fsr, cpd, cylA, cylB, cylM and cylLL). The resistance and virulence genes were performed by PCR, using specific primers and conditions. Negative and positive controls were used in all PCR assays. The most prevalent resistance genes detected in both enterococci species were ermB (n=15, 52%), ermC (n=7, 24%), tetK (n=8, 28%) and vatE (n=4, 14%). Resistance genes for vancomycin were found in ten (34%) E. faecalis and ten (34%) E. faecium isolates. Only E. faecium isolates showed the presence of ermA (n=2, 7%), tetL (n=13, 45%) and ant(6)-Ia gene (n=4, 14%). A total of nine (31%) enterococci isolates were classified as multidrug-resistant bacteria (3 E. faecalis and 6 E. faecium). In three E. faecalis and one E. faecium were not detected resistance genes. The virulence genes detected in both species were agg (n=6, 21%) and cylLL (n=11, 38%). In general, each isolate showed only one of these virulence genes. Five E. faecalis and eleven E. faecium isolates were negative for all analyzed virulence genes. These preliminary results showed the presence of multidrug-resistant enterococci in food supplied to ornamental animals, in particular vancomycin-resistant enterococci. This genotypic characterization reinforces the relevance to public health in the control of antibiotic-resistant bacteria.

Keywords: antibiotic resistance, enterococci, feed, ornamental animals

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Authors: Hamza I. Isa, Gezina C. H. Ferreira, Jan E. Crafford, Christoffel J. Botha

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Moraea pallida Bak. (yellow tulip) poisoning is the most important plant-induced cardiac glycoside toxicosis in South Africa. Cardiac glycoside poisonings collectively account for about 33 and 10 % mortalities due to plants, in large and small stock respectively, in South Africa. The toxic principle is 1α, 2α-epoxyscillirosidine, a bufadienolide. The aim of the study was to investigate the potential to develop a vaccine against epoxyscillirosidine. Epoxyscillirosidine and the related bufadienolides proscillaridin and bufalin, which are commercially available, were conjugated to the carrier proteins [Hen ovalbumin (OVA), bovine serum albumin (BSA) and keyhole limpet haemocyanin (KLH)], rendering them immunogenic. Adult male New Zealand White rabbits were immunized. In Trials 1 and 2, rabbits (n=6) were, each assigned to two groups. Experimental animals (n=3; n=4) were vaccinated with epoxyscillirosidine-OVA conjugate, while the control (n=3; n=2) were vaccinated with OVA, using Freund’s complete and incomplete and Montanide adjuvants, for Trials 1 and 2, respectively. In Trial 3, rabbits (n=15), randomly allocated to 5 equal groups (I, II, III, IV and V), were vaccinated with proscillaridin-BSA, bufalin-BSA, epoxyscillirosidine-KLH, epoxyscillirosidine-BSA conjugates, and BSA respectively, using Montanide as adjuvant. Vaccination was on Days 0, 21 and 42. Additional vaccinations were done on Day 56 and 63 for Trial 1. Vaccination was by intradermal injection of 0.4 ml of the immunogen (4 mg/ml [Trial 1] and 8 mg/ml for Trials 2 and Trial 3, respectively). Blood was collected pre-vaccination and at 3 week intervals following each vaccination. Antibody response was determined using an indirect ELISA. There was poor immune response associated with the dose (0.4 mg per rabbit) and adjuvant used in Trial 1. Antibodies were synthesized against the conjugate administered in Trial 2. For Trail 3, antibodies against the immunogens were successfully raised in rabbits with epoxyscillirosidine-KLH inducing the highest immune response. The antibodies raised against proscillaridin and bufalin cross-reacted with epoxyscillirosidine when used as antigen in the ELISA. The study successfully demonstrated the synthesis of antibodies against the bufadienolide conjugates administered. The cross-reactivity of proscillaridin and bufalin with epoxyscillirosidine could potentially be utilized as alternative to epoxyscillirosidine in future studies to prevent yellow tulp poisoning by vaccination.

Keywords: antibodies , bufadienolides, cross-reactivity, epoxyscillirosidine, Moraea pallida, poisoning

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