A fumonisina B1 (FB1) e uma micotoxina produzida pelo metabolismo secundário de espécies de Fusarium, principalmente F. verticillioides e F. proliferatum, os quais contaminam diversos alimentos antes e apos o processamento, sobretudo o milho e derivados, gerando graves problemas para a Saúde Pública e a qualidade dos alimentos. O objetivo deste trabalho foi avaliar a exposição humana a FB1 presente nos alimentos através da estimativa de ingestão da toxina na dieta e da análise de diferentes biomarcadores presentes em amostras de sangue, urina e cabelo. Além disso, foram investigados os efeitos da toxina através da avaliação de ácido fólico presentes em alimentos e em soro, e os níveis de uréia e creatinina presentes em soro. O estudo foi realizado em dois municípios dos Estados de São Paulo e Santa Catarina, cujos respectivos voluntários foram categorizados como de baixo consumo de derivados de milho (Grupo A, voluntários de Pirassununga/SP) e de alto consumo de derivados de milho (Grupo B, voluntários de Erval Velho/SC). As amostras de alimentos do Grupo A (Pirassununga/SP) foram fornecidas pelos voluntários (n=100) nos meses de Junho/2011, Setembro/2011, Dezembro/2011 e Marco/2012. Os voluntários do Grupo B (Erval Velho/SC) (n=20) forneceram amostras de alimentos no mês de Abril/2012. Em cada grupo, uma lista com 20 alimentos a base de milho foi entregue aos voluntários, para fornecimento de amostras daqueles disponíveis em suas respectivas residências em cada mês de amostragem, totalizando 122 amostras de derivados de milho no Grupo A e 17 amostras no Grupo B coletadas durante o estudo. Adicionalmente, aplicou-se um Questionário de Frequência Alimentar (QFA) e um Inquérito Recordatório de 24 horas (QIR - 24 h) no momento das coletas de amostras. Em cada mês de amostragem de alimentos, foram coletadas amostras de sangue, urina (somente Grupo A) e cabelo dos voluntários, sendo as amostras armazenadas a -20ºC (urina e cabelo) ou -80ºC (sangue) até o momento das análises. As amostras de alimentos foram submetidas a análise de FB1, sendo que as de farinha de milho foram também analisadas quanto ao teor de ácido fólico. Ambas as análises foram feitas através de cromatografia líquida de alta eficiência (CLAE). Em soro, foram avaliadas a relação esfinganina/esfingosina (Sa/So), resíduos de FB1, ácido fólico, uréia e creatinina. Em urina, foram analisados os níveis de FB1, creatinina para correção do volume urinário e a relação Sa/So. Em cabelo, foram analisados os resíduos de FB1 através de CLAE acoplada a espectrometria de massas. Todos os métodos de análise foram submetidos a procedimento de otimização e validação intra--laboratorial. A incidência de FB1 nos alimentos foi, em média, 72% (n=122) nas amostras do Grupo A (Pirassununga/SP) e 35% (n=17) no Grupo B (Erval Velho/SC). Os maiores níveis foram encontrados em amostras de pipoca provenientes do Grupo B, com uma amostra excedendo o limite de tolerância estabelecido no Brasil (2,500 &micro;g kg-1). A ingestão diária provável média (IDPM) de FB1 no Grupo A foi de 63,3 ng kg-1 peso corpóreo (p.c.) dia-1, que corresponde a 3,1% da ingestão provisória máxima tolerável (IPMT) recomendada para fumonisinas (2.000 ng kg-1 p.c. dia-1). A IDPM do Grupo B apresentou uma média de 190,1 ng kg-1 p.c. dia-1 o que corresponde a 9,5% da IDMT. As concentrações de ácido fólico nas amostras de farinha de milho variaram de < 0,3 &micro;g kg-1 (limite de quantificação do método) a 1.705 &micro;g kg-1, com média de 713 &plusmn; 435 &micro;g kg-1. Somente uma amostra apresentou nível de ácido fólico acima do valor mínimo estabelecido pela ANVISA. Em urina, a incidência de FB1 foi de 33,4% (n=251), com níveis médios de 3,19 &plusmn; 3,15 ng mg-1 de creatinina. Não houve correlação (P&gt;0,05) entre as concentrações de FB1 na urina e nos alimentos. Os níveis de esfinganina foram mais elevados em mulheres, com 25,0% (n=116) de amostras positivas, em comparação à urina de homens, 10,4% (n=96). A relação Sa/So apresentou em média 0,91, 0,77 e 0,89 para urina de mulheres, homens e em combinação, respectivamente. Em soro, os níveis de esfingosina foram em média 2,48 ng mL-1 para o Grupo A e 5,01 ng mL-1 para o Grupo B. A relação Sa/So variou de 0,06 a 3,19 com média de 0,79 para o Grupo A e 0,78 para o Grupo B. Embora tenha havido correlação positiva (r=0,574, P&lt;0,05) entre a relação Sa/So no soro e os dados de consumo de milho e derivados obtidos no QIR-24 h, não foram observadas correlações (P&gt;0,05) entre a ingestão de FB1 e a relação Sa/So na urina ou soro. A concentração de ácido fólico no soro variou de 6,7 a 24,0 ng mL-1 (média de 13,4 &plusmn; 5,4 ng mL-1), com ambos os grupos (A e B) apresentando resultados dentro dos valores de referências. Não foram observados níveis detectáveis de FB1 nas amostras de soro. No entanto, FB1 foi detectada em 4 amostras de cabelo humano (7,2%) dos Grupos A e B, cuja concentração média foi de 21,3 &plusmn; 12,1 ng g-1. Em síntese, os resultados obtidos nas análises de biomarcadores de FB1 no presente trabalho estão de acordo com os valores de IDPM encontrados, indicando que a exposição a FB1 nas populações estudadas não representa um risco a saúde. / Fumonisin B1 (FB1) is a mycotoxin produced by the secondary metabolism of Fusarium species, mainly F. verticillioides and F. proliferatum, which contaminates foods before and after processing and causes serious problems to public health and food quality. The aim of this study was to evaluate the human exposure to FB1 in food by means of estimated intake of toxin in the diet, and analysis of different biomarkers in serum, urine and hair. In addition, folic acid in food and blood as well urea and creatinin in serum were investigated to evaluate the toxin effects. The study was conducted in two cities of Sao Paulo and Santa Catarina States, where the respective volunteers were categorized as low-consumers of corn products (Group A, volunteers from Pirassununga/SP) and high-consumers of corn products (Group B, volunteers from Erval Velho/SC). Food samples from Group A (Pirassununga/SP) were provided by volunteers (n=100) in June/2011, September/2011, December/2011 and March/2012. The volunteers from Group B (Erval Velho/SC) (n=20) provided food samples in April/2012. In each group, a list of 20 corn products was given to volunteers, to allow them to check and collect the food items available in their homes at each sampling time. The total number of samples of corn products provided by the volunteers were 122 and 17 in Group A and Group B, respectively. Addicionally, a Food Frequency Questionnaire (FFQ) and a 24-Hours Dietary Recall Questionnaire (24h-DRQ) were applied by the time of sample collections. In each month of food samples collection, samples of blood, urine (only Group A) and hair from the volunteers were collected and storage at -20ºC (urine and hair) or -80ºC (blood) until analysis. Food samples were submitted to determination of FB1, and corn meal samples were also evaluated for folic acid levels. Both analysis were performed by high performance liquid chromatography (HPLC). In serum, analyses included sphinganine/sphingosine ratio (Sa/So), FB1 residue, folic acid, urea and creatinine. In urine, the levels of FB1, creatinine to correct urinary volume and Sa/So ratio were evaluated. In hair, FB1 residues were analysed by HPLC coupled to mass spectrometry. All the analytical methods were submitted to optimization and intra-laboratorial validation procedures. The mean incidences of FB1 in corn products were 72% (n=122) in samples of Group A (Pirassununga/SP), and 35% (n=17) of Group B (Erval Velho/SC). The higher levels were found in popcorn from Group B, with one sample exceeding the tolerance limit established in Brazil (2,500 &micro;g kg-1). The mean probable daily intake (PDIM) of FB1 in Group A was 63.3 ng kg-1 body weigh (b.w.) day-1, which corresponds to 3.1% of provisional maximum tolerable intake (PMTDI) recommended for fumonisins (2,000 ng kg-1 b.w. day-1). PDIM of Group B was 190.1 ng kg-1 b.w. day-1, which represents 9.5% of PMTDI. Folic acid levels in corn meal ranged from &LT; 0,3 &micro;g kg-1 (quantification limit) to 1.705 &micro;g kg-1, with a mean of 713 &plusmn; 435 &micro;g kg-1. Only one sample had levels of folic acid above the minimum established by ANVISA. In urine, the incidence of FB1 was 33,4% (n=251), at mean levels of 3,19 &plusmn; 3,15 ng mg-1 of creatinine. There wasn\'t correlation (P&gt;0.05) between concentrations of FB1 in urine and foods. Sphinganine levels were higher in woman, with 25.0% (n=116) of positive samples in comparison to urine of men, 10.4% (n=96). The mean Sa/So ratios were 0.91, 0.77 and 0.89 for urine of women, men and in combination, respectively. In serum, sphingosine presented a mean of 2.48 ng mL-1 to Group A and 5.01 ng mL-1 to Group B. Sa/So ratio ranged from 0.06 to 3.19 with a mean of 0.79 to Group A and 0.78 to Group B. Although a positive correlation (r=0.574, P&lt;0.05) was found between Sa/So ratio in serum and corn consumption data obtained by 24h-DRQ, no correlation was observed (P&gt;0,05) with FB1 intake and Sa/So ratio in urine or serum. Folic acid concentration in serum ranged from 6.7 to 24.0 ng mL-1 (mean of 13.4 &plusmn; 5.4 ng mL-1), with both groups (A and B) presenting levels within the reference valuies. There were no detectable levels of FB1 in serum samples. However, FB1 was detected in 4 human hair samples (7.2%) of Groups A and B, at a mean concentration was 21.3 &plusmn; 12.1 ng g-1. In summary, the results obtained in the analyses of FB1 biomarkers in the present study are in agreement with the PDIM values found, hence indicating that FB1 exposure in the populations studied do not represent a health concern.

abstract: Exposure to sun radiation (SUR) with ambient temperature may be an influencer on athletes’ sweat loss in different environments, but the results are not currently known. The purpose of this study was to determine the effects of SUR on fluid balance (FB) and hydration status (HS) in athletes exercising indoors and outdoors. Initial FB and HS were assessed in NCAA-DI female soccer athletes (n=10) of a single team in temperate, dry conditions (55-68°F, 18-48% humidity) who were monitored during 3 practices of equal estimated energy expenditure (EE): two outdoors in direct SUR (cold/moderate temperatures) and one indoors without SUR (moderate temperatures). Humidity, temperature, and wet bulb globe temperature (WBGT – a measurement partly based on SUR, including ambient temperature/relative humidity) were recorded using Heat Stress Meters placed in the direct sun or in the shade. Each athlete’s semi-nude dry body weight was recorded before and after exercise. Urine samples were taken before, after, and the morning after. Urine specific gravity (USG) was tested to assess HS. Athletes wore combined heart rate and activity monitors to estimate EE and were provided ad libitum water and/or a zero-calorie sports drink. Their total intake included weights of consumed food and drink. Sweat rate was calculated using body weight change and intakes of liquids minus urine losses/hour. Two-way repeated measures ANOVA analyzed group-level differences. No significance was found in total FB (1.01±0.32 L/hr) or EE/hr (444±97.1 kcal/hr) across all days (p>0.05). In analyzing individual athlete results, 40% had consistent USG >1.025 (p=0.001) suggesting potential dehydration. These 4 athletes selected water as their beverage, of which is known that consuming only water does not stimulate drinking behavior as does electrolyte drinks. The remaining 60% were overall not dehydrated (USG <1.025) but must be aware of incidental dehydration in hotter temperatures. The conclusion is that in low-moderate temperatures, athletes self-regulate drinking habits and achieve fluid balance during exercise with or without sun radiation. However, athletes with average USG >1.025 are likely to remain dehydrated in moderate temperatures. The findings suggest that more education would benefit these athletes by ensuring hydration in any environment. / Dissertation/Thesis / Masters Thesis Nutrition 2019

Objetivou-se avaliar o efeito da adição de sais de cálcio de ácidos graxos (SCAG) e de diferentes teores de proteína degradável no rúmen (PDR) na dieta de vacas leiteiras sobre: consumo, fermentação e metabolismo de nitrogênio ruminal, desempenho produtivo e balanço de nitrogênio; as estimativas de síntese microbiana utilizando 15N e bases purinas (BP) como indicadores microbianos, a composição dos pellets de bactéria, a recuperação de derivados purina (DP) na urina e a excreção urinária de DP de origem endógena; o fluxo omasal de ácidos graxos, a extensão de bio-hidrogenação e o perfil lipídico das bactérias. Oito vacas em lactação, multíparas e canuladas no rúmen foram usadas em um delineamento em quadrado Latino 4 × 4 replicado, distribuídas em arranjo fatorial 2 × 2 de tratamentos. Os tratamentos foram obtidos pela combinação da adição de SCAG (-SCAG: sem suplementação; e +SCAG: com 3,32% matéria seca (MS) de SCAG) e os teores de PDR (BPDR: baixo PDR, com 9,8% MS de PDR; e APDR: alto PDR: com 11,0% de PDR na MS da dieta). A suplementação lipídica reduziu (P &le; 0,049) o consumo, tendeu (P = 0,076) a aumentar o tempo de ruminação e aumentou (P = 0,015) o pH ruminal. A adição de SCAG diminuiu (P &le; 0,013) a digestibilidade aparente ruminal da MS e matéria orgânica, e tendeu a reduzir (P = 0,096) a digestibilidade ruminal da fibra em detergente neutro (FDN) e reduziu (P = 0,043) o nitrogênio (N) verdadeiramente digerido no rúmen. Apesar de afetar (P = 0,039) negativamente o fluxo de nitrogênio não amoniacal (NNA) associado com as grandes partículas, SCAG não afetou (P &ge; 0,634) o fluxo total de NNA e o fluxo de NNA microbiano, e aumentou (P = 0,032) a eficiência de síntese proteica microbiana. Animais alimentados com SCAG apresentaram maior (P = 0,043) produção de leite, com menor (P &lt; 0,001) teor de gordura, maior (P &lt; 0,001) eficiência alimentar e de uso do N. Animais alimentados com BPDR tenderam (P &le; 0,096) a apresentar maior consumo de proteína bruta e relação entre as concentrações ruminais de acetato e propionato. Ainda, APDR elevou (P &le; 0,048) a digestibilidade aparente total do amido e a digestibilidade ruminal verdadeira do N. O nível de PDR não afetou (P &ge; 0,719) a síntese de proteína microbiana e a eficiência de síntese, enquanto que BPDR aumentou (P &le; 0,055) a produção de leite e reduziu o teor de proteína do leite. O alto nível de PDR tendeu (P &le; 0,088) a reduzir a excreção fecal e o balanço de N, enquanto aumentou (P = 0,021) a excreção urinária relativa ao consumo de N. As bactérias associadas à partícula (BAP) apresentaram menor (P &le; 0,008) enriquecimento com 15N e relação N purinas: N total microbiano do que as bactérias associadas à fase líquida (BAL). A utilização de BP como indicador superestimou (P &le; 0,002) o fluxo microbiano, especialmente de BAL. No entanto, para o fluxo microbiano associado à partícula, houve tendência à interação (P = 0,067) entre os efeitos de indicador e da suplementação lipídica. Na presença de SCAG, BP e 15N apresentaram (P &gt; 0,05) estimativas similares, enquanto que na ausência de SCAG, BP subestimou (P &le; 0,05) o fluxo de BAP, em relação ao 15N. O aumento do nível de PDR da dieta tendeu a reduzir (P = 0,061) o fluxo omasal de BP e reduziu (P = 0,007) a recuperação urinária desta. A recuperação urinária de DP foi extremamente variável (33,9% de coeficiente de variação). As estratégias nutricionais avaliadas não afetaram (P &ge; 0,108) a excreção urinária de creatinina e DP de origem endógena. As excreções diárias de creatinina e DP de origem endógena foram de 0,262 mmol/kg e 640 mmol/kg0,75, respectivamente. O aumento do nível de PDR da dieta resultou na redução (P &lt; 0,001) do consumo dos ácidos graxos C4:0, C12:0, C14:0, C16:0 e C18:0, e tendeu (P = 0,068) a reduzir o consumo de C18:1 trans. Houve interação ou tendência (P &le; 0,095) à interação entre os efeitos do nível de PDR e adição de SCAG sobre o fluxo omasal de ácidos graxos totais e alguns ácidos graxos específicos (C15:0, C16:0, C17:0, C18:0, C18:1 cis-9, C18:2 cis-9 cis-12, C18:3 cis e C22:0). Nas dietas contendo SCAG, nenhum efeito do nível de PDR foi observado sobre o fluxo omasal de ácidos graxos (P &gt; 0,05), enquanto que nas dietas -SCAG, o aumento do nível de PDR aumentou (P &le; 0,05) o fluxo omasal total de ácidos graxos. Consequentemente, o aumento do nível de PDR reduziu (P &le; 0,05) a extensão de bio-hidrogenação do C18:1 cis e C18:3 nas dietas -SCAG, sem efeitos (P &gt; 0,05) nas dietas +SCAG. A adição de SCAG aumentou (P &le; 0,001) a extensão de bio-hidrogenação do C18:2, assim como o teor de AG dos pellets de bactéria. O nível de PDR da dieta não afetou (P &ge; 0,116) o perfil de ácidos graxos dos microrganismos, nem o perfil de ácidos graxos do leite. A adição de SCAG reduziu ou tendeu (P &le; 0,080) a reduzir os teores de C15:0, C16:0, C16:1, C17:0 e C18:0, e aumentou (P &le; 0,005) as concentrações de ácidos graxos insaturados de 18 carbonos nos microrganismos. Além disso, a adição de SCAG reduziu (P &le; 0,009) a concentração de todos os ácidos graxos menores do que 18 carbonos e a razão entre saturados e insaturados na gordura do leite, bem como aumentou (P &le; 0,001) as concentrações dos ácidos graxos com 18 carbonos ou mais e dos ácidos graxos insaturados. Desta forma, a adição de SCAG na dieta reduz o consumo, aumenta o tempo de ruminação, a eficiência microbiana e a produção de leite, sem afetar o fluxo microbiano. O aumento do nível de PDR da dieta reduz as perdas fecais de N e a produção de leite. As BP superestimam o fluxo microbiano, especialmente das BAL, pela presença de BP de origem dietética. A suplementação lipídica afeta a relação entre as estimativas de fluxo de BAP utilizando 15N e BP, levando à baixa correlação entre as estimativas com as duas técnicas. A recuperação urinária dos DP foi extremamente variável e aumentou com o aumento do nível de PDR da dieta. As excreções de creatinina e a excreção endógena de DP não são afetadas pela suplementação lipídica e pelo teor de PDR da dieta. A suplementação lipídica afeta consideravelmente o consumo e fluxo omasal de ácidos graxos, assim como o perfil lipídico dos microrganismos e o perfil de ácidos graxos do leite. Apesar de potencialmente aumentar o fluxo de alguns ácidos graxos insaturados e reduzir a extensão de bio-hidrogenação do C18:1 e C18:3, nas dietas sem SCAG, o nível de PDR tem poucos efeitos sobre o perfil de ácidos graxos das bactérias e do leite. / This study aimed to evaluate the effects of calcium salts of fatty acids (CSFA) addition and different levels of rumen degradable protein (RDP) in the diet of dairy cows on: feed intake, ruminal fermentation and nitrogen metabolism, productive performance and nitrogen balance; microbial protein synthesis estimates using 15N and purines basis (PB) as microbial markers, composition of bacterial pellets, urinary recovery of purines derivatives (PD) and endogenous PD excretion; omasal flow of fatty acids (FA), the bio-hydrogenation extent and FA profile of bacteria. Eight lactating, multiparous and rumen cannulated cows were used in a replicated 4 × 4 Latin square design, with a 2 × 2 factorial arrangement of treatments. The treatments were obtained by the combination of CSFA addition (-CSFA: without supplementation, and +CSFA: with 3.32% CSFA per kg dry matter - DM) and the RDP level (LRDP: low RDP, with 9.8% MS of RDP, and HRDP, high RDP: with 11.0% RDP of diet DM). Fat supplementation reduced (P &le; 0.049) feed intake, while tended (P = 0.076) to increase rumination time, and increased (P = 0.015) ruminal pH. In addition, CSFA supplementation decreased (P &le; 0.013) DM and organic matter ruminal digestibility, and tended (P &le; 0.096) to reduce NDF ruminal digestibility, and reduced (P = 0.043) N truly digested in the rumen. Besides negative effect (P = 0.039) on non-ammonia nitrogen (NAN) associated with large phase, CSFA showed no effect (P &ge; 0.634) on total NAN and microbial NAN flow, and increased (P = 0.032) microbial efficiency. Animals fed CSFA had higher (P = 0.043) milk yield, with lower (P &lt; 0.001) fat content, and improved (P &lt; 0.001) feed efficiency and N usage efficiency, in relation to those animals fed with non-supplemented diets. Animals fed LRDP tended to show higher (P &le; 0.096) CP intake, and acetate to propionate ruminal ratio, in relation to animals fed HRDP. In addition, HRDP increased (P &le; 0.048) starch total tract apparent digestibility and ruminal truly digestibility of N. The level of RDP showed no effect (P &ge; 0.719) on microbial N synthesis and microbial efficiency, and lower RDP level increased (P &le; 0.055) milk yield and decreased milk protein content. Decreased RDP level tended (P = 0.074) to reduce N fecal excretion, and increased (P = 0.021) fractional urinary excretion. Particle-associated bacteria (PAB) showed lower (P &le; 0.008) enrichment with 15N and purines N to total microbial N ratio, than liquid associated bacteria (LAB). Purine basis overestimated (P &le; 0.002) microbial flow, especially LAB flow. However, there was interaction (P = 0.067) between the effects of marker and lipid supplementation on PAB flow. In diets containing CSFA, PB and 15N showed similar (P > 0.05) estimates, whereas in the absence of CSFA, PB underestimated (P &le; 0.05) PAB flow, in relation to 15N. Increased dietary RDP levels tended (P = 0.061) to reduce omasal PB flow and decreased (P = 0.007) it\'s urinary recovery. Urinary recovery of PD was extremely variable (33.9% variation coefficient). Different evaluated factors had no effect (P &ge; 0.108) on urinary excretion of creatinine, and PD of endogenous origin. Daily excretions of creatinine and endogenous PD were 0.262 mmol/kg and 640 mmol/0.75, respectively. Higher RDP level resulted in reduced (P &lt; 0.001) C4:0, C12:0, C14:0, C16:0 and C18:0 FA intake, and tended (P = 0.068) to reduce C18:1 trans intake. There was an interaction (P &le; 0.095) between effects of the RDP and CSFA on the omasal flow of total FA and some specific FA (C15:0, C16:0, C17:0, C18:0, C18:1 cis-9, C18:2 cis-9 cis-12, C18: 3 cis and C22:0). This interaction occurred due to the higher (P &le; 0.05) omasal flow as consequence of increased RDP level, in the diets without CSFA. In diets containing CSFA, no effect (P &gt; 0.05) of the RDP level was observed on the omasal flow of FA. Consequently, the increase of RDP reduced (P &le; 0.05) the bio-hydrogenation extent of these FA in the diets -CSFA, with no effects (P &gt; 0.05) on the diets +CSFA. The addition of CSFA increased (P &le; 0.001) C18:2 bio-hydrogenation extent, as well as FA content of bacterial pellets. Dietary level of RDP had no effect (P &ge; 0.116) on microorganisms and milk FA profile. The addition of CSFA decreased or tended (P &le; 0.080) to decrease the levels of C15:0, C16:0, C16:1, C17:0 and C18:0 FA, while increased (P &le; 0.005) the concentrations of 18-carbon unsaturated fatty acids in the microorganisms. In addition, CSFA reduced (P &le; 0.009) the concentration of all FA lower than 18 carbons, saturated and unsaturated ratio in milk fat, and increased (P &le; 0.001) the concentrations of FA with 18 carbons or more, as well as unsaturated fatty acids. Thus, CSFA dietary addition decreases feed intake, increases rumination time, microbial efficiency, and milk yield. Higher level of RDP reduces N fecal losses and milk yield, and increases urinary losses of N. Purine basis overestimate microbial flow, especially LAB flow, by the presence of PB of dietary origin. Lipid supplementation affects the relationship between PAB flow estimates using 15N and PB, leading to a low correlation between the estimates with the two techniques. The urinary recovery of the PD was extremely variable and increased with the increase of the dietary RDP level. Creatinine excretions and endogenous excretion of PD are not affected by evaluated treatments. Lipid supplementation affects FA intake and omasal flow, as well as microorganisms and the milk FA profile. Although potentially increase the flow of some unsaturated FA and reduce the extent of bio-hydrogenation of C18:1 and C18:3 FA in the diets without CSFA, the level of RDP has no effects on microorganisms and milk FA profile.

Mycotoxins are secondary metabolites produced by fungi that occur naturally in foodstuffs, which can cause a large variety of toxic effects on vertebrates including humans. The objectives of this work were to evaluate the co-occurrence of 11 mycotoxins in food products, feed for broiler chicks, laying hens and dairy cattle, assess the human exposure to mycotoxins through food analysis versus consumption data and multimycotoxin biomarkers in urine, and characterize the associated risk of mycotoxin exposure in Brazilian rural areas. Sampling procedures were conducted in 38 small-scale dairy and poultry farms in the surroundings of Pirassununga and Descalvado (State of São Paulo), Pinhalzinho and Erval Velho (State of Santa Catarina). In these farms a total of 86 volunteers were recruited and instructed to provide samples of the morning urine (N = 162) in two sampling periods (April-May/2016 and December/2016), along with samples of rice (N = 66), bean (N = 59), wheat (N = 39), corn flour (N = 21) and corn meal (N = 18) available in their households. Samples of feed for broilers (N = 10), laying hens (N = 20) and dairy cattle (N = 15) were also collected. All samples were analyzed by ultra-performance liquid chromatography coupled to mass spectrometry (UPLC-MS/MS) for determination of aflatoxins (AF) B1, B2, G1 and G2, fumonisins (F) B1 and B2, ochratoxin A (OTA), zearalenone (ZEN), deoxynivalenol (DON), toxin T-2 and toxin HT-2 in food products and feeds, and AFM1, AFP1, AFQ1, FB1, OTA, T-2, HT-2, DON, de-epoxideoxynivalenol (DOM-1), ZEN, &alpha;-zearalenol (&alpha;-ZEL), &beta;-zearalenol (&beta;-ZEL) and 15-acetyl-DON in urine samples. The mycotoxin levels in urine were adjusted to creatinine concentration in each sample analyzed. In feed samples, median levels of total AF, total FB, ZEN and DON were 100 &micro;g/kg, 680 &micro;g/kg, 160 &micro;g/kg and 200 &micro;g/kg, respectively. The co-occurrence of two or more mycotoxins was confirmed in 51% of feed samples. Results indicate a high exposure of farm animals to mycotoxins in the feed, hence emphasizing the need to improve the feed quality regarding the contamination with mycotoxins in small-scale farms in Brazil, and the necessity of include feed in Brazilian regulation, especially for AF, FB, and ZEN. Mycotoxin levels above the Brazilian maximum permitted levels (MPL) were found in rice (1.5%), wheat flour (12.8%) and corn flour (14.3%) samples. Urine determinations revealed the presence of AFM1 and AFP1, DON, OTA, FB1 and ZEN at levels of 0.02-12.0 ng/mg creatinine. Regarding the probable daily intake (PDI) based on food data, only ZEN (0.156 &micro;g/kg b.w./day) had a Hazard Quotient (HQ) above the tolerance (&gt; 1). PDI values based on urinary levels for DON, OTA and total AF were 84.914, 0.031 and 0.001 &micro;g/kg b.w./day, respectively, resulting in HQ values &gt; 1, which may indicate health risks for the population studied. An informal intervention by means of educational activities and delivery of an information flyer during the first sampling period did not change the exposure levels to mycotoxins in the second sampling period. Further studies are needed to identify food items other than those analyzed in this work as sources of dietary mycotoxins, as well as the contribution of inhalation of contaminated dusts for the exposure. This is the first study to report the risk assessment of mycotoxins based on food and urinary levels in rural areas in Brazil. / As micotoxinas são metabólitos secundários produzidos por fungos que ocorrem naturalmente em alimentos, das quais podem causar uma grande variedade de efeitos tóxicos em vertebrados, incluindo humanos. Os objetivos deste trabalho foram avaliar a co-ocorrência de 11 micotoxinas em alimentos, rações para frangos de corte, poedeiras e gado leiteiro, avaliar a exposição humana a micotoxinas através de análise de alimentos versus dados de consumo e biomarcadores de múltiplas micotoxinas na urina, e caracterizar o risco associado de exposição a micotoxinas em áreas rurais brasileiras. Os procedimentos de amostragem foram conduzidos em 38 propriedades leiteiras e avícolas de pequeno porte nos arredores de Pirassununga e Descalvado (SP), Pinhalzinho e Erval Velho (SC). Nestas fazendas, um total de 86 voluntários foram recrutados e instruídos a fornecer amostras da primeira urina da manhã (N = 162) em dois períodos de amostragem (abril-maio/2016 e dezembro/2016), juntamente com amostras de arroz (N = 66) , feijão (N = 59), trigo (N = 39), farinha de milho (N = 21) e fubá (N = 18) disponíveis em suas residências. Amostras de ração para frangos de corte (N = 10), poedeiras (N = 20) e bovinos leiteiros (N = 15) também foram coletadas. Todas as amostras foram analisadas por cromatografia líquida de ultra-performance acoplada a espectrometria de massas (UPLC-MS/MS) para determinação de aflatoxinas (AF) B1, B2, G1 e G2, fumonisina (FB) B1 e B2, ocratoxina A (OTA), zearalenona (ZEN), desoxinivalenol (DON), toxina T-2 e toxina HT-2 em produtos alimentícios e rações, e AFM1, AFP1, AFQ1, FB1, OTA, T-2, HT-2, DON, de-epóxido-oxinivalenol (DOM-1), ZEN, &alpha;-zearalenol (&alpha;-ZEL), &beta;-zearalenol (&beta;-ZEL) e 15-acetil-DON em amostras de urina. Os níveis de micotoxinas na urina foram ajustados à concentração de creatinina em cada amostra analisada. Em amostras de ração, os níveis médios de AF total, FB total, ZEN e DON foram de 100 &micro;g/kg, 680 &micro;g/kg, 160 &micro;g/kg e 200 &micro;g/kg, respectivamente. A co-ocorrência de duas ou mais micotoxinas foi confirmada em 51% das amostras de ração. Os resultados indicam uma alta exposição de animais de fazenda à micotoxinas na ração, enfatizando a necessidade de melhorar a qualidade das rações em fazendas de pequena escala no Brasil, referente as micotoxinas, e a necessidade de incluir ração na legislação brasileira, especialmente para AF, FB e ZEN. Os níveis de micotoxinas acima dos níveis máximos permitidos no Brasil (LMP) foram encontrados em arroz (1,5%), farinha de trigo (12,8%) e farinha de milho (14,3%). As determinações da urina revelaram a presença de AFM1 e AFP1, DON, OTA, FB1 e ZEN nos níveis de 0,02-12,0 ng/mg de creatinina. Em relação à ingestão provável diária (IPD) com base em dados de alimentos, apenas ZEN (0,156 &micro;g/kg p.c./dia) apresentou um Quociente de Risco (HQ) acima do tolerável (&gt; 1). Os valores de IPD baseados nos níveis urinários para DON, OTA e AF total foram 84,914, 0,031 e 0,001 &micro;g/kg p.c./dia, respectivamente, resultando em valores de HQ&gt; 1, o que pode indicar riscos para a saúde da população estudada. Uma intervenção informal por meio de atividades educacionais e entrega de um folheto informativo durante o primeiro período de amostragem não alterou os níveis de exposição às micotoxinas no segundo período de amostragem. Mais estudos são necessários para identificar itens alimentares além dos analisados neste trabalho como fontes de micotoxinas diárias, bem como a contribuição da inalação de pós contaminados para a exposição. Este é o primeiro estudo a relatar a avaliação de risco de micotoxinas com base em alimentos e níveis urinários em áreas rurais no Brasil.

Urine comprises around 1% of domestic sewage volume but holds 80% of total nitrogen. Source separation is a sustainable way to wastewater management than traditional way due to low energy cost and preventing certain pollutants into wastewater treatment plants. Currently, removing and recovering nitrogen from source-separated urine has attracted more and more interests. Of them, ion exchange was used for removal and recovery of nitrogen in the form of ammonia from synthetic urine for potential application as a fertilizer in agriculture. No previous research studies were conducted to investigate the removal and recovery of nitrogen from hydrolyzed urine by ion exchange using POPs (porous organic polymers). So this study focused on evaluating the performance of POPs and comparing with clinoptilolite in synthetic hydrolyzed urine in terms of adsorption capacity (isotherm), adsorption rate (kinetics), regeneration rate, and cost. The ammonium removal from hydrolyzed urine using POPs was rapid with a high capacity of 68.03 mg/g than clinoptilolite (15.36 mg/g), and the regeneration efficiency of clinoptilolite and POPs can achieve 91% and 95.3%, respectively based single time use result. Although POPs had the better performance at one time use and multiple times use, it also had high materials cost. Additionally, the capacity of POP was estimated using the integrated ion exchange regeneration process model as 30.24 mg/g and 28.65 mg/g on cycle 10 and cycle 24, respectively. The regeneration efficiency of POPs was predicated as 45.4% and 38.4% in cycle 10 and cycle 24, respectively. The predicted capacity decreased with the number of cycles, but remained at about 55% of virgin POPs after 24 cycles, indicating POPs can maintain good performance after multiple reuses than clinoptilolite.

An analytical method was developed for the determination of the â-agonists clenbuterol, cimaterol and salbutamol in bovine retina. The method involved extraction into a pH 8.5 tris-HCl buffer, followed by protease enzyme digestion and immunoaffinity column cleanup before analysis by liquid chromatography with tandem mass spectrometry detection LC/MS/MS. The LOD for clenbuterol, salbutamol and cimaterol were 0.64, 1.20 and 1.92 ng/g respectively. The identities of the analytes were able to be confirmed to an acceptable standard. An analytical method was also developed for the analysis of the â-agonists clenbuterol, salbutamol, cimaterol, ractopamine, and mabuterol in bovine urine and emu muscle. The urine and muscle samples were digested with â-glcuronidase enzyme and cleaned up using a Bond Elute Certify SPE. The extracts were analysed by LC/MS/MS. Deuterated internal standards were used for quantitation. The LOD for urine [less than] 1ng g and for emu muscle it was [less than] 0.3 ng/g. The last part of the work describes the simultaneous gas chromatography-mass spectrometric analysis of diethylstilbestrol DES, hexestrol HEX, dienestrol DIEN, zeranol ZER, taleranol TAL and zeralenone ZON in fresh full cream and fresh skim milk. The analytes were analysed as their trimethyl silane (TMS) derivatives. A three phase solvent system was used for extraction and the extract was cleaned up using a combination of the anion exchange and hydrophobic properties of an anion exchange SPE. The detection limits for DES, DIEN, HEX, ZER, TAL and ZON were 9.6, 9.6, 16.8 , 7.2 , 13.5 and 34.8 ng/L respectively. / Doctor of Philosophy(PhD)

<p>Diverting urine from faeces or mixed wastewater and reusing it to fertilize crops, is a traditional method used in Asia. It is also a contemporary approach to sustainable nutrient and water management in Scandinavia and other parts of Europe. Urine diversion and reuse is a proven socio-technical system that has significant potential benefits on both a local and global scale, such as recirculating scarce plant nutrients like phosphorus back to agriculture, reducing eutrophication of waterways and improving water and sanitation systems. This thesis explores the nature of these benefits in Australia and the global context and what barriers would need to be overcome if a urine diversion and reuse system were implemented in Australia to achieve significant environmental benefits. These questions are investigated through stakeholder interviews in Sweden, to identify the ‘lessons learnt’ from the Swedish experience with urine diversion and reuse, and, through interviews with relevant stakeholders in Australia to identify possible barriers and opportunities, costs and benefits, and roles and responsibilities in the Australian context. Findings from both the stakeholder interviews are triangulated with other sources of knowledge, such as the literature, personal communications and a qualitative assessment of costs and benefits.</p><p>This thesis found that while urine diversion is likely to benefit the Australia situation and warrants further research, these benefits are fragmented and spread across a range of discourses and separate institutions. Its acceptance and effective introduction into Australia might therefore be challenged by its lack of a single obvious organisational home. To overcome this and other identified challenges, several recommendations are made. For example, an Australian demonstration trial of urine diversion and reuse is recommended where clear drivers and opportunities exist, such as: in new developments adjacent to agricultural land; in regions where algal blooms are a critical problem and are predominantly caused by municipal sewage discharges; and where synergies with waterless urinals are being considered for water conservation value. This thesis does not promote urine diversion and reuse as the ‘silver bullet’ to Australia’s water and nutrient problems, however it does recommend that it be considered on an equal basis next to other possible options. For example, if reducing nutrient loads on receiving water bodies is a key objective, then a cost-effective analysis of urine diversion and reuse, compared to other options to reduce nutrient loads, could be undertaken, ensuring all relevant costs and benefits to the whole of society are included in the analysis.</p>

<p>The Hull Street Integrated Housing Project, in Kimberley, is one of the projects supported by the Swedish International Development Cooperation Agency, Sida, in South Africa. The vision of the</p><p>project is to provide low cost housing for the people of Kimberley. As a way of ensuring sustainability, the project adopts the Ecological sanitation (Ecosan) approach where urine and</p><p>faeces are separated from the source.</p><p>The concept of Ecosan is new to many people around the world. To make the concept workable and acceptable effective implementation strategies are required.</p><p>At the Hull Street, after the first of the four phases 144 unit houses have been completed all fitted</p><p>with the UDS. Urine from the UDS as well as the greywater from the kitchen and bathroom are connected to infiltrate into the ground. This arrangement is called the “quick-fix”. The faeces from</p><p>the houses are sent to the compost yard for composting so that the residents could use the compost in their gardens.</p><p>This study which involves interview with some selected workers and residents in Hull Street</p><p>focuses on the modus operandi of the Ecosan unit of the Hull Street project with special emphasis on the methods of human excreta disposal and education strategies.</p>

<p>One of today’s most widely spread health problems is urinary incontinence, affecting 60-80% of the US population from age 15 and up. Treatment based on the possibility to implant a scaffold seeded with the patients’ own urine-derived stem cells, hUSC, to regenerate the damaged muscle tissue, would prove effective. A main challenge in regenerating new tissue from cell-seeded scaffolds is the limited cell survival due to insufficient oxygen diffusion to the center of the scaffold. Ways of enhancing cell survival, and thereby, proliferation and differentiation, is by hypoxic preconditioning of the cells or implantation in an oxygen-release material. Hypoxic preconditioning has shown to enhance proliferation as well as the expression of vascular endothelial growth factor, VEGF, in for example human bone marrow derived stem cells, hBMSC. VEGF is involved in the establishment of vasculature structures and an upregulation of its expression may therefore help promote quicker angeogenisis, increasing the oxygen supply and the cell survival. Oxygen-release materials have shown to enhance cell survival and growth both <em>in vitro</em> and <em>in vivo</em>.<em></em></p><p>This study aims to investigate the effect of hypoxia on hUSC, during 9 days of hypoxic culturing (2.0% ± 0.1% O₂) with and without oxygen-release material (PLGA 75:25 with 5 w% CPO) <em>in vitro</em>. hBMSC, and human smooth muscle cells, hSMC, have been used as control groups. Cell proliferation, morphology, differentiation, production of VEGF, and expression of hypoxia inducible factor HIF-1α have been studied.</p><p>According to the results, combining hypoxic preconditioning of hUSC with implantation in oxygen-release material could be an effective way to regenerate muscular tissue. Hypoxic preconditioning enhanced cell proliferation, production of VEGF, and HIF-1α expression. The increase of VEGF and HIF-1α would promote vascularization when implanted. The oxygen-release material showed possible promotion of cell differentiation, which would augment the hUSCs’ myogenic differentiation, while supplying oxygen until the tissue’s vascular structure has been established.</p>

The aims of the work this thesis is based upon were to develop convenient analytical procedures for determining resin acids in biological and environmental matrices, and apply them to enhance understanding of the occurrence, exposure to and uptake by exposed individuals of resin acids. Particular focus has been on the workplace environment of the Swedish wood pellets industry. Sample extraction procedures and high-performance liquid chromatography/electrospray ionisation-mass spectrometry (HPLC/ESI-MS) methodologies were developed for measuring resin acids in dust, skin and urine samples. Chromatographic separation of abietic (AA) and pimaric acid was achieved by using a polar-embedded C12 stationary phase. The HPLC/ESI-MS method avoids undesirable oxidation of AA, which was found to occur during the derivatisation step in the standard MDHS 83/2 gas chromatography/flame ionisation detection (GC/FID) methodology, leading to false observations of both AA and the oxidation product 7-oxodehydroabietic acid (7-OXO). Personal exposures to resin acids in the Swedish wood pellet production industry were found to be lower, on average, than the British Occupational Exposure Limit for rosin (50 µg/m3). The oxidised resin acid 7-OXO, was detected in both dust and skin samples indicating the presence of allergenic resin acids. A correlation between air and post-shift urinary concentrations of dehydroabietic acid (DHAA), and a trend towards an increase in urinary 7-OXO during work shifts, were also observed. Whether the increase in 7-OXO was due to direct uptake or metabolism of other resin acids cannot be concluded from the results. An efficient HPLC/UV methodology with diode-array detection was developed for screening commercial products for rosin that could be used in laboratories lacking mass spectrometers. Very high concentrations of free resin acids were detected in depilatory wax strips using the method. / At the time of doctoral defense, the following paper was unpublished and had a status as follows: Paper 4: Submitted.