Com o objetivo de avaliar a função metabólica de vacas da raça Holandesa com diferentes graus de catarro genital, foram examinadas 230 vacas e foram colhidas 133 amostras de sangue de fêmeas no puerpério tardio, entre 15 e 42 dias. As amostras foram separadas em quatro grupos de animais, os que apresentaram catarro genital grau I, aqueles com catarro genital grau II, animais com catarro genital grau III e animais saudáveis que formaram o grupo controle, sendo dos grupos G1, G2, G3 e G4 respectivamente. Avaliou-se a função renal a partir da determinação dos teores séricos de uréia e creatinina, a função hepática através da determinação dos teores séricos de proteína total, de albumina, da atividade enzimática da aspartato aminotransferase (AST) e da gamaglutamiltransferase (GGT), o lipidograma foi avaliado por meio da determinação dos teores séricos de colesterol, de triglicérides e dos teores plasmáticos de ácidos graxos não esterificados (NEFA) e do betahidroxibutirato. Além disso foram avaliados os teores plasmáticos de glicose e séricos dos íons cálcio, fósforo e magnésio do soro. A determinação dos teores das variáveis estudadas foram quantificadas em analisador bioquímico automático da marca MAS®, modelo Liasys®. Os teores séricos de uréia foram estatisticamente diferentes entre os grupos 1 e 2. Os teores séricos de creatinina foram estatisticamente maiores nos animais doentes. Não houve diferença estatistica entre os teores de proteína sérica total nos diferentes grupos. Houve diferença estatistica entre os teores séricos de albumina entre os grupos 1 e 3. Houve diferença estatistica entre os teores séricos de AST entre os grupos 1 e 4. Não houve diferença estatistica entre os teores séricos de GGT. Houve diferença estatistica entre os teores séricos de colesterol entre os grupos. Não houve diferença estatistica entre os teores séricos de triglicérides entre os grupos. Não houve diferença estatistica entre os grupos em relação aos teores plasmáticos de NEFA, de β-Hidroxibutirato e de glicose. Houve diferença estatística entre os teores séricos de cálcio entre os grupos 1 e 4. Houve diferença estatística entre os teores séricos de fósforo entre os grupos 1 e 2 e entre os grupos 1 e 4. Houve diferença estatística entre os teores séricos de magnésio entre o grupo 1 e 4. / In order to evaluate the metabolic function of Holstein Friesian cows with different degrees of vaginal catarrh, 230 cows were examined and 133 blood samples were collected from late postpartum females between 15 and 42 days. The blood samples were separated into four experimental groups, animals with vaginal catarrh grade I, animals with vaginal catarrh grade II, animals with vaginal catarrh grade III and healthy animals comprising the control group, being G1, G2, G3 and G4, respectively. We evaluated renal function by determining the levels of serum urea and creatinine, liver function by determining the levels of serum total protein, albumin, the enzymatic activity of aspartate aminotransferase (AST) and gammaglutamyltransferase (GGT), the lipid profile was evaluated by determining the levels of serum cholesterol, triglyceride levels and plasma non-esterified fatty acids (NEFA) and β-hidroxibutyrate. In addition we evaluated the levels of glucose in plasma, calcium, magnesium and phosphorus in serum. The determination of the levels of variables were quantified with the use of automatic biochemical analyzer, AMS® brand, model Liasys®. The serum urea levels were statistically different between groups 1 and 2. The serum creatinine levels were statistically higher in diseased animals. There was no statistical difference between the levels of total serum protein in the different groups. There was statistical difference between serum albumin levels between groups 1 and 3. There was statistical difference between serum AST levels between groups 1 and 4. There was no statistical difference between the serum levels of GGT. There was statistical difference between serum cholesterol levels between the groups. There was no statistical difference between the serum triglyceride levels between the groups. There was no statistical difference between groups in relation to plasma levels of NEFA, β-hydroxybutyrate and glucose. There was statistical difference between the serum calcium levels between groups 1 and 4. There was statistical difference between serum phosphorus levels between groups 1 and 2 and between groups 1 and 4. There was statistical difference between serum magnesium levels between groups 1 and 4.

Do ponto de vista da produtividade, a fertilidade é um dos parâmetros de maior importância em um rebanho bovino comercial e esta é influenciada por vários fatores, entre eles estão as condições do trato reprodutivo das fêmeas e a qualidade do sêmen utilizado. O influxo de células inflamatórias no local da deposição do sêmen logo após a inseminação artificial (IA) pode ser intensificada na presença de maior número de espermatozoides lesados durante a IA, caracterizando uma endometrite. Este estudo foi conduzido em três experimentos. Com o objetivo de comparar os métodos de colheita de material endometrial por escova ginecológica (EU) e lavado uterino (LU), bem como a interferência destes procedimentos na hemodinâmica uterina, foi proposto o Experimento 01, onde pode-se constatar que ambas as técnicas permitem o recolhimento de amostras em quantidade e qualidade suficiente para contagem, e que a porcentagem de células polimorfonucleares obtidas pela técnica LU foi superior a EU. Maior fluxo sanguíneo das artérias uterinas foi encontrado no momento de 4 horas após a realização de LU, sugerindo que este influencia na resposta vascular inflamatória. Para avaliar o efeito da LU após a IA em Tempo Fixo (IATF) na fertilidade dos animais, executou-se o Experimento 02 e constatou-se que não há diferença no índice de prenhez entre os animais submetidos ou não à LU, demonstrando que a técnica não interfere na taxa de fertilidade. Com o intuito de investigar a interferência da qualidade do sêmen na fertilidade, resposta inflamatória e hemodinâmica uterina, foi proposto o Experimento 03, onde foi possível observar influência da qualidade do sêmen sobre a taxa de prenhez, verificou-se maior porcentagem de vacas prenhes quando inseminadas com sêmen com maiores percentuais de espermatozoides apresentando integridade das membranas plasmática e acrossomal e função mitocondrial (PIAIC). Notou-se ainda a ocorrência de endometrite em 65,3 % dos animais, os quais apresentaram taxa de prenhez inferior aos que não apresentaram inflamação. Pode-se concluir que a qualidade do sêmen e a endometrite interferem na taxa de fertilidade bovina. / When productivity is taking into account, fertility is one of the most important parameters in a commercial herd. It is influenced by several factors, especially by the conditions of the female reproductive tract and the quality of the semen used. The influx of inflammatory cells at the site of semen deposition after artificial insemination (AI) can be intensified by the deposition of a greater number of dead spermatozoa during AI, which characterized endometritis. This study was conducted in three different experiments. In order to compare the methods of collection of endometrial sampling by swab using a gynecological brush (GB) or uterine flushing (UF), as well as the interference of these procedures in uterine hemodynamics, we designed experiment 01. Our results reveal that both techniques allow collecting samples with good quality and sufficient quantity to be counted; moreover, the average percentage of polymorphonuclear cells obtained by UF was greater compared to those obtained by GB. It may be noted that the increased blood flow was observed in samples collected four hours after the UF procedure, suggesting that it may have an influence on the vascular inflammatory response. To evaluate the effect of uterine flushing after AIFT on animal fertility we designed the experiment 02. Our results revealed that there is no statistical difference in pregnancy rates between flushed and non flushed animals, showing that the UF does not interfere with fertility rate. Experiment 03 was designed in order to assess the inflammatory response induced by different qualities of semen and their interference on uterine hemodynamic and fertility. There was an influence of semen quality on pregnancy rates: higher percentage of pregnancy was found in the group of cows inseminated with semen with plasma and acrossome membrane integrity and mitochondrial function (PIAIC). Endometritis was noticed in 65.3% of the cows and these animals presented lower pregnancy rate compared to those that did not show an inflammatory response. We concluded that semen quality and endometritis interferes with fertility rate in bovine species.

L’endométrite est une maladie affectant les performances en reproduction chez la vache laitière. Elle est définie par la présence d’inflammation excessive au niveau de l’utérus durant la période post-partum. Le traitement antibiotique sous forme d’infusion intra-utérine est l’une des approches curatives les plus utilisées et améliore les performances de reproduction subséquentes. Considérant les enjeux modernes en lien avec l’antibiorésistance et la réduction de l’utilisation d’antibiotiques en productions animales, il est justifiable de remettre en question l’utilisation d’une méthode diagnostique de l’endométrite basée uniquement sur la présence d’inflammation en vue de déterminer si un traitement antibiotique doit être utilisé. En ce sens, l’objectif principal de ce projet de recherche était de valider l’exactitude des résultats de milieux de culture bactériologique utilisés à la ferme (Tri-plate et Petrifilm) en se basant sur ceux obtenus en laboratoire diagnostique. Un objectif secondaire était de vérifier la concordance entre les résultats bactériens de laboratoire et ceux cytologiques. Pour ce faire, une étude observationnelle transversale a été réalisée au sein de deux troupeaux laitiers commerciaux faisant partie de la clientèle de la Clinique ambulatoire bovine de la Faculté de médecine vétérinaire de l’Université de Montréal. Un total de 189 vaches en période post-partum (30 à 43 jours de lactation) ont été enrôlées de façon systématique afin de recueillir 2 échantillons utérins à partir de cytobrosses lors d’un même examen. La première cytobrosse était utilisée pour inoculer directement le milieu de culture Tri-plate et ensuite être envoyée au laboratoire du Service diagnostic de la FMV de l’Université de Montréal pour culture bactérienne aérobique. La seconde cytobrosse était utilisée pour l’analyse cytologique (proportion de cellules polymorphonucléaires; PMNL), puis était diluée dans 1 ml de solution saline pour inoculer le milieu Petrifilm. Après une période d’incubation de 24h, le décompte de colonies était fait pour chaque milieu de culture. À partir de ces données, les analyses statistiques ont été complétées dans le but d’optimiser la sommation de sensibilité et spécificité (Se+Sp) des deux milieux de culture en fonction des résultats du laboratoire de référence. Pour le milieu Tri-plate, le seuil de ˃ 90 colonies a permis d’obtenir la Se+Sp maximale, soit 167,7. Pour le milieu Petrifilm, le seuil de ˃100 colonies a donné la Se+Sp maximale, soit 129. En ce qui concerne la concordance entre le diagnostic cytologique et bactériologique, les résultats de notre projet montrent que 64,3% des vaches positives au niveau cytologique (˃6% PMNL) se sont avérées négatives à la culture bactérienne, démontrant un manque de concordance. En conclusion, les résultats de notre étude supportent la faisabilité et le potentiel de la culture bactérienne d’échantillons de l’endomètre à la ferme. Notre projet met également en évidence que l’utilisation d’un dépistage cytologique de l’endométrite afin d’utiliser un traitement antibiotique puisse causer un manque d’optimisation dans l’efficacité des traitements. Il semble donc pertinent que d’autres projets soient complétés afin d’approfondir le dépistage bactériologique de l’endométrite et d’analyser le potentiel diagnostique de celui-ci. / Endometritis is a disease affecting reproductive performance in dairy cows. It is defined by the presence of excessive inflammation in the uterine body during the postpartum period. From a curative point of view, the use of antibiotic treatment in the form of intrauterine infusion is one of the most widely used methods which has a beneficial effect on subsequent reproductive performance in positive cases of endometritis. Considering the modern issues concerning the use of antibiotics in animal production, the use of a diagnostic method based on the presence of inflammation in order to administer antibiotic treatment is questionable. In that sense, the main objective of this research project was to validate the accuracy of the results of bacteriological culture medium used on the farm (Tri-plate and Petrifilm) based on the ones from diagnosis laboratory. A secondary objective was to verify the concordance between the bacterial and cytological results. To do this, a cross-sectional observational study was set up within two commercial dairy herds followed by the bovine outpatient clinic of the Faculty of Veterinary Medicine of the Université de Montréal. A total of 189 cows in the postpartum period (30 to 43 days in milk) were systematically enrolled in order to collect 2 uterine samples from cytobrush during the same examination. The first cytobrush was used to directly inoculate the Tri-plate medium and then be sent to the reference laboratory (Diagnostic service of the FVM of the Université de Montréal) for aerobic bacterial analysis. The second cytobrush was initially used to make a microscopic smear for cytological analysis (polymorphonuclear cell proportion (PMNL)) and subsequently diluted in 1 ml of saline to inoculate the Petrifilm medium. After an incubation period of 24 hours, the colony count was made for each culture medium. From these data, statistical analyses were completed in order to optimize the summation of sensitivity and specificity (Se + Sp) of the two culture medium according to the results of the reference laboratory. For the Tri-plate medium, the cutoff of ˃ 90 colonies resulted in the maximum Se + Sp, which was 167.7. For the Petrifilm medium, the threshold of ˃100 colonies gave the maximum Se + Sp, which was 129. Regarding the concordance between the cytological and bacterial diagnosis, the results of our project show that 64.3% of cows positive for cytological criteria (˃6% PMNL) were found to be negative from a bacterial point of view. In conclusion, this study shows that Tri-plate and Petrifilm medium used on-farms were best to reproduce the results obtained by laboratory analysis using 90 and 100 colony threshold respectively. Our results also support a lack of agreement between cytological and bacterial diagnosis. It therefore seems relevant that other projects are completed in order to deepen the bacterial screening for endometritis and to analyze its diagnostic potential.

L’objectif de la présente étude était d’évaluer un test d’estérase leucocytaire (LE) pour le diagnostic de l’endométrite subclinique chez les vaches Holstein en période postpartum. Les tests effectués à partir d’échantillons provenant soit de l’endomètre (UtLE) ou du col utérin (CxLE) ont été comparés à la cytologie endométriale (CE). Par ailleurs, deux méthodes d’évaluation des lames ont été comparées. Deux cent quatre vingt-cinq vaches Holstein de 5 troupeaux laitiers commerciaux ont été évaluées entre 21 et 47 jours en lait (JEL). Soixante sept vaches ont été diagnostiquées avec une endométrite clinique suite à un examen transrectal et vaginoscopique et ont été exclues de l’étude. Deux cent dix-huit vaches ont eu des prélèvements pour la CE et le test LE. La fonction ovarienne a été déterminée à la palpation transrectale. La banque de données utilisée pour chacune des vaches a été effectuée à partir du logiciel DSA (Dossier de Santé Animale) laitier. Le pourcentage de neutrophiles était significativement corrélé avec les scores de LE utérin et cervical. L’activité de CxLE et UtLE diminuait significativement avec les JEL, mais n’était pas associée au risque de gestation à 90 JEL (n= 186). Le pourcentage de neutrophiles mesuré à la CE entre 32 et 47 JEL était associé significativement au risque de gestation à 90 JEL (n=94, P=0.04). Pour la même période, selon une analyse de survie, les vaches avec >2,6% de neutrophiles à la CE étaient définies comme étant atteintes d’une endométrite subclinique avec une prévalence de 56%. Les résultats indiquent que le test d’estérase utérin ou cervical a une bonne concordance avec le pourcentage de neutrophiles à la CE. Une endométrite subclinique diagnostiquée par cytologie endometriale entre 32 et 47 JEL est associée à une réduction du risque de gestation au premier service. / The point toward this study was to determine the diagnostic test characteristics of the leukocyte esterase activity test for subclinical endometritis in postpartum Holstein dairy cows. The objectives were 1) to compare uterine leukocyte esterase activity and the endometrial cytology (EC), 2) to compare leukocyte esterase activity of the cervix (CxLE) and the uterus (UtLE), 3) Compare two methods of assessing the slides (i.e. an exhaustive method and a rapid method). Two hundred eighty five post partum Holstein cows from 5 commercial dairy herds had a post partum evaluation between 21 and 47 days in milk (DIM). Sixty seven cows where diagnosed with clinical endometritis by transrectal and vaginoscopy examinations and were excluded from the study. Two hundred eighteen cows were enrolled for endometrial cytology and esterase activity test. The ovarian status was determined by transrectal examination. Computerized databank, dairy DSA (Dossier de Santé Animale) indexing all the cows was used to retrieve individual information for analysis. The percentage of neutrophils was significantly correlated with the LE from the uterus and cervix. The LE from cervix and uterus decreased significantly with DIM, however, they were not statistically associated with pregnancy risk at 90 DIM (n=186). Between 32-47 DIM, the percentage of neutrophils and risk of pregnancy at 90 DIM were associated (n=94, P=0.04). For the same period, survival analysis identified cows with > 2.6 % neutrophils on EC as subclinical endometritis cows with a prevalence of 56%. The two methods for assessing the slides were correlated by 81%. Subclinical endometritis diagnosed by endometrial cytology between 32 and 47 DIM was associated with reduced risk of pregnancy at first service.

Após a inseminação artificial, as fêmeas apresentam reação inflamatória transitória e fisiológica, que pode resultar em alterações na vascularização uterina. A ultrassonografia Doppler colorido é uma técnica não invasiva, que vem sendo utilizada para estudar a hemodinâmica do trato reprodutivo de bovinos. Os objetivos deste trabalho foram verificar a resposta inflamatória uterina por ultrassonografia Doppler colorido e a taxa de fertilidade em bovinos após inseminação artificial (IA) com sêmen avaliado por sondas fluorescentes. Para isso foram delineados dois experimentos. O experimento 1 foi realizado para estudar a hemodinâmica uterina por ultrassonografia Doppler colorido em bovinos após a IA. Foram utilizadas 18 vacas da raça Nelore, submetidas a um protocolo para a inseminação artificial em tempo fixo (IATF) e divididas em dois grupos: Grupo não inseminado (controle, GC, n=9) e grupo submetido à inseminação artificial (GIA, n=9). Avaliações por ultrassonografia foram realizadas em cinco momentos em modo espectral (30 horas antes da IA, 4, 24, 48 e 168 horas após a IA) para avaliar o índice de resistência (RI) da artéria uterina e em quatro momentos em modo color Doppler (4, 24, 48 e 168 horas após a IA) para avaliar a vascularização dos cornos uterinos. O diagnóstico de gestação foi feito após 30 dias da IA. No experimento 2 objetivou-se verificar se o percentual de espermatozóides com integridade das membranas plasmática e acrossomal e função mitocondrial (PIAIC) avaliadas por sondas fluorescentes influencia a fertilidade e estudar a hemodinâmica uterina após a deposição do sêmen com diferentes percentuais de espermatozóides PIAIC. Foram utilizadas 182 vacas paridas da raça Nelore, submetidas a um protocolo para IATF e divididas em três grupos de acordo com a qualidade do sêmen: Bom (B) 44,5%, Médio (M) 23% e Regular (R) 8,5% de espermatozóides PIAIC/palheta. Uma amostra de 30% das fêmeas (54, sendo 18 de cada grupo) foram submetidas a avaliações com ultrassonografia Doppler colorido 30 horas antes da IA, 4 e 24 horas após a IA. Foram considerados os RI das artérias uterinas direita (AD) e esquerda (AE) e vascularização subjetiva dos cornos uterinos direito (CD) e esquerdo (CE). Posteriormente, foram separados levando em consideração o lado do ovário contendo o folículo dominante para comparar RI das artérias ipsi (AI) e contralateral (AC) e vascularização subjetiva dos cornos uterinos ipsi (CI) e contralateral (CC). O diagnóstico de gestação foi realizado 30 dias após a IA. Para a análise estatística foi utilizado o programa SAS versão 9.2 (SAS 2010). As variáveis foram analisadas pelo PROC MIXED, utilizando modelo linear misto para medidas repetidas no tempo. Foram considerados os efeitos fixos de grupo, tempo e interação grupo x tempo. Os dados obtidos foram submetidos à análise de variância e ao teste de Duncan, pelo PROC GLM. A análise de Qui-Quadrado foi utilizada para determinar a taxa de prenhez, de acordo com a qualidade do sêmen pelo PROC FREQ. O nível de significância considerado foi de 5%. No experimento 1 não foram observadas diferenças dos parâmetros avaliados por ultrassom Doppler entre GIA e GC, mas foram notados efeitos do tempo para RI, sendo mais baixos 4 horas após a IA. Quando os dados foram avaliados de acordo com o lado do folículo dominante foi observado que 4 e 24 horas após a IA, RI da AC do GIA foi mais baixo do que o GC. No experimento 2 não houve diferença entre os animais inseminados com sêmen B, M ou R quanto a hemodinâmica uterina, mas houve efeito do tempo, sendo que 4 horas após a IA verificou-se menor valor de RI e maior fluxo sanguíneo. A taxa de prenhez dos animais inseminados com o sêmen B (64,7%) foi maior quando comparada ao sêmen R (36,2%), sendo que ambos não diferiram do sêmen M (50,0%). Pode-se concluir que não é possível detectar por ultrassonografia Doppler alterações na hemodinâmica uterina devido à IA em fêmeas bovinas. A deposição de sêmen com diferentes qualidades não altera a hemodinâmica uterina de forma detectável por ultrassonografia Doppler em bovinos. Por outro lado, é possível detectar por ultrassonografia Doppler mudanças na perfusão sanguínea do útero em relação ao tempo da IA, o que pode ser devido a alterações hormonais que ocorrem durante o ciclo estral. Por fim, a IA com sêmen contendo menor percentual de espermatozoides PIAIC causa redução na taxa de fertilidade em bovinos. / After insemination, females have physiological and transient inflammatory reaction, which can result in changes in the uterine vasculature. The color Doppler ultrasound is a noninvasive technique that has been used to study the hemodynamics of cattle reproductive tract.The purposes of this work were to verify uterine inflammatory response by color Doppler ultrasound and fertility rate in cattle after artificial insemination (IA) with semen evaluated by fluorescent probes. For this, two experiments were delineated. The first experiment was conducted to study the uterine hemodynamic by color Doppler ultrasound in cattle after AI. Were used 18 Nellore cows that were subjected to a protocol for fixed timed artificial insemination (IATF). The animals were divided in two groups: Not inseminated group (control, GC, n=9) and inseminated group (GIA, n=9). Ultrasonografic evaluation were done in five moments on spectral mode (30 hours before AI, 4, 24, 48 e 168 hours after IA) to evaluate the resistence index (RI) of uterine artery and in four moments in color Doppler mode to evaluate the uterine horns vascularization. The pregnancy diagnostic was done 30 days after IA. In the second experiment the objective was to evaluate whether the percentage of sperm with plasma membrane integrity and acrossomal and mitochondrial function (PIAIC) assessed by fluorescent probes influences fertility and study the uterine hemodynamic after the deposition of semen with different percentages of PIAIC sperm. Were used 182 caved Nellore cows submitted to a protocol for IATF and divided into three groups according to semen quality: Good (B) 44.5%, Medium (M) 23% and Regular (R) 8,5% of PIAIC sperm/blade. A sample of 30% of female (54, 18 in each group) were submitted to evaluation by color Doppler ultrasound 30 hours before IA, 4 and 24 hours after IA. Were considered RI of right (AD) and left (AE) uterine arteries and subjective vascularization of right (CD) and left (CE) uterine horns. Later, they were separated taking into account the side of the ovary containing the dominant follicle to compare RI of ipsi (AI) and contralateral (AC) arteries and subjective vascularization of ipsi (CI) and contralateral (CC) uterine horns. The pregnancy diagnostic was done 30 days after IA. Statistical analysis was performed using the program SAS version 9.2 (SAS 2010). The variables were analyzed by PROC MIXED, using mixed linear model for repeated measures in time. We considered effects of group, time and group x time interaction. The data obtained were submitted to ANOVA and Duncan test, using PROC GLM. The chi-square analysis was used to determine pregnancy rate, according to the quality of semen by PROC FREQ. the significance level was 5%. In experiment 1 there were no differences of the parameters evaluated by Doppler ultrasound between GIA and GC, but time effects were noted for RI, it was lower four hours after IA. When the data were evaluated according to the side of the dominant follicle was observed that 4 and 24 hours after IA, RI of AC of GIA was lower when it was compared with GC. In experiment 2 there was no difference between animals inseminated with semen B, M or R about uterine hemodynamics, but there were effects of time, so that 4 hours after IA showed lower value of RI and increased blood flow. The pregnancy rate of animals inseminated with semen B (64.7%) was higher when compared with semen R (36.2%), both of them did not differ of semen M (50.0%). It can be concluded that is not possible to detect uterine hemodynamic changes by Doppler ultrasound due to IA in cattle. The deposition of semen with different qualities does not change the uterine hemodynamics enough to be detectable by Doppler ultrasonography in cattle. On the other hand, is possible to detect changes in blood perfusion of the uterus in relation to the time of IA by Doppler ultrasound, which may be due to hormonal changes that occur during the estrous cycle. Finally, IA with semen containing a lower percentage of PIAIC sperm causes a reduction in fertility rate in cattle.

Após a inseminação artificial, as fêmeas apresentam reação inflamatória transitória e fisiológica, que pode resultar em alterações na vascularização uterina. A ultrassonografia Doppler colorido é uma técnica não invasiva, que vem sendo utilizada para estudar a hemodinâmica do trato reprodutivo de bovinos. Os objetivos deste trabalho foram verificar a resposta inflamatória uterina por ultrassonografia Doppler colorido e a taxa de fertilidade em bovinos após inseminação artificial (IA) com sêmen avaliado por sondas fluorescentes. Para isso foram delineados dois experimentos. O experimento 1 foi realizado para estudar a hemodinâmica uterina por ultrassonografia Doppler colorido em bovinos após a IA. Foram utilizadas 18 vacas da raça Nelore, submetidas a um protocolo para a inseminação artificial em tempo fixo (IATF) e divididas em dois grupos: Grupo não inseminado (controle, GC, n=9) e grupo submetido à inseminação artificial (GIA, n=9). Avaliações por ultrassonografia foram realizadas em cinco momentos em modo espectral (30 horas antes da IA, 4, 24, 48 e 168 horas após a IA) para avaliar o índice de resistência (RI) da artéria uterina e em quatro momentos em modo color Doppler (4, 24, 48 e 168 horas após a IA) para avaliar a vascularização dos cornos uterinos. O diagnóstico de gestação foi feito após 30 dias da IA. No experimento 2 objetivou-se verificar se o percentual de espermatozóides com integridade das membranas plasmática e acrossomal e função mitocondrial (PIAIC) avaliadas por sondas fluorescentes influencia a fertilidade e estudar a hemodinâmica uterina após a deposição do sêmen com diferentes percentuais de espermatozóides PIAIC. Foram utilizadas 182 vacas paridas da raça Nelore, submetidas a um protocolo para IATF e divididas em três grupos de acordo com a qualidade do sêmen: Bom (B) 44,5%, Médio (M) 23% e Regular (R) 8,5% de espermatozóides PIAIC/palheta. Uma amostra de 30% das fêmeas (54, sendo 18 de cada grupo) foram submetidas a avaliações com ultrassonografia Doppler colorido 30 horas antes da IA, 4 e 24 horas após a IA. Foram considerados os RI das artérias uterinas direita (AD) e esquerda (AE) e vascularização subjetiva dos cornos uterinos direito (CD) e esquerdo (CE). Posteriormente, foram separados levando em consideração o lado do ovário contendo o folículo dominante para comparar RI das artérias ipsi (AI) e contralateral (AC) e vascularização subjetiva dos cornos uterinos ipsi (CI) e contralateral (CC). O diagnóstico de gestação foi realizado 30 dias após a IA. Para a análise estatística foi utilizado o programa SAS versão 9.2 (SAS 2010). As variáveis foram analisadas pelo PROC MIXED, utilizando modelo linear misto para medidas repetidas no tempo. Foram considerados os efeitos fixos de grupo, tempo e interação grupo x tempo. Os dados obtidos foram submetidos à análise de variância e ao teste de Duncan, pelo PROC GLM. A análise de Qui-Quadrado foi utilizada para determinar a taxa de prenhez, de acordo com a qualidade do sêmen pelo PROC FREQ. O nível de significância considerado foi de 5%. No experimento 1 não foram observadas diferenças dos parâmetros avaliados por ultrassom Doppler entre GIA e GC, mas foram notados efeitos do tempo para RI, sendo mais baixos 4 horas após a IA. Quando os dados foram avaliados de acordo com o lado do folículo dominante foi observado que 4 e 24 horas após a IA, RI da AC do GIA foi mais baixo do que o GC. No experimento 2 não houve diferença entre os animais inseminados com sêmen B, M ou R quanto a hemodinâmica uterina, mas houve efeito do tempo, sendo que 4 horas após a IA verificou-se menor valor de RI e maior fluxo sanguíneo. A taxa de prenhez dos animais inseminados com o sêmen B (64,7%) foi maior quando comparada ao sêmen R (36,2%), sendo que ambos não diferiram do sêmen M (50,0%). Pode-se concluir que não é possível detectar por ultrassonografia Doppler alterações na hemodinâmica uterina devido à IA em fêmeas bovinas. A deposição de sêmen com diferentes qualidades não altera a hemodinâmica uterina de forma detectável por ultrassonografia Doppler em bovinos. Por outro lado, é possível detectar por ultrassonografia Doppler mudanças na perfusão sanguínea do útero em relação ao tempo da IA, o que pode ser devido a alterações hormonais que ocorrem durante o ciclo estral. Por fim, a IA com sêmen contendo menor percentual de espermatozoides PIAIC causa redução na taxa de fertilidade em bovinos. / After insemination, females have physiological and transient inflammatory reaction, which can result in changes in the uterine vasculature. The color Doppler ultrasound is a noninvasive technique that has been used to study the hemodynamics of cattle reproductive tract.The purposes of this work were to verify uterine inflammatory response by color Doppler ultrasound and fertility rate in cattle after artificial insemination (IA) with semen evaluated by fluorescent probes. For this, two experiments were delineated. The first experiment was conducted to study the uterine hemodynamic by color Doppler ultrasound in cattle after AI. Were used 18 Nellore cows that were subjected to a protocol for fixed timed artificial insemination (IATF). The animals were divided in two groups: Not inseminated group (control, GC, n=9) and inseminated group (GIA, n=9). Ultrasonografic evaluation were done in five moments on spectral mode (30 hours before AI, 4, 24, 48 e 168 hours after IA) to evaluate the resistence index (RI) of uterine artery and in four moments in color Doppler mode to evaluate the uterine horns vascularization. The pregnancy diagnostic was done 30 days after IA. In the second experiment the objective was to evaluate whether the percentage of sperm with plasma membrane integrity and acrossomal and mitochondrial function (PIAIC) assessed by fluorescent probes influences fertility and study the uterine hemodynamic after the deposition of semen with different percentages of PIAIC sperm. Were used 182 caved Nellore cows submitted to a protocol for IATF and divided into three groups according to semen quality: Good (B) 44.5%, Medium (M) 23% and Regular (R) 8,5% of PIAIC sperm/blade. A sample of 30% of female (54, 18 in each group) were submitted to evaluation by color Doppler ultrasound 30 hours before IA, 4 and 24 hours after IA. Were considered RI of right (AD) and left (AE) uterine arteries and subjective vascularization of right (CD) and left (CE) uterine horns. Later, they were separated taking into account the side of the ovary containing the dominant follicle to compare RI of ipsi (AI) and contralateral (AC) arteries and subjective vascularization of ipsi (CI) and contralateral (CC) uterine horns. The pregnancy diagnostic was done 30 days after IA. Statistical analysis was performed using the program SAS version 9.2 (SAS 2010). The variables were analyzed by PROC MIXED, using mixed linear model for repeated measures in time. We considered effects of group, time and group x time interaction. The data obtained were submitted to ANOVA and Duncan test, using PROC GLM. The chi-square analysis was used to determine pregnancy rate, according to the quality of semen by PROC FREQ. the significance level was 5%. In experiment 1 there were no differences of the parameters evaluated by Doppler ultrasound between GIA and GC, but time effects were noted for RI, it was lower four hours after IA. When the data were evaluated according to the side of the dominant follicle was observed that 4 and 24 hours after IA, RI of AC of GIA was lower when it was compared with GC. In experiment 2 there was no difference between animals inseminated with semen B, M or R about uterine hemodynamics, but there were effects of time, so that 4 hours after IA showed lower value of RI and increased blood flow. The pregnancy rate of animals inseminated with semen B (64.7%) was higher when compared with semen R (36.2%), both of them did not differ of semen M (50.0%). It can be concluded that is not possible to detect uterine hemodynamic changes by Doppler ultrasound due to IA in cattle. The deposition of semen with different qualities does not change the uterine hemodynamics enough to be detectable by Doppler ultrasonography in cattle. On the other hand, is possible to detect changes in blood perfusion of the uterus in relation to the time of IA by Doppler ultrasound, which may be due to hormonal changes that occur during the estrous cycle. Finally, IA with semen containing a lower percentage of PIAIC sperm causes a reduction in fertility rate in cattle.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-08-28T19:02:05Z No. of bitstreams: 1 2017 - Marcus Andr? Ferreira S?.pdf: 1043888 bytes, checksum: 7a3d94bc8171597abb3f0dde51169347 (MD5) / Made available in DSpace on 2018-08-28T19:02:05Z (GMT). No. of bitstreams: 1 2017 - Marcus Andr? Ferreira S?.pdf: 1043888 bytes, checksum: 7a3d94bc8171597abb3f0dde51169347 (MD5) Previous issue date: 2017-08-23 / Objectives of this study were to characterize the endometrial vascular perfusion of mares with bacterial endometritis using color Doppler ultrasonography and submitted to phytotherapeutic treatment. 19 crossbred and Mangalarga Marchador mares without endometritis were used. ImageJ 1.46r? was applied to evaluate the images. Additionally, subjective vascular uterine perfusion was estimated analyzing the percentage of color Doppler signals present in the mesometrial, myometrium and endometrium, in longitudinal section of the uterine body and transverse section of the uterine horns. Step 1. The mares (n=20) were submitted to intrauterine inoculation with Escherichia coli cepa from the equine uterus. The color Doppler ultrasound sessions were performed in M0 (moment immediately prior to intrauterine inoculation) and M1 (24 hours after intrauterine inoculation). Step 2. The animals were divided into two groups: control group (n=10) and treated group (n=10) using phytotherapic solution Fitoclean? (Organnact Animal Health, Curitiba, Paran?, Brazil). Both groups, color Doppler ultrasonography was performed at time T1 (immediately before starting treatment), T2 (24h after treatment) and T3 (48h after treatment). Step 3. Mares in which pathogens were identified in the samples collected during stage 2 were submitted to antibiotic therapy by intrauterine infusion using Gentamicin. Seven days after the antibiotic therapy, a new color Doppler ultrasound examination was performed. For statistical analysis, the Tukey test, t Student and Anova test was applied to compare the means obtained in the different periods and Chi Square were apllied to evaluation of phytoterapic effect. Step 1. It was found that the mean values of vascularization were no moment (P<0.05). Bacterial growth was observed in all samples collected. Step 2. The mean value of vascularization at time T1 in both groups was significantly higher (P<0.05) than the obtained at moments 2 and 3. At the time T1, we observed a significant increase in portion 1, while T2 showed a significant decreasing in this portion. Step 3. According to the results obtained in the culture and antibiogram examinations performed during the step 2, 13 mares (65%, 13/20) were submitted to antibiotic therapy through intrauterine infusion with Gentamicin. After that treatment, the vascularization observed decreased related to M1. With these results, we can conclude that in these conditions is possible the use of color Doppler ultrasonography as a method for bacterial endometritis; It was not possible to correlate results obtained by color Doppler ultrasonography with the traditional findings for the diagnosis of endometritis. / Os objetivos do presente estudo foram caracterizar atrav?s da ultrassonografia color Doppler a perfus?o vascular endometrial de ?guas com endometrite bacteriana e submetidas a tratamento fitoter?pico. Para tanto, foram utilizadas 20 ?guas mesti?a e Mangalarga Marchador livres de endometrite. O programa ImageJ 1.46r? foi utilizado para avalia??o das imagens. Adicionalmente, a perfus?o vascular subjetiva do ?tero foi estimada levando-se em considera??o o percentual de sinais Doppler coloridos presentes no mesom?trio, miom?trio e endom?trio, em corte longitudinal do corpo uterino e transversal dos cornos uterinos. Etapa 1. Todas as ?guas (n=20) foram submetidas a inocula??o intrauterina com cepa de Escherichia coli proveniente de ?tero equino. As sess?es de ultrassonografia color Doppler foram realizadas em M0 (momento imediatamente anterior ? inocula??o intrauterina) e M1 (24 horas ap?s a inocula??o intrauterina). Etapa 2. Os animais foram divididos aleatoriamente em dois grupos: grupo experimental 1 (n=10) e grupo experimental 2 (n=10) com solu??o fitoter?pica Fitoclean? (Organnact Sa?de Animal, Curitiba, Paran?, Brasil). Em ambos os grupos, os exames de cultura uterina, antibiograma, citologia endometrial e ultrassonografia modo B e color Doppler foram realizados nos momentos T1 (imediatamente antes de iniciar o tratamento), T2 (24h ap?s o tratamento) e T3 (48h ap?s o tratamento). Etapa 3. As ?guas em que foram identificados agentes patog?nicos nas amostras coletadas durante a etapa 2, foram submetidas ? antibioticoterapia por infus?o intrauterina utilizando Gentamicina. Sete dias ap?s a realiza??o da antibioticoterapia, foi realizado novos exame de ultrassonografia color Doppler. Para an?lise estat?stica, foi aplicado o teste de Tukey, teste t de Student e Anova para compara??o das m?dias obtidas nos diferentes per?odos e Qui Quadrado para avalia??o do efeito do fitoter?pico. Etapa 1. Verificou-se que os valores m?dios de vasculariza??o no momento M1 foram significativamente superiores aos obtidos em M0 para as tr?s partes do ?tero (P<0,05). Houve crescimento bacteriano em todas as amostras coletadas. Etapa 2. O valor m?dio de vasculariza??o no momento T1 em ambos os grupos foi significativamente superior (P<0,05) aos obtidos nos momentos 2 e 3. No momento T1, observou-se aumento significativo no segmento 1, enquanto que em T2 observou-se uma redu??o significativa neste segmento. Etapa 3. De acordo com os resultados obtidos nos exames de cultura e antibiograma realizados ap?s o t?rmino da etapa 2, 13 ?guas (65%, 13/20) foram submetidas a antibioticoterapia atrav?s de infus?o intrauterina com Gentamicina. Ap?s este tratamento, a vasculariza??o observada apresentou grande redu??o em compara??o ao momento M1. Diante destes resultados, concluiu-se que nas condi??es deste estudo foi poss?vel utilizar a ultrassonografia modo Color Doppler como m?todo diagn?stico para a endometrite bacteriana; n?o foi poss?vel correlacionar resultados obtidos atrav?s da ultrassonografia color Doppler com os achado nos exames tradicionais para diagn?stico da endometrite.

L’objectif de la présente étude était d’évaluer un test d’estérase leucocytaire (LE) pour le diagnostic de l’endométrite subclinique chez les vaches Holstein en période postpartum. Les tests effectués à partir d’échantillons provenant soit de l’endomètre (UtLE) ou du col utérin (CxLE) ont été comparés à la cytologie endométriale (CE). Par ailleurs, deux méthodes d’évaluation des lames ont été comparées. Deux cent quatre vingt-cinq vaches Holstein de 5 troupeaux laitiers commerciaux ont été évaluées entre 21 et 47 jours en lait (JEL). Soixante sept vaches ont été diagnostiquées avec une endométrite clinique suite à un examen transrectal et vaginoscopique et ont été exclues de l’étude. Deux cent dix-huit vaches ont eu des prélèvements pour la CE et le test LE. La fonction ovarienne a été déterminée à la palpation transrectale. La banque de données utilisée pour chacune des vaches a été effectuée à partir du logiciel DSA (Dossier de Santé Animale) laitier. Le pourcentage de neutrophiles était significativement corrélé avec les scores de LE utérin et cervical. L’activité de CxLE et UtLE diminuait significativement avec les JEL, mais n’était pas associée au risque de gestation à 90 JEL (n= 186). Le pourcentage de neutrophiles mesuré à la CE entre 32 et 47 JEL était associé significativement au risque de gestation à 90 JEL (n=94, P=0.04). Pour la même période, selon une analyse de survie, les vaches avec >2,6% de neutrophiles à la CE étaient définies comme étant atteintes d’une endométrite subclinique avec une prévalence de 56%. Les résultats indiquent que le test d’estérase utérin ou cervical a une bonne concordance avec le pourcentage de neutrophiles à la CE. Une endométrite subclinique diagnostiquée par cytologie endometriale entre 32 et 47 JEL est associée à une réduction du risque de gestation au premier service. / The point toward this study was to determine the diagnostic test characteristics of the leukocyte esterase activity test for subclinical endometritis in postpartum Holstein dairy cows. The objectives were 1) to compare uterine leukocyte esterase activity and the endometrial cytology (EC), 2) to compare leukocyte esterase activity of the cervix (CxLE) and the uterus (UtLE), 3) Compare two methods of assessing the slides (i.e. an exhaustive method and a rapid method). Two hundred eighty five post partum Holstein cows from 5 commercial dairy herds had a post partum evaluation between 21 and 47 days in milk (DIM). Sixty seven cows where diagnosed with clinical endometritis by transrectal and vaginoscopy examinations and were excluded from the study. Two hundred eighteen cows were enrolled for endometrial cytology and esterase activity test. The ovarian status was determined by transrectal examination. Computerized databank, dairy DSA (Dossier de Santé Animale) indexing all the cows was used to retrieve individual information for analysis. The percentage of neutrophils was significantly correlated with the LE from the uterus and cervix. The LE from cervix and uterus decreased significantly with DIM, however, they were not statistically associated with pregnancy risk at 90 DIM (n=186). Between 32-47 DIM, the percentage of neutrophils and risk of pregnancy at 90 DIM were associated (n=94, P=0.04). For the same period, survival analysis identified cows with > 2.6 % neutrophils on EC as subclinical endometritis cows with a prevalence of 56%. The two methods for assessing the slides were correlated by 81%. Subclinical endometritis diagnosed by endometrial cytology between 32 and 47 DIM was associated with reduced risk of pregnancy at first service.

No description available.

A modulação da resposta inflamatória uterina equina recebe atenção, devido a um distúrbio imunológico, que parece ocorrer em uma população de éguas, classificadas como suscetíveis à endometrite. O objetivo deste estudo foi verificar o efeito da corticoterapia aplicada na presença e na ausência de inflamação uterina sobre a proteômica e a concentração de óxido nítrico no líquido endometrial de éguas suscetíveis à endometrite. Éguas foram sincronizadas com 5 mg de prostaglandina F2α e após a verificação dos sinais de estro foram submetidas a quatro tratamentos. O primeiro foi o Controle, que não recebeu nenhum tipo de tratamento. O segundo foi o GC, onde as éguas receberam aplicação de um glicocorticóide, a cada 12 horas, por três dias consecutivos. O terceiro foi o Infectado, onde as éguas receberam uma infusão intrauterina de Streptococcus zoopepidemicus (1x109/mL) e o quarto foi o GC + Infectado, onde as éguas receberam a aplicação do glicocorticóide e a infusão intrauterina descrita acima. Doze horas após o final de cada tratamento, amostras de líquido endometrial puro e de lavados uterinos foram coletadas para análise proteômica e determinação de óxido nítrico, respectivamente. O primeiro artigo relata os dados preliminares da análise proteômica, com a contagem das bandas protéicas e a observação de 33 bandas protéicas no Controle, 54 no GC, 51 no Infectado e 72 no GC + Infectado. A corticoterapia pode induzir o aparecimento de um número maior de bandas protéicas, pois os géis com as maiores contagens foram nos tratamentos onde ela foi aplicada. No segundo artigo, foi realizada a identificação das bandas protéicas significativas, em relação à densidade óptica relativa e à frequência. A corticoterapia provocou uma alteração na proteômica do líquido endometrial, caracterizada pelo aumento e diminuição na densidade óptica relativa e na frequência de proteínas da fase aguda da inflamação, com as maiores alterações observadas quando a corticoterapia foi aplicada na presença do processo infeccioso. A infusão de Streptococcus zooepidemicus provocou alterações na proteômica do líquido endometrial, caracterizadas pelo aumento e diminuição na densidade óptica relativa e na frequência de proteínas da fase aguda da inflamação. Os resultados do estudo indicam que a corticoterapia provoca alterações imunológicas no endométrio equino, não apenas como depressiva, mas estimuladora da defesa local, através de uma ação imunomoduladora. No terceiro artigo, foi realizada a determinação da concentração de óxido nítrico, não sendo observada diferença significativa nos quatro tratamentos. Portanto, a corticoterapia provoca alterações proteômicas no líquido endometrial de éguas suscetíveis em estro, onde a presença de um estímulo inflamatório causado pela infusão intrauterina bacteriana induz a uma maior alteração, do que a ausência. A infecção do lúmen uterino provoca alterações proteômicas no líquido endometrial e a corticoterapia não influencia a concentração de óxido nítrico de éguas em estro. / The modulation of the equine uterine inflammatory response recieves much attention, due to an immunological disorder, which appears to happen in a population of mares, classified as suscetipble to endometritis. The aim of this study was to verify the effect of corticotherapy applied in the presence and in the absence of uterine inflammation on the proteomics and nitric oxide concentration of the endometrial fluid of mares susceptible to endometritis. Mares were synchronized with 5 mg prostaglandin F2α and after the observation of the signs of estrus were submitted to four treatments. The first was the Control, which did not recieve any kind of treatment. The second was the GC, where mares recieved the administration of a glucocorticoid, each 12 hours, for three consecutive days. The third was the Infected, where mares received an intrauterine infusion of Streptococcus zoopepidemicus (1x109/mL) and the fourth was the GC + Infected, where mares received the administration of glucocorticoid and intrauterine infusion as described above. Twelve hours after the end of each treatment, pure endometrial fluid and uterine flushings were collected for proteomic analysis and nitric oxide determination, respectively. The first article reports the preliminary data of the proteomic analysis, where protein band counts were done, being observed 33 protein bands in Control, 54 in GC, 51 in Infected and 72 in GC + Infected. Corticotherapy can induce the appearance of a higher number of protein bands, because the gels with the highest counts were in the treatments where it was applied. In the second article, the identification of the significative protein bands was done, regarding the relative optic density and frequency. Corticotherapy provoked an alteration in the endometrial proteomics, characterized by an increase and a decrease on the relative optic density and frequency of inflammatory acute phase proteins, with the major alterations observed when corticotherapy was applied in the presence of an infectious process. Streptococcus zooepidemicus infusion provoked alterations in the endometrial fluid proteomics, characterized by an increase and a decrease on the relative optic density and frequency of inflammatory acute phase proteins. Results from this study indicate that corticotherapy provokes immunological alterations in the equine endometrium, not only as depressor, but enhancer of local defense, through an immunomodulatory action. In the third article, the nitric oxide concentration was determined, with no significative diference observed in the four treatments. So, corticotherapy provokes alterations in the proteomics of the endometrial fluid of susceptible mares in estrus, where the presence of an inflammatory stimulus caused by intrauterine bacterial infusion induces a major alteration, than the absence. Uterine lumen infection provokes alterations in the proteomics of the endometrial fluid and corticotherapy does not influence nitric oxide concentration in mares in estrus.