Udder illnesses can have an essential influence on quantity and quality of the milk. A diagnosis is possible with the help of the deviation of milk contents, especially over the change of the somatic cells. The cell count consequently makes an important health and quality criterion of the milk. In order to determine the cell count and other contents directly after the vormilk, the relation between the count of the milk cells and the impedimetric quanities of the milk was examined systematically in a new measure cell with alternating current. The measurements were executed isotherm with different temperatures over a wide frequency area (10 Hz -13 MHz) and the results were compared with the cell counts obtained with standard procedures (Fossomatic). Relations between the measured complex electric quantities and the content of the milk like lactose and cell count were produced with help of statistical methods. No direct relation is found between the impedimetric qualities (complex condauctance, conductance or capacity) of milk and the number of cells found in it. Cause of that is the inhomogeneity of the cells, that varies considerably regarding their size and mass. The lactose concentration correlates good with impedimetric quantities (amplitude, phase shift, capacity and conductance) of milk. Particularly with the condactance of the milk a statistically secured relation is found over the entire frequency area (10 Hz - 13 MHz). The lactose concentration correlates on the other hand well with the cell count and makes a statistically secured prognosis of this quantity with high precision possible. Future it is possible to determine the cell count with the help of the complex conductivity, for what simple conductivity measuring instruments are suitable. / Eutererkrankungen können einen wesentlichen Einfluss auf Menge und Qualität der Milch haben. Eine Diagnose ist an Hand der Abweichung von Milchinhaltstoffen, insbesondere über die Veränderung der somatischen Zellen möglich. Die Zellzahl stellt somit ein wichtiges Gesundheits- und Qualitätskriterium der Milch. Um die Zellzahl und andere Inhaltsstoffe direkt nach dem ersten Gemelk zu bestimmen, wurde der Zusammenhang zwischen der Zahl der Milchzellen und den impedimetrischen Eigenschaften der Milch in neuartigen Messzelle mit Wechselstrom systematisch untersucht. Die Messungen wurden isotherm bei verschiedenen Temperaturen über einen weiten Frequenzbereich von 10 Hz - 13 MHz durchgeführt und die Ergebnisse mit denen nach Standardverfahren (Fossomatic) erhaltenen Zahlen verglichen. Mit Hilfe statistischer Methoden wurden Relationen zwischen den erhaltenen komplexen elektrischen Größen und den Inhaltstoffen der Milch wie Laktose und Zellzahl hergestellt. Es konnte kein direkter Zusammenhang zwischen den impedimetrischen Eigenschaften (komplexer Leitwert, Wirkleitwert oder Kapazität) der Milch und der Zahl darin befindlicher Zellen gefunden werden. Ursache dafür ist die Inhomogenität der Zellen, die hinsichtlich ihrer Größe und Masse beträchtlich variieren. Dagegen lässt sich die Laktosekonzentration gut mit den impedimetrischen Eigenschaften (Amplitude, Phasenverschiebung, Kapazität und Wirkleitwert) der Milch korrelieren. Besonders mit dem Wirkleitwert der Milch ist ein statistisch abgesicherter Zusammenhang über den gesamten Frequenzbereich (10 Hz bis 13 MHz) zu finden. Die Laktosekonzentration wiederum korreliert gut mit der Zellzahl und ermöglicht mit hoher Genauigkeit eine statistisch gesicherte Voraussage dieser Größe. Damit ist es möglich, zukünftig die Zellzahl über den Laktosegehalt anhand der komplexen Leitfähigkeit zu bestimmen, wozu einfache Leitfähigkeitsmessgeräte geeignet sind.

La mammite subclinique est un problème de santé fréquent et coûteux. Les infections intra-mammaires (IIM) sont souvent détectées à l’aide de mesures du comptage des cellules somatiques (CCS). La culture bactériologique du lait est cependant requise afin d’identifier le pathogène en cause. À cause de cette difficulté, pratiquement toutes les recherches sur la mammite subclinique ont été centrées sur la prévalence d’IIM et les facteurs de risque pour l’incidence ou l’élimination des IIM sont peu connus. L’objectif principal de cette thèse était d’identifier les facteurs de risque modifiables associés à l’incidence, l’élimination et la prévalence d’IIM d’importance dans les troupeaux laitiers Canadiens. En premier lieu, une revue systématique de la littérature sur les associations entre pratiques utilisées à la ferme et CCS a été réalisée. Les pratiques de gestion constamment associées au CCS ont été identifiées et différentiées de celles faisant l’objet de rapports anecdotiques. Par la suite, un questionnaire bilingue a été développé, validé, et utilisé afin de mesurer les pratiques de gestion d’un échantillon de 90 troupeaux laitiers canadiens. Afin de valider l’outil, des mesures de répétabilité et de validité des items composant le questionnaire ont été analysées et une évaluation de l’équivalence des versions anglaise et française a été réalisée. Ces analyses ont permis d’identifier des items problématiques qui ont du être recatégorisés, lorsque possible, ou exclus des analyses subséquentes pour assurer une certaine qualité des données. La plupart des troupeaux étudiés utilisaient déjà la désinfection post-traite des trayons et le traitement universel des vaches au tarissement, mais beaucoup des pratiques recommandées n’étaient que peu utilisées. Ensuite, les facteurs de risque modifiables associés à l’incidence, à l’élimination et à la prévalence d’IIM à Staphylococcus aureus ont été investigués de manière longitudinale sur les 90 troupeaux sélectionnés. L’incidence d’IIM semblait être un déterminant plus important de la prévalence d’IIM du troupeau comparativement à l’élimination des IIM. Le port de gants durant la traite, la désinfection pré-traite des trayons, de même qu’une condition adéquate des bouts de trayons démontraient des associations désirables avec les différentes mesures d’IIM. Ces résultats viennent souligner l’importance des procédures de traite pour l’obtention d’une réduction à long-terme de la prévalence d’IIM. Finalement, les facteurs de risque modifiables associés à l’incidence, à l’élimination et à la prévalence d’IIM à staphylocoques coagulase-négatif (SCN) ont été étudiés de manière similaire. Cependant, afin de prendre en considération les limitations de la culture bactériologique du lait pour l’identification des IIM causées par ce groupe de pathogènes, une approche semi-Bayesienne à l’aide de modèles de variable à classe latente a été utilisée. Les estimés non-ajusté de l’incidence, de l’élimination, de la prévalence et des associations avec les expositions apparaissaient tous considérablement biaisés par les imperfections de la procédure diagnostique. Ce biais était en général vers la valeur nulle. Encore une fois, l’incidence d’IIM était le principal déterminant de la prévalence d’IIM des troupeaux. Les litières de sable et de produits du bois, de même que l’accès au pâturage étaient associés à une incidence et une prévalence plus basse de SCN. / Subclinical mastitis is a very frequent and costly health issue that can be detected using somatic cell count (SCC) measurements, but requires the use of milk bacteriological culture for identification of the causal pathogen. Because of this latter difficulty, nearly all subclinical mastitis research has focused on prevalent intramammary infections (IMI) and less is known on risk factors for IMI incidence or elimination. The main objective of this thesis was to identify the manageable risk factors associated with the incidence, elimination, and prevalence of IMI of importance in Canadian dairy herds. First, a systematic review of the literature on the associations between management practices used on farms and SCC was carried out. Management practices consistently associated with SCC were identified and differentiated from other practices for which anecdotic reports were available. Then, a bilingual questionnaire was developed, validated, and employed to measure the practices used on a sample of 90 Canadian dairy herds. To validate this tool, measures of repeatability and of validity of the questionnaire’s items were analyzed and an evaluation of the equivalence of the English and French versions was conducted. These analyses indicated that the questionnaire was, in general, acceptable, but also pinpointed some problematic items. These items were recategorized when possible or otherwise excluded from subsequent analyses to ensure good data quality. Most of the herds studied were already using post-milking teat disinfection and blanket dry cow therapy. Many other frequently recommended practices were not widely adopted. Next, manageable risk factors associated with Staphylococcus aureus IMI incidence, elimination, and prevalence were investigated on the 90 selected herds in a longitudinal fashion. The S. aureus IMI incidence appeared to be a stronger determinant of the herd prevalence than the elimination rate. Among other practices, wearing gloves during milking, using pre-milking teat disinfection, and having an adequate teat end condition showed desirable associations with the outcomes. These results highlight the importance of good milking practices to achieve a long-term reduction of S. aureus IMI prevalence. Finally, manageable risk factors associated with coagulase-negative staphylococci (CNS) IMI incidence, elimination, and prevalence were investigated in a similar manner. Because of the greater limitations of milk bacteriological culture to identify IMI caused by this group of pathogens, a semi-Bayesian latent class model approach was used. Sensitivity and specificity estimates from an internal validation study were used to link the observed IMI milk culture result to the latent true quarter IMI status. Non-adjusted estimates of IMI incidence, elimination, prevalence, and of associations with expositions appeared to be considerably biased by the diagnostic procedure’s imperfections. Most often, estimates were biased toward the null value. Again, IMI incidence was the main determinant of the herd IMI prevalence. Sand and wood products bedding were associated with lower CNS incidence and prevalence. Sending cows to pasture was also associated with lower CNS IMI incidence and prevalence.

INTRODUÇÃO: Estudos experimentais suportam a evidência de leucopenia persistente desencadeada pela morte encefálica (ME). OBJETIVO: Esse estudo teve como objetivo investigar o comportamento leucocitário na medula óssea e no sangue após a morte encefálica em ratos. MÉTODOS: A morte encefálica foi induzida através da inserção e insuflação rápida de um cateter no espaço intracraniano. Ratos falso-operados (FO) foram apenas trepanados. Decorridas seis horas, as células da medula óssea, coletadas da cavidade femural, foram utilizadas para as contagens total e diferencial e analisadas por citometria de fluxo para a caracterização das subpopulações linfocitárias, a expressão de moléculas de adesão granulocíticas e apoptose/necrose (método de Anexina V/Iodeto de Propídio (PI)). RESULTADOS: Ratos com ME apresentaram uma redução de 30% no número de células da medula óssea devido à redução de linfócitos (40%) e células segmentadas (45%). As subpopulações de linfócitos na medula óssea foram semelhantes nos animais ME e FO (CD3, p=0,1; CD4, p=0,4; CD3/CD4, p=0,4; CD5, p=0,4, CD3/CD5, p=0,2; CD8, p=0,8). A expressão de L-selectina e beta2-Integrinas nos granulócitos também não diferiram entre os grupos (CD11a, p=0,9; CD11b/c, p=0,7; CD62L, p=0,1). Não existem diferenças nas porcentagens de apoptose e de necrose (Anexina V, p=0,73; PI, p=0,21; Anexina V/PI, p=0,29). CONCLUSÃO: Os dados sugerem que a redução na mobilização de células da medula óssea para o sangue, desencadeada pela morte encefálica, não se relaciona a alterações de subpopulações de linfócitos, expressão de moléculas de adesão granulocíticas, ou apoptose e necrose / INTRODUCTION: Experimental findings support the evidence of a persistent leucopenia triggered by brain death (BD). AIMS: This study aimed to investigate leukocyte behavior in bone marrow and blood after BD in rats. METHODS: BD was induced by quickly inflation of an intracranial balloon catheter. Sham operated (SH) rats were trepanned only. Six hours thereafter bone marrow cells harvested from the femoral cavity were used for total and differential counts, and analyzed by flow cytometry to characterize lymphocyte subsets, granulocyte adhesion molecules expression, and apoptosis/necrosis (annexin V/propidium iodide (PI) protocol). RESULTS: BD rats exhibited a 30% reduction in bone marrow cells due to a reduction in lymphocytes (40%) and segmented cells (45%). Bone marrow lymphocyte subsets were similar in BD and SH rats (CD3, p=0.1; CD4, p=0.4; CD3/CD4, p=0.4; CD5, p=0.4, CD3/CD5, p=0.2; CD8, p=0.8). Expression of L-selectin and ?2-integrins on granulocytes did not differ (CD11a, p=0.9; CD11b/c, p=0.7; CD62L, p=0.1). There were no differences in the percentage of apoptosis and necrosis (Annexin V, p=0.73; PI, p=0.21; Annexin V/PI, p=0.29). CONCLUSIONS: Data presented suggest that the down-regulation of the bone marrow triggered by BD is not related to changes in lymphocyte subsets, granulocyte adhesion molecules expression, or apoptosis and necrosis

In der vorliegenden Arbeit wurde die Bedeutung des Einflusses von Laktationsnummer, Laktationsstadium, Körperkondition und Absetzverfahren auf Merkmale der Milchleistung und Eutergesundheit einer Herde Ostfriesischer Milchschafe untersucht. Die Tiere wurden nach dem Ablammen zufällig einer Früh- bzw. Spätabsetzergruppe zugeordnet (Absetzen der Lämmer 3 bzw. 42 Tage post partum). An insgesamt 40 Terminen wurden über eine vollständige Laktation Vorgemelks-, Hälftenanfangsgemelks- und Einzeltiergemelksproben gewonnen. Das Spektrum der untersuchten Merkmale umfasste die Milchmenge, die Milchinhaltsstoffe (Fett, Eiweiß, Laktose), die klinische Untersuchung des Euters, die somatische Zellzahl, die elektrische Leitfähigkeit und bakteriologische Untersuchungen. Zusätzlich wurden das Körpergewicht und die Körperkondition der Mutterschafe erfasst. Das durchschnittliche Leistungsniveau der Herde betrug 301±101,3 kg Milch, bei mittleren Milchfett-, Milcheiweiß-, und Laktosegehalten von 5,00, 5,14 bzw. 5,00 % (150-Tageleistung). Die Laktationsnummer hatte keinen signifikanten Effekt auf die Milchleistung. Schafe der Spätabsetzer-Gruppe produzierten im Anschluss an die Säugephase eine um 300 g signifikant höhere Testtagsmilchmenge als Tiere der Frühabsetzer-Gruppe. Das Absetzverfahren hatte keinen nachweisbaren Effekt auf die Milchinhaltsstoffe. Die Herde wies eine gute Eutergesundheit auf. Der Anteil bakteriologisch positiver Schafmilchproben betrug 28,5 %. Als dominante Erreger wurden in 96,6 % der bakteriologisch positiven Schafmilchproben Koagulasenegative Staphylokokken nachgewiesen. Der Anteil bakteriologisch positiver Euterhälftenbefunde stieg signifikant mit dem Fortschreiten der Laktation und zunehmender Laktationsnummer. Eine Euterinfektion mit Koagulasenegativen Staphylokokken sowie das Verfahren des Spätabsetzens beeinflussten alle drei untersuchten Eutergesundheitsparameter (logarithmierte Zellzahl, elektrische Leitfähigkeit und Laktosegehalt) negativ (p < 0,001).

La mammite subclinique est un problème de santé fréquent et coûteux. Les infections intra-mammaires (IIM) sont souvent détectées à l’aide de mesures du comptage des cellules somatiques (CCS). La culture bactériologique du lait est cependant requise afin d’identifier le pathogène en cause. À cause de cette difficulté, pratiquement toutes les recherches sur la mammite subclinique ont été centrées sur la prévalence d’IIM et les facteurs de risque pour l’incidence ou l’élimination des IIM sont peu connus. L’objectif principal de cette thèse était d’identifier les facteurs de risque modifiables associés à l’incidence, l’élimination et la prévalence d’IIM d’importance dans les troupeaux laitiers Canadiens. En premier lieu, une revue systématique de la littérature sur les associations entre pratiques utilisées à la ferme et CCS a été réalisée. Les pratiques de gestion constamment associées au CCS ont été identifiées et différentiées de celles faisant l’objet de rapports anecdotiques. Par la suite, un questionnaire bilingue a été développé, validé, et utilisé afin de mesurer les pratiques de gestion d’un échantillon de 90 troupeaux laitiers canadiens. Afin de valider l’outil, des mesures de répétabilité et de validité des items composant le questionnaire ont été analysées et une évaluation de l’équivalence des versions anglaise et française a été réalisée. Ces analyses ont permis d’identifier des items problématiques qui ont du être recatégorisés, lorsque possible, ou exclus des analyses subséquentes pour assurer une certaine qualité des données. La plupart des troupeaux étudiés utilisaient déjà la désinfection post-traite des trayons et le traitement universel des vaches au tarissement, mais beaucoup des pratiques recommandées n’étaient que peu utilisées. Ensuite, les facteurs de risque modifiables associés à l’incidence, à l’élimination et à la prévalence d’IIM à Staphylococcus aureus ont été investigués de manière longitudinale sur les 90 troupeaux sélectionnés. L’incidence d’IIM semblait être un déterminant plus important de la prévalence d’IIM du troupeau comparativement à l’élimination des IIM. Le port de gants durant la traite, la désinfection pré-traite des trayons, de même qu’une condition adéquate des bouts de trayons démontraient des associations désirables avec les différentes mesures d’IIM. Ces résultats viennent souligner l’importance des procédures de traite pour l’obtention d’une réduction à long-terme de la prévalence d’IIM. Finalement, les facteurs de risque modifiables associés à l’incidence, à l’élimination et à la prévalence d’IIM à staphylocoques coagulase-négatif (SCN) ont été étudiés de manière similaire. Cependant, afin de prendre en considération les limitations de la culture bactériologique du lait pour l’identification des IIM causées par ce groupe de pathogènes, une approche semi-Bayesienne à l’aide de modèles de variable à classe latente a été utilisée. Les estimés non-ajusté de l’incidence, de l’élimination, de la prévalence et des associations avec les expositions apparaissaient tous considérablement biaisés par les imperfections de la procédure diagnostique. Ce biais était en général vers la valeur nulle. Encore une fois, l’incidence d’IIM était le principal déterminant de la prévalence d’IIM des troupeaux. Les litières de sable et de produits du bois, de même que l’accès au pâturage étaient associés à une incidence et une prévalence plus basse de SCN. / Subclinical mastitis is a very frequent and costly health issue that can be detected using somatic cell count (SCC) measurements, but requires the use of milk bacteriological culture for identification of the causal pathogen. Because of this latter difficulty, nearly all subclinical mastitis research has focused on prevalent intramammary infections (IMI) and less is known on risk factors for IMI incidence or elimination. The main objective of this thesis was to identify the manageable risk factors associated with the incidence, elimination, and prevalence of IMI of importance in Canadian dairy herds. First, a systematic review of the literature on the associations between management practices used on farms and SCC was carried out. Management practices consistently associated with SCC were identified and differentiated from other practices for which anecdotic reports were available. Then, a bilingual questionnaire was developed, validated, and employed to measure the practices used on a sample of 90 Canadian dairy herds. To validate this tool, measures of repeatability and of validity of the questionnaire’s items were analyzed and an evaluation of the equivalence of the English and French versions was conducted. These analyses indicated that the questionnaire was, in general, acceptable, but also pinpointed some problematic items. These items were recategorized when possible or otherwise excluded from subsequent analyses to ensure good data quality. Most of the herds studied were already using post-milking teat disinfection and blanket dry cow therapy. Many other frequently recommended practices were not widely adopted. Next, manageable risk factors associated with Staphylococcus aureus IMI incidence, elimination, and prevalence were investigated on the 90 selected herds in a longitudinal fashion. The S. aureus IMI incidence appeared to be a stronger determinant of the herd prevalence than the elimination rate. Among other practices, wearing gloves during milking, using pre-milking teat disinfection, and having an adequate teat end condition showed desirable associations with the outcomes. These results highlight the importance of good milking practices to achieve a long-term reduction of S. aureus IMI prevalence. Finally, manageable risk factors associated with coagulase-negative staphylococci (CNS) IMI incidence, elimination, and prevalence were investigated in a similar manner. Because of the greater limitations of milk bacteriological culture to identify IMI caused by this group of pathogens, a semi-Bayesian latent class model approach was used. Sensitivity and specificity estimates from an internal validation study were used to link the observed IMI milk culture result to the latent true quarter IMI status. Non-adjusted estimates of IMI incidence, elimination, prevalence, and of associations with expositions appeared to be considerably biased by the diagnostic procedure’s imperfections. Most often, estimates were biased toward the null value. Again, IMI incidence was the main determinant of the herd IMI prevalence. Sand and wood products bedding were associated with lower CNS incidence and prevalence. Sending cows to pasture was also associated with lower CNS IMI incidence and prevalence.

Animal traits differ not only in mean, but also in variation around the mean. For instance, one sire’s daughter group may be very homogeneous, while another sire’s daughters are much more heterogeneous in performance. The difference in residual variance can partially be explained by genetic differences. Models for such genetic heterogeneity of environmental variance include genetic effects for the mean and residual variance, and a correlation between the genetic effects for the mean and residual variance to measure how the residual variance might vary with the mean. The aim of this thesis was to develop a method based on double hierarchical generalized linear models for estimating genetic heteroscedasticity, and to apply it on four traits in two domestic animal species; teat count and litter size in pigs, and milk production and somatic cell count in dairy cows. The method developed is fast and has been implemented in software that is widely used in animal breeding, which makes it convenient to use. It is based on an approximation of double hierarchical generalized linear models by normal distributions. When having repeated observations on individuals or genetic groups, the estimates were found to be unbiased. For the traits studied, the estimated heritability values for the mean and the residual variance, and the genetic coefficients of variation, were found in the usual ranges reported. The genetic correlation between mean and residual variance was estimated for the pig traits only, and was found to be favorable for litter size, but unfavorable for teat count.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The study aims to evaluate the effect of somatic cell count (SCC) and total bacterial count (TBC) on the constituents of milk, checking the influence that the climatic variables have on these hygienic-sanitary indicators. Data were obtained from 1,541 dairy farms located in 15 municipalities in the dairy region of Vale do Taquari, Rio Grande do Sul. It was tabulated the data from somatic cell count (SCC), total bacterial count (TBC) and milk composition, from June 2008 to December 2011, totaling 44,089 samples. The ambient temperature showed positive and significant correlation to score of somatic cell, while rainfall and relative humidity air showed no correlation. Moreover, the climatic variables have no significant correlation to the TBC. The fat, protein, minerals and total solids were directly correlated with the SCC and TBC, while solids-not-fat and lactose showed an opposite behavior. The principal component analysis (PCA) followed by agglomerative hierarchical clustering method, have showed that milk from SCC 401,000 to 750,000 cell mL-1 and TBC from 100,000 to 1,000,000 CFU mL-1, have the same quality, not justifying stratification in these intervals. In addition, it was observed that only limits regulatory standards are not sufficient to improve the quality of milk, suggesting parameters for payment system based on the bonus and penalty relative to the TBC and SCC milk. / Este estudo teve por objetivos avaliar o efeito da contagem de células somáticas (CCS) e da contagem bacteriana total (CBT) sobre os constituintes do leite, verificando a influência que as variáveis climáticas exercem sobre estes indicadores higiênico-sanitários. Os dados utilizados foram de 1.541 unidades produtoras de leite referentes a 15 municípios da bacia leiteira do Vale do Taquari, Rio Grande do Sul. Foram tabulados os dados de contagem de células somáticas (CCS), contagem bacteriana total (CBT) e composição centesimal do leite, referentes ao período de junho de 2008 a dezembro de 2011, totalizando 44.089 amostras analisadas. A temperatura ambiente apresentou correlação positiva e significativa com o escore de células somáticas, enquanto a precipitação pluviométrica e umidade relativa do ar ausência de correlação. Por outro lado, as variáveis climáticas não apresentaram correlação significativa com a CBT. Os teores de gordura, proteína, minerais e sólidos totais foram diretamente correlacionados com a CCS e a CBT, enquanto que os sólidos não gordurosos e a lactose apresentaram comportamento inverso. A análise de componentes principais (ACP) seguida pelo método hierárquico aglomerativo de agrupamento, permitiu constatar que leites com CCS maiores que 400.000 até 750.000 céls mL-1 e CBT superiores a 100.000 até 1.000.000 UFC mL-1, apresentam a mesma qualidade, não justificando a estratificação de intervalos nesta amplitude de variação. Paralelamente, observou-se que somente limites de normas regulatórias não são suficientes para melhoraria da qualidade do leite, sugerindo-se, parâmetros para sistemas de pagamento baseado na bonificação e penalização em relação à CCS e CBT do leite.

INTRODUÇÃO: Estudos experimentais suportam a evidência de leucopenia persistente desencadeada pela morte encefálica (ME). OBJETIVO: Esse estudo teve como objetivo investigar o comportamento leucocitário na medula óssea e no sangue após a morte encefálica em ratos. MÉTODOS: A morte encefálica foi induzida através da inserção e insuflação rápida de um cateter no espaço intracraniano. Ratos falso-operados (FO) foram apenas trepanados. Decorridas seis horas, as células da medula óssea, coletadas da cavidade femural, foram utilizadas para as contagens total e diferencial e analisadas por citometria de fluxo para a caracterização das subpopulações linfocitárias, a expressão de moléculas de adesão granulocíticas e apoptose/necrose (método de Anexina V/Iodeto de Propídio (PI)). RESULTADOS: Ratos com ME apresentaram uma redução de 30% no número de células da medula óssea devido à redução de linfócitos (40%) e células segmentadas (45%). As subpopulações de linfócitos na medula óssea foram semelhantes nos animais ME e FO (CD3, p=0,1; CD4, p=0,4; CD3/CD4, p=0,4; CD5, p=0,4, CD3/CD5, p=0,2; CD8, p=0,8). A expressão de L-selectina e beta2-Integrinas nos granulócitos também não diferiram entre os grupos (CD11a, p=0,9; CD11b/c, p=0,7; CD62L, p=0,1). Não existem diferenças nas porcentagens de apoptose e de necrose (Anexina V, p=0,73; PI, p=0,21; Anexina V/PI, p=0,29). CONCLUSÃO: Os dados sugerem que a redução na mobilização de células da medula óssea para o sangue, desencadeada pela morte encefálica, não se relaciona a alterações de subpopulações de linfócitos, expressão de moléculas de adesão granulocíticas, ou apoptose e necrose / INTRODUCTION: Experimental findings support the evidence of a persistent leucopenia triggered by brain death (BD). AIMS: This study aimed to investigate leukocyte behavior in bone marrow and blood after BD in rats. METHODS: BD was induced by quickly inflation of an intracranial balloon catheter. Sham operated (SH) rats were trepanned only. Six hours thereafter bone marrow cells harvested from the femoral cavity were used for total and differential counts, and analyzed by flow cytometry to characterize lymphocyte subsets, granulocyte adhesion molecules expression, and apoptosis/necrosis (annexin V/propidium iodide (PI) protocol). RESULTS: BD rats exhibited a 30% reduction in bone marrow cells due to a reduction in lymphocytes (40%) and segmented cells (45%). Bone marrow lymphocyte subsets were similar in BD and SH rats (CD3, p=0.1; CD4, p=0.4; CD3/CD4, p=0.4; CD5, p=0.4, CD3/CD5, p=0.2; CD8, p=0.8). Expression of L-selectin and ?2-integrins on granulocytes did not differ (CD11a, p=0.9; CD11b/c, p=0.7; CD62L, p=0.1). There were no differences in the percentage of apoptosis and necrosis (Annexin V, p=0.73; PI, p=0.21; Annexin V/PI, p=0.29). CONCLUSIONS: Data presented suggest that the down-regulation of the bone marrow triggered by BD is not related to changes in lymphocyte subsets, granulocyte adhesion molecules expression, or apoptosis and necrosis

Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-01-08T15:32:37Z No. of bitstreams: 2 Dissertação - Thamara Venâncio de Almeida - 2015.pdf: 1390852 bytes, checksum: 5f0b1c9700873dd65b4e3d743efec82f (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-01-11T06:39:56Z (GMT) No. of bitstreams: 2 Dissertação - Thamara Venâncio de Almeida - 2015.pdf: 1390852 bytes, checksum: 5f0b1c9700873dd65b4e3d743efec82f (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-01-11T06:39:56Z (GMT). No. of bitstreams: 2 Dissertação - Thamara Venâncio de Almeida - 2015.pdf: 1390852 bytes, checksum: 5f0b1c9700873dd65b4e3d743efec82f (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-08-21 / For assessing compliance with the quality requirements established by the Normative Instruction No. 62, a milk sample each property or community tank should be analyzed on a monthly basis by one of the laboratories accredited by the Brazilian Laboratories Network for Milk Quality Control . There are still doubts about what would be the maximum temperature and the time of milk sample storage without jeopardizing the results of the electronic analysis. The objective of this study was to evaluate the effects of temperature and storage time of cooled samples of raw milk on the results of milk quality electronic analysis. Refrigerated raw milk samples were collected from expansion tanks for individual use of farms located in the middle region of the state of Goiás, and stored at four different temperatures (3 °C, 11 °C, 17 °C and 25 °C) for 16 days. Total bacterial count (TBC), somatic cell count (SCC) and chemical composition were performed daily. The results were submitted to analysis of variance in split plot design in randomized blocks, and the means were compared by Duncan test. We concluded that, when azidiol is added as preservative to samples for TBC, they can be analyzed up to 16 days after collection when stored at temperatures of 3 °C and 11 °C, and up to 10 days when stored at 17 °C. Moreover, when bronopol is added as preservative, samples for SCC and chemical composition analysis can be stored for up to 16 days after collection when stored at temperatures of 3 °C and 11 °C, and for up to seven days when stored at 17 °C. / Para a avaliação do cumprimento dos requisitos de qualidade estabelecidos pela Instrução Normativa nº 62, uma amostra de leite de cada propriedade ou tanque comunitário deve ser analisada mensalmente por um dos laboratórios credenciados pela Rede Brasileira de Laboratórios de Controle da Qualidade do Leite. Ainda existem dúvidas sobre quais seriam a temperatura e o tempo máximos de armazenamento das amostras de leite sem que houvesse comprometimento dos resultados das análises eletrônicas. Objetivou-se com este trabalho avaliar os efeitos da temperatura e do tempo de armazenamento de amostras de leite cru refrigerado nos resultados das análises eletrônicas da qualidade do leite. Amostras de leite cru refrigerado foram coletadas de tanques de expansão de uso individual de fazendas localizadas na mesorregião Centro Goiano no Estado de Goiás e armazenadas em quatro temperaturas diferentes (3°C, 11°C, 17°C e 25°C) durante 16 dias. Foram realizadas diariamente análises de contagem bacteriana total (CBT), contagem de células somáticas (CCS) e composição química. Os resultados obtidos foram submetidos à análise de variância em esquema de parcelas subdivididas no tempo em delineamento de blocos ao acaso e as médias comparadas pelo teste de Duncan. Concluiu-se que as amostras destinadas à análise de CBT, adicionadas do conservante azidiol, podem ser analisadas por até 16 dias após a coleta quando armazenadas em temperaturas de 3°C e 11°C e por até 10 dias quando armazenadas a 17°C. E as amostras destinadas às análises de CCS e composição química, adicionadas do conservante bronopol, podem ser analisadas por até 16 dias após a coleta quando armazenadas em temperaturas de 3°C e 11°C e por até sete dias quando armazenadas a 17°C.

The concern about food safety, animal welfare, and environmental impacts caused by intensive production systems has led to an increasing interest in foods derived from a sustainable production system. In this context, organic milk production in Brazil has grown considerably in recent years, but little is known about the quality of the food produced in the country. In order to evaluate the quality of organic milk, the hygienic quality, chemical composition, and safety concerning pesticide residues was assessed. Five brands of certified organic pasteurized milk and five brands of conventional pasteurized milk commercialized in three Brazilian states were collected between May 2011 and March 2012, every two months, totaling 29 samples of organic milk (OM) and 27 conventional milk (CM). The collection of raw milk samples was made every two months, from July 2011 to May 2012 in 20 organic farms and 20 conventional farms in western Santa Catarina, totaling 120 OM and 115CM samples. In pasteurized milk, physicochemical analyzes were performed and the fatty acid profile was determined. In raw milk, chemical composition analyzes, somatic cell count (SCC), and total bacterial count (TBC) were carried out. Both types of milk were subjected to the determination of pesticides and veterinary drugs. When the results of raw milk production in the two systems were compared, no difference for the mean value of fat, protein and total solids, or SCC was found. Organic milk showed better quality in relation to microbial contamination and higher content of lactose. For pasteurized milk with the exception of non fat solids, all average values of physical and chemical parameters of milk met the limits of the Brazilian legislation. The fatty acid profile was affected by geographical origin, and to a lesser extent, the effect of season, probably due to differences on the feeding regime. The milk obtained in Rio Grande do Sul and São Paulo were characterized by having higher values of health-promoting fatty acids, including conjugated linoleic acid (CLA). However, the milk collected in the Federal District was characterized by higher concentrations of saturated fatty acids (SFA). The results for raw and pasteurized milk indicate that the organic management alone had no effect on milk quality regarding chemical composition, which are determined by several factors that are independent of the production system. No veterinary drugs was found in any of the samples analyzed. However, for contamination with pesticides no difference was observed when considering the number of contaminated samples in either production system. Results indicate the need to monitor milk for the presence of pesticides in order to improve food safety, especially for organic milk, which should be free of such contaminants. / A preocupação com segurança alimentar, bem-estar animal e impactos no meio ambiente causados pelos sistemas intensivos de produção, tem levado a um maior interesse por alimentos que sejam oriundos de um sistema de produção sustentável. Neste contexto, a produção de leite orgânico no Brasil cresceu consideravelmente nos últimos anos, porém pouco se sabe sobre a qualidade deste alimento produzido no país. Para avaliar a qualidade do leite orgânico foram realizadas análises que contemplaram aspectos da qualidade higiênica, composição química e a segurança quanto à presença de resíduos químicos. Cinco marcas de leite orgânico pasteurizado certificado e cinco de leite pasteurizado convencional comercializados em três Estados brasileiros foram coletadas entre maio de 2011 e março de 2012, com intervalo de dois meses, totalizando 29 amostras de leite orgânico (LO) e 27 de leite convencional (LC). As coletas de leite cru foram feitas de julho de 2011 a maio de 2012, também bimestrais, em 20 propriedades orgânicas e 20 propriedades convencionais na região Oeste de Santa Catarina, totalizando 120 amostras de LO e 115 de LC. No leite pasteurizado foram realizadas análises físico-químicas e o perfil de ácidos graxos. No leite cru foram realizadas as análises de composição química, contagem de células somáticas (CCS) e contagem bacteriana total (CBT). Ambos os tipos de leite foram submetidos à determinação de agrotóxicos e medicamentos veterinários. Comparando-se os resultados do leite cru, entre os dois sistemas de produção, não foi encontrado diferença para o valor médio de gordura, proteína e sólidos totais, assim como para CCS. O leite orgânico apresentou melhor qualidade em relação a contaminação microbiana e teor superior de lactose. Para o leite pasteurizado com exceção do extrato seco desengordurado (ESD), todos os valores médios dos parâmetros físicos e químicos atenderam aos limites da legislação brasileira. O perfil de ácidos graxos sofreu efeito da origem geográfica, e em menor escala o efeito da sazonalidade, provavelmente por diferenças no regime alimentar. Os leites obtidos no Rio Grande do Sul e em São Paulo se caracterizaram por possuir valores superiores de ácidos graxos promotores da saúde, entre eles o ácido linoléico conjugado (CLA), enquanto os leites coletados no Distrito Federal se caracterizaram por concentrações superiores de ácidos graxos saturados. Os resultados para o leite cru e pasteurizado indicam que o manejo orgânico por si só, não influenciou na qualidade do leite quanto aos aspectos de composição química, que são determinados por vários fatores que independem do sistema de produção. Não foram encontrados medicamentos veterinários em nenhuma das amostras analisadas. Quanto à contaminação por agrotóxicos não houve diferença quando considerado o número de amostras contaminadas em ambos os sistemas de produção. Os resultados indicam a necessidade de monitorar o leite quanto a presença de agrotóxicos a fim de melhorar a segurança alimentar, principalmente para leite orgânico, que deve estar livre desse tipo de contaminantes.