Sensitive bi-enzymatic biosensor based on polyphenoloxidases–gold nanoparticles–chitosan hybrid film–graphene doped carbon paste electrode for carbamates detection

A bi-enzymatic biosensor (LACC–TYR–AuNPs–CS/GPE) for carbamates was prepared in a single step by electrodeposition of a hybrid film onto a graphene doped carbon paste electrode (GPE). Graphene and the gold nanoparticles (AuNPs) were morphologically characterized by transmission electron microscopy, X-ray photoelectron spectroscopy, dynamic light scattering and laser Doppler velocimetry. The electrodeposited hybrid film was composed of laccase (LACC), tyrosinase (TYR) and AuNPs entrapped in a chitosan (CS) polymeric matrix. Experimental parameters, namely graphene redox state, AuNPs:CS ratio, enzymes concentration, pH and inhibition time were evaluated. LACC–TYR–AuNPs–CS/GPE exhibited an improved Michaelis–Menten kinetic constant (26.9 ± 0.5 M) when compared with LACC–AuNPs–CS/GPE (37.8 ± 0.2 M) and TYR–AuNPs–CS/GPE (52.3 ± 0.4 M). Using 4-aminophenol as substrate at pH 5.5, the device presented wide linear ranges, low detection limits (1.68×10− 9 ± 1.18×10− 10 – 2.15×10− 7 ± 3.41×10− 9 M), high accuracy, sensitivity (1.13×106 ± 8.11×104 – 2.19×108 ± 2.51×107 %inhibition M− 1), repeatability (1.2–5.8% RSD), reproducibility (3.2–6.5% RSD) and stability (ca. twenty days) to determine carbaryl, formetanate hydrochloride, propoxur and ziram in citrus fruits based on their inhibitory capacity on the polyphenoloxidases activity. Recoveries at two fortified levels ranged from 93.8 ± 0.3% (lemon) to 97.8 ± 0.3% (orange). Glucose, citric acid and ascorbic acid do not interfere significantly in the electroanalysis. The proposed electroanalytical procedure can be a promising tool for food safety control.

Utilization of food industry wastes for the production of zero-valent iron nanoparticles

The proper disposal of the several types of wastes produced in industrial activities increases production costs. As a consequence, it is common to develop strategies to reuse these wastes in the same process and in different processes or to transform them for use in other processes. This work combines the needs for new synthesis methods of nanomaterials and the reduction of production cost using wastes from citrine juice (orange, lime, lemon and mandarin) to produce a new added value product, green zero-valent iron nanoparticles that can be used in several applications, including environmental remediation. The results indicate that extracts of the tested fruit wastes (peel, albedo and pulp fractions) can be used to produce zero-valent iron nanoparticles (nZVIs). This shows that these wastes can be an added value product. The resulting nZVIs had sizes ranging from 3 up to 300 nm and distinct reactivities (pulp > peel > albedo extracts). All the studied nanoparticles did not present a significant agglomeration/settling tendency when compared to similar nanoparticles, which indicates that they remain in suspension and retain their reactivity.

25 kV bipolar solid-state Marx generator for industrial food applications

The preliminary results from a bipolar industrial solidstate based Marx generator, developed for the food industry, capable of delivering 25 kV/250 A positive and negative pulses with 12 kW average power, are presented and discussed. This modular topology uses only four controlled switches per cell, 27 cells in total that can be charged up to 1000V each, the two extra cells are used for droop compensation. The triggering signals for all the switches are generated by a FPGA. Considering that biomaterials are similar to resistive type loads, experimental results from this new bipolar 25 kV modulator into resistive loads are presented and discussed.

Salmonella serovars in food poisoning episodes recorded in Brazil from 1982 to 1991

The Salmonella serovars involved in 25 food poisoning episodes which occurred in the Southeast and South of Brazil from 1982 to 1991 were identified. The most frequently detected serotype was S. Typhimurium (13/25, 52%), and the food most frequently involved in the transmission of Salmonella was homemade mayonnaise. The need to set up a permanent program of epidemiologic alert for food poisoning is emphasized.

Intestinal parasites in school food handlers in the city of Uberlândia, Minas Gerais, Brazil

In order to verify the presence of intestinal parasites in food handlers, stool samples were collected from 104 cooks and their helpers that were working in food preparation in 20 public elementary schools, in various areas of the city of Uberlândia, Minas Gerais, Brazil. The samples were collected during the months of November and December, 1988, in plastic flasks containing a 10% formaldehyde solution and processed by the Hoffmann, Pons & Janer method. The sediment was examined using triplicate slides. All individuals were females aged between 24 to 69 years. Intestinal parasites were found in 85.0% of the studied schools and 47.1% of the studied food handlers (cooks and helpers) were found to be positive. Among the 49 infected food handlers, 32 (65.3%) carried a single parasite and 17 (34.7%) carried two parasites. The following intestinal parasites were found: Giardia lamblia (21.1%), Entamoeba coli (21.1%), hookworms (9.6%), Ascaris lumbricoides (5.8%), Entamoeba histolytica (2.9%), Hymenolepis nana (1.9%), Strongyloides stercoralis (1.0%). These data emphasize the need for a rigid semi-annual control in all school food handlers, including diagnosis, specific treatment and orientation about the mechanisms of transmission of the intestinal parasites.

Freshwater arsenic detoxification through selenium-enriched food supplements. A proteomic approach

Dissertação apresentada para a obtenção do grau de doutor em Bioquímica pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Stress Ratio Effects on the fatigue Properties of Biaxially Loaded Tubular FRP-Specimens – Experimental Study

In this work tubular fiber reinforced specimens are tested for fatigue life. The specimens are biaxially loaded with tension and shear stresses, with a load angle β of 30° and 60° and a load ratio of R=0,1. There are many factors that affect fatigue life of a fiber reinforced material and the main goal of this work is to study the effects of load ratio R by obtaining S-N curves and compare them to the previous works (1). All the other parameters, such as specimen production, fatigue loading frequency and temperature, will be the same as for the previous tests. For every specimen, stiffness, temperature of the specimen during testing, crack counting and final fracture mode are obtained. Prior to testing, a study if the literature regarding the load ratio effects on composites fatigue life and with that review estimate the initial stresses to be applied in testing. In previous works (1) similar specimens have only been tested for a load ratio of R=-1 and therefore the behaviour of this tubular specimens for a different load ratio is unknown. All the data acquired will be analysed and compared to the previous works, emphasizing the differences found and discussing the possible explanations for those differences. The crack counting software, developed at the institute, has shown useful before, however different adjustments to the software parameters lead to different cracks numbers for the same picture, and therefore a better methodology will be discussed to improve the crack counting results. After the specimen’s failure, all the data will be collected and stored and fibre volume content for every specimen is also determinate. The number of tests required to make the S-N curves are obtained according to the existent standards. Additionally are also identified some improvements to the testing machine setup and to the procedures for future testing.

Hepatocellular carcinoma in Brazil: report of a national survey (Florianópolis, SC, 1995)

In order to investigate epidemiological aspects of hepatocellular carcinoma (HCC) in Brazil, basic informations about cases diagnosed from January 1992 to December 1994 were requested to several medical centers of different Brazilian States. A simple questionnaire included age, sex, alcohol abuse (over 80g/day), associated liver cirrhosis, persistent HBV infection (HBsAg), HCV infection (anti-HCV) and serum levels of alpha fetoprotein. 287 cases, over 16 years old, from 19 medical centers of 8 States (Pará, Bahia, Minas Gerais, Espirito Santo, Rio de Janeiro, São Paulo, Paraná and Rio Grande do Sul) were analysed. The results showed: (a) Mean age was 56.3 ± 14.4 for men and 54.7 ± 16.8 yr for women and the male/female ratio was 3.4:1. (b) 69.6% were caucasians, 21.8% mullatoes, 4.8% orientals and 3.7% blacks. (c) HBsAg (+) in 77/236 cases (41.6%) without differences between males and females. (d) Anti-HCV (+) in 52/193 cases (26.9%). (e) 7/180 cases were positive both for HBsAg and anti-HCV (3.8%). (f) There was chronic alcoholism in 88/235 cases (37%). (g) HCC was found in cirrhotic livers in 71.2% of 202 cases in which the presence or absence of cirrhosis was reported. (h) Alpha-fetoprotein above 20 ng/ml was found in 124/172 cases (72%) and above 500 ng/ml only in 40 cases (23.2%). These results showed that the HCC in Brazil has an intermediate epidemiological pattern as compared to those from areas of low and high incidence of the tumor. In spite of the high frequency of the association of HCC with the HBV and/or HCV infections, 42% of 180 cases were negative both for HBsAg and anti-HCV, indicating the possible role of other etiological factors. The comparison of data from different States showed some regional differences: higher frequency of associated HBsAg in Pará, Bahia, Minas Gerais and Espírito Santo, higher frequency of associated HCV infection in Rio de Janeiro, São Paulo and States of the Southern region and low frequency of associated liver cirrhosis in Salvador and Rio de Janeiro (55.5 and 50% respectively). Further investigation will be necessary to study the presence of other possible etiological factors as aflatoxins, suggested by the favourable climatic conditions for food contamination by fungi in the majority Brazilian regions

Vitamin D Intake and Cardiometabolic: Risk Factors in Adolescents

Background: A growing body of research suggests that vitamin D might play an important role in overall health. No data exist on vitamin D intake for the Azorean adolescent population. The purpose of this study was to assess vitamin D intake and investigate a possible association between vitamin D intake and cardiometabolic risk factors in Azorean adolescents. Methods: A cross-sectional school-based study was conducted on 496 adolescents (288 girls) aged 15–18 years from the Azorean Islands, Portugal. Anthropometric measurements (waist circumference and height), blood pressure (systolic), and plasma biomarkers [fasting glucose, insulin, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and triglycerides (TGs)] were measured to assess metabolic risk. Homeostasis model assessment (HOMA), TC-to-HDL-C ratio, and waist-to-height ratio were calculated. For each of these variables, a Z-score was computed by age and sex. A metabolic risk score was constructed by summing the Zscores of all individual risk factors. High risk was considered when the individual had ‡ 1 standard deviation(SD) of this score. Vitamin D intake was assessed with a semiquantitative food frequency questionnaire. Participants were classified into quartiles of vitamin D intake. Logistic regression was used to determine odds ratios for high cardiometabolic risk scores after adjusting for total energy intake, pubertal stage, fat mass percentage, and cardiorespiratory fitness. Results: Mean (SD) vitamin D intake was 5.8 (6.5) mg/day, and 9.1% of Azorean adolescents achieved the estimated average requirement of vitamin D (10 mg/day or 400 IU). Logistic regression showed that the odds ratio for a high cardiometabolic risk score was 3.35 [95% confidence interval (CI) 1.28–8.75] for adolescents in the lowest vitamin D intake quartile in comparison with those in the highest vitamin D intake quartile, even after adjustment for confounders. Conclusion: A lower level of vitamin D intake was associated with worse metabolic profile among Azorean adolescents.

Intake of milk, but not total dairy, yogurt, or cheese, is negatively associated with the clustering of cardiometabolic risk factors in adolescents

Epidemiologic studies have reported an inverse association between dairy product consumption and cardiometabolic risk factors in adults, but this relation is relatively unexplored in adolescents. We hypothesized that a higher dairy product intake is associated with lower cardiometabolic risk factor clustering in adolescents. To test this hypothesis, a cross-sectional study was conducted with 494 adolescents aged 15 to 18 years from the Azorean Archipelago, Portugal. We measured fasting glucose, insulin, total cholesterol, high-density lipoprotein cholesterol, triglycerides, systolic blood pressure, body fat, and cardiorespiratory fitness. We also calculated homeostatic model assessment and total cholesterol/high-density lipoprotein cholesterol ratio. For each one of these variables, a z score was computed using age and sex. A cardiometabolic risk score (CMRS) was constructed by summing up the z scores of all individual risk factors. High risk was considered to exist when an individual had at least 1 SD from this score. Diet was evaluated using a food frequency questionnaire, and the intake of total dairy (included milk, yogurt, and cheese), milk, yogurt, and cheese was categorized as low (equal to or below the median of the total sample) or “appropriate” (above the median of the total sample).The association between dairy product intake and CMRS was evaluated using separate logistic regression, and the results were adjusted for confounders. Adolescents with high milk intake had lower CMRS, compared with those with low intake (10.6% vs 18.1%, P = .018). Adolescents with appropriate milk intake were less likely to have high CMRS than those with low milk intake (odds ratio, 0.531; 95% confidence interval, 0.302-0.931). No association was found between CMRS and total dairy, yogurt, and cheese intake. Only milk intake seems to be inversely related to CMRS in adolescents.

Association between dairy product intake and abdominal obesity in Azorean adolescents

BACKGROUND: Some studies have reported an inverse association between dairy product (DP) consumption and weight or fat mass loss. OBJECTIVES: The objective of our study was to assess the association between DP intake and abdominal obesity (AO) among Azorean adolescents. SUBJECTS/METHODS: This study was a cross-sectional analysis. A total of 903 adolescents (370 boys) aged 15--16 years was evaluated. Anthropometric measurements were collected (weight, height and waist circumference (WC)) and McCarthy’s cut-points were used to categorize WC. AO was defined when WC was X90th percentile. Adolescent food intake was assessed using a self-administered semiquantitative food frequency questionnaire and DP intake was categorized in o2 and X2 servings/day. Data were analyzed separately for girls and boys, and logistical regression was used to estimate the association between DPs and AO adjusting for potential confounders. RESULTS: The prevalence of AO was 54.9% (boys: 32.1% and girls: 70.7%, Po0.001). For boys and girls, DP consumption was 2.3±1.9 and 2.1±1.6 servings/day (P¼0.185), respectively. In both genders, the proportion of adolescents with WC o90th percentile was higher among individuals who reported a dairy intake of X2 servings/day compared with those with an intake o2 servings/day (boys: 71% vs 65% and girls: 36% vs 24%, Po0.05). After adjustments for confounders, two or more DP servings per day were a negative predictor of AO (odds ratio, 0.217; 95% confidence interval, 0.075 -- 0.633) only in boys. CONCLUSION: We found a protective association between DP intake and AO only in boys.

Estudo comparativo da utilização do biodiesel produzido a partir de óleos alimentares usados e do gasóleo numa frota industrial

A definição de teores mínimos de incorporação de biocombustíveis, constitui objeto de discussão entre grupos pro-desenvolvimento e ambientalistas. Esses últimos argumentam que as consequências da utilização desta fonte energética ainda são desconhecidas. Além disso, alegam que a produção de biocombustíveis é, em parte, responsável pelo aumento no preço dos alimentos, encoraja a conversão de florestas em monoculturas e conduz à exploração de trabalhadores em países em desenvolvimento (PEDs). Para responder à dependência energética dos combustíveis de origem fóssil, e ajudar a reduzir as emissões de gases com efeito de estufa, sobretudo no sector dos transportes, o biodiesel produzido a partir de óleos alimentares usados têm sido apontado como uma “solução verde” capaz de minimizar o problema das alterações climáticas e valorizar um resíduo, e simultaneamente conferir ao setor energético um pouco mais de independência. De forma a desmistificar e clarificar um pouco estas premissas, a presente dissertação pretende fazer um estudo de avaliação do impacto da utilização do biodiesel, nomeadamente no que diz respeito às emissões gasosas. Posteriormente realizou-se, tomando como referência uma pequena frota industrial existente, uma análise comparativa dos consumos e emissões dos principais poluentes decorrentes da utilização do biodiesel em diferentes percentagens de incorporação no gasóleo, comparativamente ao gasóleo puro. O trabalho culmina com uma abordagem técnica sobre o comportamento de um veículo equipado com um motor de ignição por compressão, utilizando como biocombustível o biodiesel.

Utilização da Nanotecnologia para o desenvolvimento sustentável na indústria agro-alimentar

O crescimento populacional esperado para os próximos anos conduzirá à necessidade de aumentar a produção agrícola de modo a satisfazer o aumento da procura. Nos últimos anos tem-se assistido a uma evolução tecnológica nos sistemas de produção que tem permitido aumentar a produtividade agrícola, por vezes à custa de elevados consumos de energia e com práticas nem sempre ambientalmente corretas. Os desafios que se colocam atualmente são no sentido de melhorar a conservação de recursos escassos, como o solo e a água, de aumentar a eficiência de uso de fatores de produção, de encontrar novas culturas, do desenvolvimento da biotecnologia, da diminuição dos consumos energéticos e de melhorar ainda mais as tecnologias associadas à produção. De maneira a responder aos desafios emergentes da procura por alimentos, da escassez de terrenos agrícolas aráveis bem como da existência de pragas de insetos e de ervas daninhas, os pesticidas tem vindo a ser usados com maior frequência, tendo-se assistido a uma contaminação dos solos e águas subterrâneas, causando deste modo um risco para a saúde dos seres vivos. Neste sentido, vários fabricantes de pesticidas estão a desenvolver novas formulações contendo pesticidas encapsulados em nanopartículas como modo de aumentar a sua solubilidade em água, biodisponibilidade, volatilidade, estabilidade e eficácia. tendo por objetivo um desenvolvimento sustentável. Neste trabalho, procedeu-se ao estudo do encapsulamento do herbicida Oxadiargil (5-terc-butil-3-[2,4-dicloro-5-(2-propiniloxi)fenil]-1,3,4-oxadiazol-2(3H)-ona) com a 2-hidroxipropil-β-ciclodextrina (HP-β-CD). O estudo da formação do complexo de inclusão Oxadiargil - HP-β-CD foi realizado em diferentes meios, água desionizada, tampão acetato pH = 3,46 e pH = 5,34 e tampão fosfato pH = 7,45, com o objetivo de determinar e comparar a sua constante de estabilidade. Verificou-se, em qualquer dos casos, a ocorrência de uma relação linear entre o aumento da solubilidade do Oxadiargil e o aumento da concentração de HP-β-CD, com um declive inferior a um, o que indicia a formação de um complexo na proporção estequiométrica de 1:1. Os resultados obtidos permitiram concluir que o processo de complexação Oxadiargil - HP-β-CD não é muito influenciado pela constituição e pelo pH do meio. De facto, as constantes de estabilidade obtidas para a água desionizada e soluções-tampão pH = 3,46, pH = 5,34 e pH = 7,45 foram de 919 ± 25, 685 ± 13, 623 ± 17 e 753 ± 9, respetivamente. A solubilidade do complexo obtido nos estudos realizados, em diferentes meios, é cerca de 23 a 32 vezes superior à observada para o Oxadiargil livre. De forma a caracterizar o complexo Oxadiargil - HP-β-CD procedeu-se à sua síntese utilizando o método de “kneading”. O composto obtido foi caracterizado por Ressonância Magnética Nuclear (RMN) tendo-se confirmado a formação de um complexo de inclusão na proporção estequiométrica de 1:1. O complexo obtido é mais solúvel e porventura mais estável quimicamente. O encapsulamento permite uma redução da aplicação dos pesticidas diminuindo assim os custos e o impacto negativo no ambiente. Com a nanotecnologia é possível a libertação controlada dos pesticidas, aumentando a sua eficácia e fornecendo os meios necessários para um desenvolvimento sustentável.

Botulism: a laboratory investigation on biological and food samples from cases and outbreaks in Brazil (1982-2001)

Laboratory investigation of botulism from 1982 to 2001 confirmed the occurrence of eight positive outbreaks/cases of botulism in Brazil. From those, type A botulism was observed in seven of them. Biological material of one case (serum and feces) was positive in the first step of the bioassay, but the amount of sample was not sufficient for typification. One of the outbreaks that occurred in 2001 was negative for botulinum toxin in samples of serum, gastric washing and feces, collected eight days before the onset of the symptoms in the affected person who was clinically diagnosed as presenting the disease. Other two cases presenting compatible clinical diagnoses presented negative results. However, in those cases, the collection of samples was (1) after antiserum administration or (2) later than eight days of the onset of symptoms. Investigation was performed by mouse bioassay, as described in the Compendium of Methods for the Microbiological Examination of Foods (compiled by American Public Health Association - APHA)11, using specific antiserum from Centers for Disease Control (CDC), USA.

Phage types of Salmonella Enteritidis isolated from clinical and food samples, and from broiler carcasses in Southern Brazil

272 isolates of Salmonella Enteritidis (111 isolated from frozen broiler chicken carcasses, 126 from human food and other biological materials involved in food poisoning outbreaks and 35 from different poultry materials) were selected for phage typing. From these, 111 were phage typed, 57.65% being classified as phage type 4, 32.43% as phage type 4a, 3.60% as phage type 6a and 0.90% as phage type 7, whereas 5.40% samples were not phage typeable. The predominance of phage type 4 is in agreement with the results published worldwide, and reinforces the need for studies related to the epidemiological meaning of these findings.