<p>(From abstract) </p> <p>In the first study, the urine collection method was effectively applied for evaluation of intestinal permeability using Cr-EDTA, an indigestible oral marker, demonstrating the applicability of the procedure in 1-week-old and 6-week-old neonatal heifer calves (n=15 calves). Calf health observations were recorded during the entire urinary catheterization process and collection period to evaluate any negative health reactions to the procedure, or localized reactions. Proportion of localized reactions were analyzed, and the proportions did not exceed 20% for the calves catheterized at either 1 week or 6 weeks of age. </p> <p>In the second study, the developed catheterization procedure and urine collection method was applied using Cr-EDTA as an oral marker to investigate if L-GLN supplementation would offer improvement to intestinal permeability. In this larger study, 30 Holstein heifer calves [1.5 ± 0.5 days old; 37.1 ± 0.86 kg body weight (<strong>BW</strong>)] were blocked by serum total protein, BW, and age, and randomly assigned to 1 of 2 treatments: <strong>GLN</strong> [24% crude protein (<strong>CP</strong>)], 17% fat milk replacer (<strong>MR</strong>) +10 g L-GLN/kg MR powder) or <strong>NS</strong> (24% CP, 17% fat MR). MR was reconstituted to 12.5% solids with warm water and fed 3.8 L/calf/d until weaning. Calves were weaned at 56.4 ± 0.5 days of age, and had <em>ad libitum</em> grain (17% CP, 2% fat) and water access throughout the experimental period.</p> <p>During the preweaning period, calves were individually housed in hutches and health observations, which included respiratory and fecal scores, were assessed daily. Body weight was measured weekly, and grain and MR intake was assessed daily to calculate average daily gain (<strong>ADG</strong>), average daily feed intake [<strong>ADFI</strong>; grain intake (dry matter (<strong>DM)</strong> basis) + MR intake (DM basis)], and feed efficiency (<strong>G:F</strong>; ADG:ADFI). At weaning, calves were weighed, moved to pens (n = 3 pens/treatment, 4-5 calves/pen), provided free access to grain and grass hay, and then weighed 2 weeks post-weaning. Additionally, urinary catheters were placed at 1 and 6 weeks of age, and calves were orally dosed with 1 L Cr-EDTA in their MR. Urine samples were then collected over a 24-hr period for Cr output analysis as an <em>in vivo</em>biomarker of intestinal permeability. </p> <p>Blood was collected on study days 1, 2, 5, 7, 14, 21, 42, 56, and 70 to measure haptoglobin, serum amyloid A, leukocyte data, neutrophil: lymphocyte (<strong>N:L</strong>), glucose, non-esterified fatty acids, insulin, and cortisol. Two study periods were identified for data analysis representing greater (<strong>P1</strong>; weeks 1-3) and reduced (<strong>P2</strong>; weeks 4-8) enteric disease susceptibility. Data were analyzed using PROC GLIMMIX or PROC MIXED in SAS 9.4 with calf as the experimental unit. There was a decrease in total preweaning Cr output (<em>P</em> < 0.05) for GLN calves, and Cr output in 1 week old calves was decreased (<em>P</em> = 0.04) in GLN versus NS calves. The N:L was decreased overall (<em>P</em> = 0.03) and during P2 (<em>P</em> = 0.01) and P2 neutrophil count tended to be reduced (<em>P</em> = 0.07) in GLN versus NS calves. There were no MR treatment differences for ADFI, ADG, body measurements, post-absorptive metabolic biomarkers, disease scores, and therapeutic treatments (<em>P</em> > 0.10). In summary, L-GLN supplementation improved intestinal integrity and biomarkers of physiological stress in pre-weaned Holstein heifer calves managed under production-relevant conditions.  </p>

With reduced prevalence of visceral leishmaniasis (VL) in the Indian subcontinent (ISC), direct and field deployable diagnostic tests are needed to implement an effective diagnostic and surveillance algorithm for post-elimination VL control. In this regard, here we investigated the diagnostic efficacies of a loop-mediated isothermal amplification (LAMP) assay (Loopamp™ Leishmania Detection Kit, Eiken Chemical CO., Ltd, Japan), a real-time quantitative PCR assay (qPCR) and the Leishmania antigen ELISA (CLIN-TECH, UK) with different sampling techniques and evaluated their prospect to incorporate into post-elimination VL control strategies. Eighty clinically and rK39 rapid diagnostic test confirmed VL cases and 80 endemic healthy controls were enrolled in the study. Peripheral blood and dried blood spots (DBS) were collected from all the participants at the time of diagnosis. DNA was extracted from whole blood (WB) and DBS via silica columns (QIAGEN) and boil & spin (B&S) methods and tested with qPCR and Loopamp. Urine was collected from all participants at the time of diagnosis and was directly subjected to the Leishmania antigen ELISA. 41 patients were followed up and urine samples were collected at day 30 and day 180 after treatment and ELISA was performed. The sensitivities of the Loopamp-WB(B&S) and Loopamp-WB(QIA) were 96.2% (95% CI 89·43-99·22) and 95% (95% CI 87·69-98·62) respectively. The sensitivity of Loopamp- DBS(QIA) was 85% (95% CI 75·26- 92·00). The sensitivities of the qPCR-WB(QIA) and qPCR-DBS(QIA) were 93.8% (95% CI 86·01-97·94) and 72.5% (95% CI 61·38-81·90) respectively. The specificity of all molecular assays was 100%. The sensitivity and specificity of the Leishmania antigen ELISA were 97.5% (95% CI 91·47-99·70) and 91.95% (95% CI 84·12-96·70) respectively. The Leishmania antigen ELISA depicted clinical cure at day 180 in all the followed-up cases. Efficacy and sustainability identify the Loopamp-WB(B&S) and the Leishmania antigen ELISA as promising and minimally invasive VL diagnostic tools to support VL diagnostic and surveillance activities respectively in the post-elimination era.

No description available.

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Reindeer appear to have strong positive effects on plant productivity and nutrient cycling in strongly nutrient-limited ecosystems. While the direct effects of grazing on vegetation composition have been intensively studied, much less is known about the indirect effect of grazing on plant-soil interactions. This thesis investigated the indirect effects of ungulate grazing on arctic plant communities via soil nutrient availability and plant nutrient uptake. At high density, the deposition of dung alone increased plant productivity both in nutrient rich and nutrient poor tundra habitats without causing major changes in soil possesses. Plant community responses to dung addition was slow, with a delay of at least some years. By contrast, a 15N-urea tracer study revealed that nutrients from reindeer urine could be rapidly incorporated into arctic plant tissues. Soil and microbial N pools only sequestered small proportions of the tracer. This thesis therefore suggests a strong effect of dung and urine on plant productivity by directly providing nutrient-rich resources, rather than by stimulating soil microbial activities, N mineralization and ultimately increasing soil nutrient availability. Further, defoliation alone did not induce compensatory growth, but resulted in plants with higher nutrient contents. This grazing-induced increase in plant quality could drive the high N cycling in arctic secondary grasslands by providing litter of a better quality to the belowground system and thus increase organic matter decomposition and enhance soil nutrient availability. Finally, a 15N natural abundance study revealed that intense reindeer grazing influences how plants are taking up their nutrients and thus decreased plant N partitioning among coexisting plant species. Taken together these results demonstrate the central role of dung and urine and grazing-induced changes in plant quality for plant productivity. Soil nutrient concentrations alone do not reveal nutrient availability for plants since reindeer have a strong influence on how plants are taking up their nutrients. This thesis highlights that both direct and indirect effects of reindeer grazing are strong determinants of tundra ecosystem functioning. Therefore, their complex influence on the aboveground and belowground linkages should be integrated in future work on tundra ecosystem N dynamic.

This thesis describes studies on the analysis of arsenic (As) in human biological samples, mainly urine but also hair and fingernails using inductively coupled plasma mass spectrometry (ICP-MS) and graphite furnace atomic absorption spectrometry (GF-AAS). The relationship between ethnicity and arsenic metabolism was investigated for the first time for a population in the United Kingdom. This investigation has been carried out through comparative analysis of arsenic in human urine, hair and fingernails in volunteers from three different ethnic groups (Whites, Asians and Somali Black-Africans) who are only exposed to background levels of arsenic. Results obtained with 63 volunteers showed ethnic differences in urinary arsenic excretion as well as differences in arsenic levels in fingernail samples. The averages of total arsenic levels for the Somali Black-Africans (urine 7.2 µg/g creatinine; fingernails 723 µg/kg) are significantly (P< 0.05) different from both the Asians (urine 20.6 µg/g creatinine; fingernails 153.9 µg/kg) and Whites (urine 24.5 µg/g creatinine; fingernails 177.0 µg/kg). The Somali group also shows a higher percentage (50%) of dimethylarsinate (DMA) and a lower percentage (48%) of arsenobetaine (AB), compared to Asians (16% DMA and 83% AB) and Whites (22% DMA and 77% AB). The effect of fasting on urinary arsenic species distribution was also investigated by monitoring urine samples from 29 Ramadan fasting volunteers, with each volunteer providing a sample at the beginning (RF1) and at the end (RF2) of an approximately 12 hours fast. The results obtained showed the frequency of MA detection for RF2 was 12 and 2-fold higher than for the non-fasting and RF1 groups, respectively. This suggests fasting may alter the pattern of arsenic metabolism and excretion. However, there was no significant difference (P> 0.05) in the average of total level of arsenic for RF1 (18.3 µg/g creatinine) and RF2 (17.7 µg/g creatinine). A relationship between excretion of arsenic and selenium in individuals exposed to background levels of arsenic and selenium was investigated through analysis of urine samples from 93 volunteers from Leicester, UK. A positive correlation between arsenic and selenium was found and the As:Se ratio was 0.7 ± 0.4. The intra-individual variation of As:Se ratio does not alter significantly over time, as determined by monitoring urine samples from a volunteer over a period of one year. Furthermore, within a single day, with urine samples collected at the beginning and after a 12-hour fast, the As:Se ratio was found to be similar (0.7 ± 0.5). These findings suggest a close relationship between these two metalloids, the biological significance of which needs to be explored in the future.

Les pyréthrinoïdes et les pyréthrines sont des insecticides neurotoxiques auxquels on attribue également des effets néfastes sur les systèmes immunitaire, hormonal et reproducteur. Ils sont abondamment utilisés en agriculture, mais aussi en horticulture, en extermination et dans le traitement d’infestations parasitaires humaines et animales (gale, poux). Il y a donc un intérêt en santé environnementale à connaître l’ampleur de l’exposition humaine à ces pesticides. La mesure des métabolites urinaires des pyréthrinoïdes et des pyréthrines apparaît une approche de choix pour arriver à cette fin puisqu’elle permet, en théorie, d’obtenir un portrait global de l’exposition. Or,traditionnellement et par soucis de simplicité les concentrations volumiques ou ajustées pour la créatinine) de ces biomarqueurs dans des urines ponctuelles sont déterminées, mais l’effet de l’utilisation de ces unités sur la validité des estimations de dose quotidienne absorbée n’a jamais été vérifié. L’objectif général de cette thèse était donc de développer, appliquer et valider une nouvelle stratégie de mesure et d’analyse de biomarqueurs pour améliorer la précision et la fiabilité des évaluations de l’exposition individuelles et populationnelles aux pyréthrinoïdes et pyréthrines. Les objectifs spécifiques étaient : i) de caractériser l’exposition humaine à ces contaminants en région urbaine et rurale au Québec et ii) de comparer la validité de la nouvelle stratégie de mesure et d’analyse de biomarqueurs urinaires proposée avec les autres méthodes plus usuelles utilisées pour estimer l’exposition individuelle et populationnelle et les doses absorbées de pyréthrinoïdes. Des adultes et des enfants, recrutés dans la population de l’Île de Montréal et de la Montérégie ont recueilli leurs urines pendant une période d’au moins douze heures et complété un questionnaire documentant les sources potentielles d’exposition. Les quantités de métabolites de pyréthrinoïdes et pyréthrines (pmol) mesurées dans les urines ont été ajustées pour une période de douze heures exactement et pour le poids corporel. Les quantités excrétées en région urbaine ont été comparées à celles excrétées en région rurale et les données individuelles et populationnelles ont été comparées à celles qui auraient été obtenues si des concentrations volumiques ou ajustées pour la créatinine avaient été mesurées. Les résultats montrent que l’exposition à ces pesticides est ubiquiste puisque plus de 90% des participants excrétaient les principaux métabolites de pyréthrinoïdes et pyréthrines à un niveau supérieur au seuil de détection analytique. Les résultats suggèrent que l’alimentation pourrait être à l’origine de ce niveau de base puisque les autres sources d’exposition connues n’ont été que rarement rapportées. Au Québec, l’exposition en milieu rural apparaissait légèrement plus importante qu’en milieu urbain et certains facteurs d’exposition, comme l’utilisation de pesticides domestiques, ont été rapportés plus fréquemment en milieu rural. Enfin, il a été démontré que la mesure des concentrations volumiques ou ajustées pour la créatinine de biomarqueurs est une approche qui peut entraîner des biais importants (jusqu’à 500% d’erreur et plus) en particulier lors de l’évaluation de l’exposition individuelle. Il est évident que les autorités de santé publique et de santé environnementale employant des biomarqueurs urinaires afin d’évaluer l’exposition aux pyréthrinoïdes et aux pyréthrines (ainsi qu’à d’autres molécules ayant des demi-vies d’élimination similaire)devraient diriger leurs efforts vers la mesure des quantités excrétées pendant une période d’au moins douze heures pour obtenir un portrait le plus valide possible de l’exposition. Il serait aussi souhaitable de mieux documenter la toxicocinétique de ces molécules chez l’humain afin d’établir avec une plus grande confiance la relation entre les quantités excrétées de biomarqueurs et les doses absorbées de ces contaminants. / Pyrethroids and pyrethrins are neurotoxic insecticides also considered to have negative effects on the immune, endocrine and reproductive systems. They are abundantly used for agricultural and horticultural purposes, for pest control and to treat human and animal parasitic infestations (scabies, lice). Consequently, there is in environmental health an interest in evaluating the extent of human exposure to these pesticides. The measurement of pyrethroid and pyrethrin metabolites in urine seems to be the best approach because it provides in theory a global depiction of the exposure. Because of it straightforwardness, it is common practice to use the biomarkers volume-weighted or creatinine-adjusted concentrations in spot urine samples, however the validity of daily doses estimates derived from these units has yet to be assessed. The main goal of this research was to develop, apply and validate a new approach to the measurement and analysis of biomarkers to improve the precision and the reliability of estimates of pyrethroid and pyrethrin exposure at the individual and population levels. The specific objectives were: i) to characterize human exposure to these contaminants in urban and rural populations in Quebec and ii) to assess the validity of this new strategy of measurement and analysis of urinary biomarkers with the biological monitoring strategies generally used to assess individual and population pyrethroid exposure and absorbed doses. Adults and children recruited in the population of the Island of Montreal and of Monteregie collected their urines for at least twelve hours and filled a questionnaire about their potential sources of exposure. The amounts of pyrethroid and pyrethrin metabolites measured in urine (pmol) were adjusted to a fixed twelve hour period and for the body weight. The amounts excreted in the urban area were compared to those from the rural area and individual and population data were compared to those that would have been obtained if volume-weighted or creatinine-adjusted concentrations would have been used. Results show that exposure to these pesticides is very common, with more than 90% of the participants excreting the main pyrethroid and pyrethrin metabolites above the analytical limit of detection. These results also suggest that the diet could be the main contributor to this base level because the other known sources of exposure were rarely reported. In the province of Quebec, the exposure in a rural area seemed slightly more important than in an urban area and some exposure factors, like the use of household pesticides, were reported more frequently in rural area. Finally, it was shown that the measurement of volume-weighted or creatinine-adjusted concentrations is an approach that can lead to an important bias (an error of up to 500% and more) especially for the assessment of individual exposure. It becomes obvious that public health and environmental health authorities using urinary biomarkers to assess pyrethroid and pyrethrin exposure (or other compounds with similar half-lives) should focus their efforts on measuring the amounts excreted during a period of at least twelve hours to obtain the best picture of the exposure. It would also be pertinent to increase the knowledge of the toxicokinetic behaviour of these compounds in humans in order to establish with greater confidence the relation between the excreted amounts and the absorbed doses of these contaminants.

Urinvägsinfektion är en av de mest förekommande infektionerna hos människan. För att visualisera urinvägspatogener kan det kromogena mediet UriSelectTM4 användas för diagnostik. Det primära syftet med studien var att utvärdera om det är möjligt att inkubera det kromogena mediet UriSelectTM4 i 5 % koldioxid istället för aerob miljö utan att totalväxt och morfologi påverkas. Vidare utvärderades totalväxt och antal fria kolonier vid odling på en halv UriSelectTM4-agarplatta med två odlingstekniker för att undersöka om fortsatt diagnostik är möjligt. Urinprover inkuberades i aerob miljö och i 5 % koldioxid och jämfördes visuellt utifrån totalväxt, antal fria kolonier, morfologi samt färgförändring på kolonier och agarn. Resultatet visade att totalväxt och antal fria kolonier endast skiljer i liten grad mellan inkubationsmiljöerna. Däremot förekom skillnader i morfologi och färg. Vidare kunde en halv agarplatta användas vid odling och fortsatt diagnostik. Studien visar därmed att UriSelectTM4 kan inkuberas i 5 % koldioxid utan att totalväxt, fria kolonier och agarn påverkas. / Urinary tract infection is one of the most common infections among humans. For diagnostics, the chromogenic media UriSelectTM4 can be used to visualize the urinary tract pathogens. The primary purpose of the study was to evaluate if the chromogenic media UriSelectTM4 could be incubated in 5% carbon dioxide instead of aerobic environment without impacting total growth and morphology. Furthermore, total growth and number of free colonies was evaluated when cultivating on a half UriSelectTM4 agar media with two streak patterns to examine if further diagnostics is possible. Urine samples were incubated in aerobic environment and in 5% carbon dioxide and visually compared for total growth, number of free colonies, morphology and color change of bacterial colonies and the agar media. The results showed that total growth and free colonies only had slight differences between the incubation environments. On the other hand, morphology and color of the colonies may vary. Further a half agar media could be used for cultivation and further diagnostics. Consequently, the study shows that UriSelectTM4 can be incubated in 5% carbon dioxide without any impact on total growth, free colonies or of the chromogenic media.

Akut- och psykiatriska avdelningar på länssjukhuset Ryhov i Jönköping använder sig av snabbtest för drogscreening med varierande kvalitet under de tider då analysinstrumentet Konelab Prime 30i inte är bemannat. Syftet med studien var att utvärdera sensitivitet och specificitet hos Multitest 15 från tillverkaren Acro Biotech, och jämföra resultat från två olika avläsningstider. Antalet urinprover som samlades in för analys uppgick till 272. Positiva och negativa urinprover med drogkoncentrationer inom ±50% från varje drogs gränsvärde insamlades. Senare inkluderades drogkoncentrationer utanför detta intervall. Proverna testades med Multitest 15 vid laboratoriet för klinisk kemi på Ryhov efter utförd analys med Konelab Prime 30i, vars analysresultat utgjorde referens. De droger som testades var amfetamin, metamfetamin, ecstasy, bensodiazepiner, buprenorfin, kokain, metadon, morfin, THC, oxykodon och tramadol. För alla droger sammantaget var sensitiviteten 86,7% - 100%, specificiteten 33,3% - 100% och träffsäkerheten 71,4% - 94,7%. Provurvalet inom intervallet ±50% från gränsvärdet var begränsat, vilket avsevärt påverkat dessa beräkningar, och Konelab Prime 30i använder semikvantitativ metod vilken endast ger approximativa koncentrationsvärden som referens. / The emergency and psychiatric wards on the county hospital Ryhov in Jönköping utilize onsite drug testing with varying quality during evenings and night-time when no staff are operating the chemistry analyzer Konelab Prime 30i. The aim of the study is to evaluate the performance of sensitivity and specificity of Acro Biotech Multitest 15 and comparing results from two different reading-times. The number of urine samples collected for analysis was 272. Positive and negative urine samples with drug concentrations within ± 50% from cut-off were collected. Later, concentrations outside of this range was included. The samples were tested with Multitest 15 at the laboratory for clinical chemistry at Ryhov after analysis with Konelab Prime 30i providing reference results. The drugs tested were amphetamine, methamphetamine, ecstasy, benzodiazepines, buprenorphine, cocaine, methadone, morphine, THC, oxycodone and tramadol. All drugs included, the sensitivity was 86.7% - 100%, the specificity 33% - 100% and the accuracy 71.4% - 94.7%. The sample selection within the range ±50% from the cut-off value was limited, which significantly affected these calculations, and Konelab Prime 30i uses a semi-quantitative method only providing approximate concentration values for reference.

Nos dias atuais tem havido uma crescente preocupação com o consumo de álcool, bem como de outras drogas de abuso, no local de trabalho. Com isso, começaram a ser implantados programas que visam o controle e a prevenção do uso de álcool e drogas neste ambiente. De um modo geral, esses programas utilizam a urina dos trabalhadores para a realização de análises toxicológicas. O presente trabalho procurou validar a urina como amostra para determinação do etanol, através da (1) padronização de um método de análise e (2) estudo das correlações entre concentrações sanguínea, urinária e no ar exalado, obtidas de 10 voluntários saudáveis, após administração oral única de bebida alcoólica na dose de 0,68 g de etanol por kg de peso corpóreo, em coletas realizadas durante período de 7 horas. Foi utilizado 1 mL de amostra adicionada de n-propanol (padrão interno), separação por head space e cromatografia em fase gasosa com coluna Poraplot Q 25m x 0,32mm, tendo sido obtidos os seguintes resultados: tempo de retenção do etanol 2.718 ± 0,0024 min., limite de detecção em sangue e urina, 0,07 e 0,01g/l, respectivamente, precisão intra e interensaio igual a 11,4 e 10,0% para o sangue e 5,9 e 6,5% para a urina e recuperação relativa, 55,88 ± 8,03 a 146,35 ± 13,07% para o sangue e 91,60 ± 0,81 a 103,28 ± 1,79% para a urina. A comparação da técnica padronizada com a de imunofluorescência polarizada forneceu um r de 0,9976 para o sangue e 0,9949 para a urina. O estudo de conservação demonstrou que não houve produção in vitro de etanol nas amostras de urina, após permanência à temperatura ambiente por 1 semana. Os resultados de estabilidade mostraram que não houve variação estatisticamente significante entre os resultados de análises feitas em intervalo de 30 dias, quando armazenadas em freezer com adição de fluoreto de sódio a 1 % (p=0,1088 e 0,1548, para sangue e urina, respectivamente). A técnica padronizada foi utilizada nas amostras de sangue e urina colhidas dos voluntários, mostrando-se adequada para a detecção de etanol, em concentrações que variaram de 0,03 a 1,44 g/L. Os valores de correlação urina : sangue nos diferentes tempos de coleta mostraram menor variação que os de sangue: ar exalado, sendo que no período entre 2 e 5 horas após a ingestão do álcool houve os menores desvios de valores de correlação, em todos os indivíduos estudados. Assim sendo, a urina colhida no tempo de 3 horas após o início ou reinício da jornada de trabalho pode ser recomendada para análise de etanol, em programas de controle e prevenção do uso de álcool e drogas no local de trabalho. / Abstract not available.