Rotavírus (RV) é um importante patógeno viral que causa diarreia em leitões e indivíduos jovens de várias outras espécies animais. RV dos grupos A, B, C e E foram descritos como causa de diarreia em suínos. Este estudo reúne achados histopatológicos, imunohistoquímicos e de reação em cadeia da polimerase com transcriptase reversa (RT-PCR) presentes em quatro surtos de diarreia causados por RV de um e de múltiplos grupos na região sul do Brasil. Vacinação para RV não era aplicada em nenhuma das granjas estudadas. Necropsia, exames histológicos e imuni-histoquímicos foram realizados em 34 suínos de maternidade que apresentavam diarreia severa, além disso, realizou-se cultivo bacteriano e RT-PCR para RV dos grupos A, B e C, vírus da gastroenterite transmissível (TGEV), vírus da diarreia epidêmica dos suínos (PEDV), sapovírus (SaV), norovírus (NoV) e kobuvírus (Aichi vírus C) em 30 dessas amostras. Desidratação e conteúdo pastoso a líquido no cólon foram observados em todos os suínos. Exame histológico revelou atrofia de vilosidades em 29 casos, vacuolização de enterócitos em 27 casos e debris celulares na lâmina própria em 20 casos. Houve marcação imunohistoquímica positiva em 21 casos. RT-PCR foi positiva para RV em 20 casos e RV do grupo C foi o mais frequentemente detectado, presente em 17 amostras. Cultivo e isolamento de Escherichia coli ocorreu em todos os casos e quatro destes foram de E. coli α-hemolítica. Em 15 amostras houve isolamento de Clostridium sp. Sapovirus foi detectado em oito amostras, duas amostras foram positivas para norovírus e detectou-se kobuvírus em 11 animais. Os achados histológicos foram consistentes com infecção por RV e a imuno-histoquímica revelou dois padrões de marcação para o agente no intestino delgado. Os resultados de RT-PCR mostraram que o RV do grupo C foi o principal agente detectado neste estudo. O isolamento de E. coli e a detecção de SaV os destacou como agentes associados à infecção por RV. A detecção de kobuvírus o enfatiza como um novo candidato em associação com RV. / Rotavirus is an important viral pathogen causing diarrhea in piglets and other animal species worldwide. Groups A, B, C and E have been described causing diarrhea in swine. This study reunites histopathological, immunohistochemical and reverse transcriptase polymerase chain reaction (RT-PCR) findings present in four outbreaks of diarrhea caused by single and multiple groups of rotavirus in the south of Brazil. None of the herds studied applied vaccination against rotavirus. Necropsy, histological examination and immunohistochemistry were performed in 34 nursing piglets that presented severe diarrhea, bacterial culture and reverse transcriptase polymerase chain reaction (RT-PCR) for rotavirus from groups A, B and C, transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), sapovirus (SaV), norovirus (NoV) and kobuvirus (Aichi virus C) were carried out in 30 of the animals necropsied. Additionally, RT-PCR was performed in fecal pools from two outbreaks. Dehydration and fluid to pasty contents were observed in the colon of all 34 swine examined. Histological examination revealed villus atrophy in 29 cases, vacuolation of enterocytes in 27 cases and necrotic debris in the lamina propria of 20 cases. IHC was positive in 21 samples. RT-PCR was positive for rotavirus in 20 samples and group C rotavirus was the most frequently detected, present in 17 samples. Escherichia coli was isolated from all cases, and in four cases, it was α-hemolytic. Clostridium sp. was isolated from 15 samples. Sapovirus was detected in eight samples, samples from two animals were positive for norovirus and kobuvirus was detected from 11 samples. Histological findings were consistent with rotavirus infection and immunohistochemistry revealed two patterns of staining for rotavirus in the small intestine. RT-PCR results have shown group C rotavirus as the main agent detected in this study. The isolation of Escherichia coli and the detection of sapovirus highlighted them as possible agents associated to rotavirus infection. Kobuvirus detection has emphasized it as a new candidate in the association with rotavirus.

Destaca-se a Salmonella como um dos mais importantes patógenos veiculados por alimentos, devido ao fato de estar amplamente distribuído na natureza, possuir um grande número de reservatórios, apresentar sorotipos inespecíficos quanto aos hospedeiros e apresentar cepas multiresistentes aos antimicrobianos. Salmonella Enteritidis tornou-se um grande problema de saúde pública no Brasil, a partir da década de 80, quando foi associada a produtos avícolas. Desde 1962, a Salmonella Heidelberg tem sido isolada de aves e produtos derivados e aves no Brasil e tem reconhecida importância para saúde humana e em poedeiras comerciais na América do Norte. O presente trabalho teve por objetivo o estudo de diferentes cepas de Salmonella Heidelberg (SH) em comparação com Salmonella Enteritidis (SE) em pintinhos e frangos de corte no que diz respeito à capacidade de invasibilidade de órgãos, persistência de excreção fecal e alterações na morfometria intestinal. Para o estudo in vivo, 65 isolados de SH foram avaliados quanto à presença dos genes de virulência invA, lpfA e agfA e diferenciados por eletroforese em campo pulsado (PFGE). Outro objetivo foi testar uma mistura de ácidos orgânicos e óleos essenciais no controle de excreção fecal de SE. Duas amostras de SH não relacionadas pelo método de PFGE (SH23 e SH35) e uma cepa de SE padrão foram selecionadas para a inoculação das aves. As três cepas promoveram alteração na medida de vilosidades e criptas dos pintinhos, frente a grupo controle. A cepa SH35 promoveu maior reação inflamatória nos cecos. Ambas as cepas testadas foram capaz de atingir o fígado de pintinhos às 6 horas pós-inoculação. Tanto a SE como a SH demonstram persistência e intermitência de excreção fecal em frangos de corte até os 21dias pós-inoculação. A mistura de ácidos e óleos essenciais foi eficaz na redução da excreção fecal de SE, porém somente com o uso contínuo da mistura na ração. As cepas apresentaram comportamentos diferentes quando inoculadas em frangos de corte e devido à persistência destas nos intestinos dos frangos, prejuízo para as aves podem ocorrer, enquanto a subseqüente contaminação de carcaças, representa risco para a saúde humana. / Salmonella, one of the most important food borne pathogen due to the widely distribution in nature, large number of reservoirs, to show unspecific host serotypes and provide multiresistant strains to antibiotics. Salmonella Enteritidis became a big public health problem in Brasil, since 80’s, when it was correlated with poultry products. Since 1962, Salmonella Heidelberg has been isolated from birds and poultry products in Brazil, with importance in human health and layers in North America. The present work aimed to the study of different strains of Salmonella Heidelberg (SH) and compared with Salmonella Enteritidis (SE) in chicks and broiler chickens with regard to the ability of invasibilidade of organs, persistence of fecal excretion and changes in intestinal morphometry. For the in vivo study, 65 isolates of SH were evaluated for the presence of virulence genes invA, lpfA and agfA and differentiated by pulsed field gel electrophoresis (PFGE). Another objective was to test a organic acids and essential oils mixture for controlling SE fecal excretion. Two not related SH samples by the method of PFGE (SH23 and SH35) and a standard strain of SE were selected to the poultry inoculation. The three strains promoted changes in the measures of villi and crypts of the chicks intestine, when compared to the control group. The SH35strain promoted greater inflammatory reaction in the caeca. Both strains tested were able to achieve the chicks liver at 6 hours post inoculation. Both strains, SE and SH, showed persistence and intermittence of fecal excretion in broiler chickens until the 21days post-infection. The organic acids and essential oils mix was effective in reducing the SE fecal excretion, but only with the continuous use of the mix added to the feed. The strains showed different behavior when inoculated in broiler chickens, and because of the persistence of these strains in the chickens intestines, injury to birds may occur, while the subsequent carcasses contamination represents risk to human health.

Rotavírus (RV) é um importante patógeno viral que causa diarreia em leitões e indivíduos jovens de várias outras espécies animais. RV dos grupos A, B, C e E foram descritos como causa de diarreia em suínos. Este estudo reúne achados histopatológicos, imunohistoquímicos e de reação em cadeia da polimerase com transcriptase reversa (RT-PCR) presentes em quatro surtos de diarreia causados por RV de um e de múltiplos grupos na região sul do Brasil. Vacinação para RV não era aplicada em nenhuma das granjas estudadas. Necropsia, exames histológicos e imuni-histoquímicos foram realizados em 34 suínos de maternidade que apresentavam diarreia severa, além disso, realizou-se cultivo bacteriano e RT-PCR para RV dos grupos A, B e C, vírus da gastroenterite transmissível (TGEV), vírus da diarreia epidêmica dos suínos (PEDV), sapovírus (SaV), norovírus (NoV) e kobuvírus (Aichi vírus C) em 30 dessas amostras. Desidratação e conteúdo pastoso a líquido no cólon foram observados em todos os suínos. Exame histológico revelou atrofia de vilosidades em 29 casos, vacuolização de enterócitos em 27 casos e debris celulares na lâmina própria em 20 casos. Houve marcação imunohistoquímica positiva em 21 casos. RT-PCR foi positiva para RV em 20 casos e RV do grupo C foi o mais frequentemente detectado, presente em 17 amostras. Cultivo e isolamento de Escherichia coli ocorreu em todos os casos e quatro destes foram de E. coli α-hemolítica. Em 15 amostras houve isolamento de Clostridium sp. Sapovirus foi detectado em oito amostras, duas amostras foram positivas para norovírus e detectou-se kobuvírus em 11 animais. Os achados histológicos foram consistentes com infecção por RV e a imuno-histoquímica revelou dois padrões de marcação para o agente no intestino delgado. Os resultados de RT-PCR mostraram que o RV do grupo C foi o principal agente detectado neste estudo. O isolamento de E. coli e a detecção de SaV os destacou como agentes associados à infecção por RV. A detecção de kobuvírus o enfatiza como um novo candidato em associação com RV. / Rotavirus is an important viral pathogen causing diarrhea in piglets and other animal species worldwide. Groups A, B, C and E have been described causing diarrhea in swine. This study reunites histopathological, immunohistochemical and reverse transcriptase polymerase chain reaction (RT-PCR) findings present in four outbreaks of diarrhea caused by single and multiple groups of rotavirus in the south of Brazil. None of the herds studied applied vaccination against rotavirus. Necropsy, histological examination and immunohistochemistry were performed in 34 nursing piglets that presented severe diarrhea, bacterial culture and reverse transcriptase polymerase chain reaction (RT-PCR) for rotavirus from groups A, B and C, transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), sapovirus (SaV), norovirus (NoV) and kobuvirus (Aichi virus C) were carried out in 30 of the animals necropsied. Additionally, RT-PCR was performed in fecal pools from two outbreaks. Dehydration and fluid to pasty contents were observed in the colon of all 34 swine examined. Histological examination revealed villus atrophy in 29 cases, vacuolation of enterocytes in 27 cases and necrotic debris in the lamina propria of 20 cases. IHC was positive in 21 samples. RT-PCR was positive for rotavirus in 20 samples and group C rotavirus was the most frequently detected, present in 17 samples. Escherichia coli was isolated from all cases, and in four cases, it was α-hemolytic. Clostridium sp. was isolated from 15 samples. Sapovirus was detected in eight samples, samples from two animals were positive for norovirus and kobuvirus was detected from 11 samples. Histological findings were consistent with rotavirus infection and immunohistochemistry revealed two patterns of staining for rotavirus in the small intestine. RT-PCR results have shown group C rotavirus as the main agent detected in this study. The isolation of Escherichia coli and the detection of sapovirus highlighted them as possible agents associated to rotavirus infection. Kobuvirus detection has emphasized it as a new candidate in the association with rotavirus.

Made available in DSpace on 2015-03-26T13:54:52Z (GMT). No. of bitstreams: 1 texto completo.pdf: 435479 bytes, checksum: e3cdc5a0f6448d59185bed6203aceaff (MD5) Previous issue date: 2009-07-29 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / 126 commercial crossbred pigs weaned at 21 ± 1 days of age, with initial weight of 6.12 ± 0.70 kg, were allocated in randomized complete block designs. Two experiments were carried out to evaluate the effect of different levels of lactose and supplementation with glutamine and glutamic acid in pig diets on the performance, mucosa morphology and intestinal flora composition. Experiment 1 consisted of diets containing 0, 4, 8 and 12% lactose administered between 21 and 35 of age, conducted in six replicates with three animals per experimental unit. Diets in experiment 2 contained 0, 4 and 8% of lactose added with glutamine and glutamic acid, conducted in six replicates with three animals per experimental unit. Diets were lactose-free between days 36 and 49 in both experiments. Glutamine and glutamic acid supplementation from day 21 to 35 in animals from experiment 2 was maintained until the end of the experiment. Animal performance was evaluated between days 21 and 35 and between days 21 and 49. Intestinal mucosa morphology and intestinal flora composition were assayed at day 49. The ideal protein concept was used to prepare the isonitrogenous and isocaloric experimental mash diets, maintaining a constant relationship between lysine and other amino acids. Diets contained 2% spray-dried plasma and 0.05% tylosin growth promoter. No influence (P <0.05) was observed of the lactose levels on animal weight gain, feed intake and feed conversion between days 21 and 35 in experiment 1, and no residual effect of lactose (P <0.05) on the performance parameters, from day 21 to 49. Animals fed with diets containing 4, 8 or 12% lactose showed higher performance than animals fed with 0% lactose in absolute values, although there was no significant difference between the treatments. Villus height, crypt depth and the villus/crypt (small intestine) relationship were not affected by treatments. Augmented lactose levels altered the intestinal flora composition, increasing the number of Lactobacillus spp. in relation to total Eubacteria. Experiment 2 showed no effect of the interaction (P> 0.05) between lactose levels and glutamine and glutamic acid addition on the evaluated parameters, between 21 and 35 days and in the total experimental period. Lactose levels did not affect (P <0.05) animal performance in both periods. In absolute values, however, the treatment with 4% lactose had the best results of weight gain and feed intake in both periods. Addition of glutamine and glutamic acid in the diets affected (P <0.05) positive weight gain of piglets (days 21 to 49) and increased (P <0.05) villus height in the duodenum, jejunum and ileum of weaning piglets. Effect (P <0.05) of the addition of glutamine and glutamic acid on the villus/crypt relation was observed in the ileum. The experiments showed that lactose exerted a positive effect on the intestinal microbial ecology of pigs. The addition of glutamine and glutamic acid in the diets enhanced animal performance and intestinal villus height. / Com o objetivo de avaliar o efeito de diferentes níveis de lactose e da inclusão de glutamina + ácido glutâmico nas rações sobre o desempenho, a morfometria da mucosa e a composição da microbiota intestinal dos animais, utilizou-se 126 suínos mestiços comerciais, desmamados aos 21 ± 1 dias de idade, com peso inicial de 6,12 ± 0,70 Kg, distribuídos em delineamento de blocos ao acaso. Foram realizados dois experimentos. No experimento 1, os tratamentos consistiram em rações contendo 0, 4, 8 e 12% de lactose dos 21 aos 35 dias, com seis repetições e três animais por unidade experimental. No experimento 2 testou-se a inclusão de glutamina mais ácido glutâmico nas rações contendo 0, 4 e 8% de lactose, também com seis repetições e três animais por unidade experimental. Dos 36 aos 49 dias, nos dois experimentos, todas as rações foram isentas de lactose e os animais que receberam suplementação com glutamina mais ácido glutâmico dos 21 aos 35 dias no experimento 2 continuaram sendo suplementados até o final do experimento. Avaliou-se o desempenho dos animais dos 21 aos 35 e dos 21 aos 49 dias, a morfometria da mucosa intestinal e composição da microbiota intestinal aos 49 dias. As rações experimentais fareladas, isoprotéicas e isoenergéticas, foram formuladas observando-se o conceito de proteína ideal, mantendo-se constantes as relações aminoacídicas entre lisina e os demais aminoácidos essenciais. Todas as rações continham 2% de plasma sanguíneo "spray dried" em sua composição e 0,05% do antibiótico tilosina como promotor de crescimento. No Experimento 1, os níveis de lactose não influenciaram (P<0,05) o ganho de peso, o consumo de ração e a conversão alimentar dos leitões dos 21 aos 35 dias de idade. Também não foi observado efeito (P<0,05) residual da lactose sobre os parâmetros de desempenho, dos 21 aos 49 dias de idade. Apesar de não ter sido observado diferença significativa entre os tratamentos, observou-se que em valores absolutos os animais do tratamento com 0% de lactose apresentaram desempenho inferior ao dos animais alimentados com rações contendo 4, 8 ou 12% de lactose. Os tratamentos também não influenciaram (P<0,05) a altura das vilosidades, a profundidade das criptas e a relação vilo:cripta da mucosa do intestino delgado. O aumento do nível de lactose das rações alterou a composição da microbiota intestinal dos animais, promovendo aumento da quantidade de Lactobacillus spp. em relação ao total de Eubactéria. No experimento 2 não foi observada interação (P>0,05) entre o nível de lactose e a inclusão de glutamina + ácido glutâmico sobre nenhum dos parâmetros avaliados, tanto de 21 a 35 dias quanto no período total do experimento. Os níveis de lactose também não influenciaram (P<0,05) o desempenho dos leitões nos dois períodos avaliados. Entretanto, em valores absolutos, o tratamento com 4% de lactose apresentou os melhores resultados de ganho de peso e consumo de ração nos dois períodos. A inclusão de glutamina mais ácido glutâmico nas rações influenciou (P<0,05) positivamente o ganho de peso dos leitões dos 21 aos 49 dias de idade e promoveu aumento (P<0,05) da altura das vilosidades no duodeno, jejuno e íleo do intestino dos leitões. Na porção do intestino correspondente ao íleo, foi observado efeito (P<0,05) da inclusão de glutamina mais acido glutâmico nas dietas sobre a relação vilo:cripta dos leitões. Com base nos resultados obtidos nos dois experimentos, pode-se concluir que a lactose tem efeito positivo sobre a ecologia microbiana intestinal dos leitões e que a inclusão de glutamina mais acido glutâmico nas rações tem efeito benéfico sobre o desempenho e sobre a altura das vilosidades da mucosa intestinal dos animais.

Made available in DSpace on 2016-01-26T18:55:39Z (GMT). No. of bitstreams: 1 Gabriel Sakita.pdf: 2676800 bytes, checksum: 1441b473ee5d17d030119d3eef69a692 (MD5) Previous issue date: 2014-03-28 / Functional and nutraceuticals foods are defined as those which by virtue of physiologically active components, promote health benefits. Among these foods are prebiotics. Prebiotics are defined as non-digestible food compounds that beneficially affect the host by selectively stimulating the growth and/or activity of populations of desirable bacteria in the colon, inhibiting the multiplication of pathogens and ensuring benefits to host health. Currently, these compounds have been used as an alternative to growth promoters in order to maintain the beneficial balance of intestinal microbiota, especially in young animals or in imminent stress condition. The objective of the research project was to evaluate the use of three different types of prebiotics, Bio-MosTM, ActigenTM e ImmunowallTM, based MOS, which are sold in the market, on cumulative feed intake, percentage of weight gain, feed conversion ratio, feed efficiency, villus height and area, crypt depth, and goblet cell counting of albino rats (Rattus norvegicus) Wistar. The study was conducted to compare the product which has greater benefits to the animal organism. / Alimentos funcionais ou nutracêuticos são definidos como aqueles que em virtude de componentes fisiologicamente ativos, promovem benefícios para a saúde. Dentre esses alimentos encontram-se os prebióticos. Os prebióticos são definidos como compostos alimentares não digeríveis que afetam beneficamente o hospedeiro por estimularem seletivamente a proliferação e/ou atividade de populações de bactérias desejáveis no cólon, inibindo a multiplicação de patógenos e garantindo benefícios a saúde do hospedeiro. Atualmente, estes compostos vêm sendo utilizados como alternativa aos promotores de crescimento com o objetivo de manter o equilíbrio benéfico da microbiota intestinal, especialmente em animais jovens ou em iminente condição de estresse. O objetivo do projeto de pesquisa foi avaliar a utilização de três diferentes tipos de prebióticos, Bio-MosTM, ActigenTM e ImmunowallTM, a base de mananoligossacarídeos, que são comercializados no mercado, sobre os parâmetros de consumo acumulado de ração, percentual de ganho de peso, índice conversão alimentar, coeficiente de eficiência alimenta, altura e área das vilosidades, profundidade das criptas e contagem de células caliciformes duodenais de ratos albinos (Rattus novergicus) linhagem Wistar. O estudo foi realizado para comparar qual produto apresenta maiores benefícios sobre o organismo animal.

Made available in DSpace on 2016-07-18T17:53:12Z (GMT). No. of bitstreams: 1 Gabriel Sakita.pdf: 2676800 bytes, checksum: 1441b473ee5d17d030119d3eef69a692 (MD5) Previous issue date: 2014-03-28 / Functional and nutraceuticals foods are defined as those which by virtue of physiologically active components, promote health benefits. Among these foods are prebiotics. Prebiotics are defined as non-digestible food compounds that beneficially affect the host by selectively stimulating the growth and/or activity of populations of desirable bacteria in the colon, inhibiting the multiplication of pathogens and ensuring benefits to host health. Currently, these compounds have been used as an alternative to growth promoters in order to maintain the beneficial balance of intestinal microbiota, especially in young animals or in imminent stress condition. The objective of the research project was to evaluate the use of three different types of prebiotics, Bio-MosTM, ActigenTM e ImmunowallTM, based MOS, which are sold in the market, on cumulative feed intake, percentage of weight gain, feed conversion ratio, feed efficiency, villus height and area, crypt depth, and goblet cell counting of albino rats (Rattus norvegicus) Wistar. The study was conducted to compare the product which has greater benefits to the animal organism. / Alimentos funcionais ou nutracêuticos são definidos como aqueles que em virtude de componentes fisiologicamente ativos, promovem benefícios para a saúde. Dentre esses alimentos encontram-se os prebióticos. Os prebióticos são definidos como compostos alimentares não digeríveis que afetam beneficamente o hospedeiro por estimularem seletivamente a proliferação e/ou atividade de populações de bactérias desejáveis no cólon, inibindo a multiplicação de patógenos e garantindo benefícios a saúde do hospedeiro. Atualmente, estes compostos vêm sendo utilizados como alternativa aos promotores de crescimento com o objetivo de manter o equilíbrio benéfico da microbiota intestinal, especialmente em animais jovens ou em iminente condição de estresse. O objetivo do projeto de pesquisa foi avaliar a utilização de três diferentes tipos de prebióticos, Bio-MosTM, ActigenTM e ImmunowallTM, a base de mananoligossacarídeos, que são comercializados no mercado, sobre os parâmetros de consumo acumulado de ração, percentual de ganho de peso, índice conversão alimentar, coeficiente de eficiência alimenta, altura e área das vilosidades, profundidade das criptas e contagem de células caliciformes duodenais de ratos albinos (Rattus novergicus) linhagem Wistar. O estudo foi realizado para comparar qual produto apresenta maiores benefícios sobre o organismo animal.

Prenatal diagnostics is headed in several directions - towards visualization of fetuses and biochemical, cytogenetic and molecular genetic diagnostics in laboratories. Whereas visualization of fetuses does not a priori represent any direct risk for pregnancy and does not increase the number of potential pregnancy complications, this is not always the case with the laboratory testing. There are known risks connected with invasive methods of prenatal diagnostics. The number of potential unintentional pregnancy complications and losses as well as the technical and economic aspects of invasive prenatal diagnostics lead to attempts of identifying ways of detecting any potentially affected individuals by screening methods, thus minimizing the undesirable impact of invasive diagnostics on the pregnant population. The more precise the selective criteria, the lesser the number of pregnant women exposed to invasive exams. Another way of decreasing the number of unintentional complications in relation to invasive diagnostics is to simplify and improve the fetal samples harvesting methods during pregnancy. The work primarily focused on two areas: Determination of the relation between fraction shortening of the left and right ventricles and a fetal chromosomal complement, and verification of reliability of a new method...

Prenatal diagnostics is headed in several directions - towards visualization of fetuses and biochemical, cytogenetic and molecular genetic diagnostics in laboratories. Whereas visualization of fetuses does not a priori represent any direct risk for pregnancy and does not increase the number of potential pregnancy complications, this is not always the case with the laboratory testing. There are known risks connected with invasive methods of prenatal diagnostics. The number of potential unintentional pregnancy complications and losses as well as the technical and economic aspects of invasive prenatal diagnostics lead to attempts of identifying ways of detecting any potentially affected individuals by screening methods, thus minimizing the undesirable impact of invasive diagnostics on the pregnant population. The more precise the selective criteria, the lesser the number of pregnant women exposed to invasive exams. Another way of decreasing the number of unintentional complications in relation to invasive diagnostics is to simplify and improve the fetal samples harvesting methods during pregnancy. The work primarily focused on two areas: Determination of the relation between fraction shortening of the left and right ventricles and a fetal chromosomal complement, and verification of reliability of a new method...

Crohns sjukdom är en kronisk inflammatorisk tarmsjukdom av okänd orsak. Det tidigaste tecknet på Crohns sjukdom är mikroskopiska sår i det s.k. follikelassocierade epitelet (FAE) som täcker ansamlingar av immunceller i tarmen. FAE är specialiserat för att fånga innehåll från tarmen och transportera det till underliggande immunvävnad. Denna funktion är viktig för att inducera skyddande immunsvar, men den utgör också en ingångsväg för sjukdomsalstrande bakterier. Crohns sjukdom är associerat med ett kraftigt ökat immunsvar mot bakterier, och sjukdomsförloppet kan ändras av stress. Det övergripande syftet med avhandlingen var att studera effekterna av stress på FAE samt att undersöka rollen av FAE vid utvecklingen av tarminflammation, särskilt vid Crohns sjukdom. Inledningsvis studerades effekterna av psykologisk stress på FAE. Stressade råttor uppvisade ökad genomsläpplighet av bakterier efter stress, och passagen var högre i FAE än i vanligt epitel. Efterföljande experiment visade att stressförändringarna i slemhinnan regleras via kortikotropinfrisättande hormon och mastceller. Vidare visade det sig att vasoaktiv intestinal peptid kunde efterlikna stressens effekter på genomsläppligheten, och att detta kunde förhindras genom att blockera mastcellerna. Studier av tunntarmsslemhinna från patienter med icke-inflammatorisk tarmsjukdom och friska kontroller visade en högre passage av bakterier i FAE än i vanligt epitel. Hos patienter med Crohns sjukdom var bakteriepassagen genom FAE betydligt ökad jämfört med kontroller. Resultaten från detta avhandlingsarbete visar att stress kan förändra upptaget av bakterier från tarmen via FAE, med mekanismer som innefattar kortikotropinfrisättande hormon och mastceller. Detta har gett nya kunskaper kring regleringen av slemhinnebarriären. Vidare presenterar denna avhandling nya insikter i sjukdomsuppkomsten vid Crohns sjukdom genom att påvisa en tidigare okänd defekt i barriärfunktionen i FAE. / The earliest observable signs of Crohn’s disease are microscopic erosions in the follicle-associated epithelium (FAE) covering the Peyer’s patches. The FAE, which contains M cells, is specialised in sampling of luminal content and delivery to underlying immune cells. This sampling is crucial for induction of protective immune responses, but it also provides a route of entry for microorganisms into the mucosa. Crohn’s disease is associated with an increased immune response to bacteria, and the disease course can be altered by stress. The overall aim of this thesis was to study the effects of stress on the FAE and elucidate the role of FAE in the development of intestinal inflammation, specifically Crohn’s disease. Initially, rats were submitted to acute and chronic water avoidance stress to study the effects of psychological stress on the FAE. Stressed rats showed enhanced antigen and bacterial passage, and the passage was higher in FAE than in regular villus epithelium (VE). Further, stress gave rise to ultrastructural changes. Subsequent experiments revealed the stress-induced increase in permeability to be regulated by corticotropin-releasing hormone and mast cells. Furthermore, vasoactive intestinal peptide (VIP) mimicked the stress effects on permeability, and the VIP effects were inhibited by a mast cell stabiliser. Human studies of ileal mucosa from patients with non-inflammatory disease and healthy controls showed a higher antigen and bacterial passage in FAE than in VE. In patients with Crohn’s disease, the bacterial passage across the FAE was significantly increased compared to non-inflammatory and inflammatory controls (ulcerative colitis). Furthermore, there was an enhanced uptake of bacteria into dendritic cells, and augmented TNF-α release in Crohn’s disease mucosa. Taken together this thesis shows that stress can modulate the uptake of luminal antigens and bacteria via the FAE, through mechanisms involving CRH and mast cells. It further shows that human ileal FAE is functionally distinct from VE, and that Crohn’s disease patients exhibit enhanced FAE permeability compared to inflammatory and non-inflammatory controls. This thesis presents novel insights into regulation of the FAE barrier, as well as into the pathophysiology of Crohn’s disease by demonstrating a previously unrecognised defect of the FAE barrier function in ileal Crohn’s disease.

In this thesis, we deal with the issue of invasive methods of prenatal care from a nursing perspective using standardized classification schemes NANDA, NIC and NOC, which offer many opportunities to streamline, simplify and improve the care of women who undergo invasive procedures. The theoretical part is focused on screening in prenatal care, individual invasive methods and the most common congenital malformations of the child. Furthermore, standardized classification systems NANDA, NIC and NOC, which represent a common nursing language. The empirical part of the dissertation is processed using qualitative strategy. To collect data, we selected the following research methods: content analysis of documents, evaluation using Fehring's model. The first goal was to detect and identify the problems that women face after amniocentesis (AMC), chorionic villus sampling (CVS) and cordocentesis and second goal was to assess the applicability of selected issues of classification systems NANDA, NIC and NOC with Fehring's model of validation of nursing diagnoses. Using content analysis, we have selected three nursing diagnosis from the NANDA International Taxonomy II, from publication of Nursing Interventions clasification (NIC) 3 nursing interventions and from Nursing Outcomes Clasification (NOC) 4 expected results, which are focused on the respondents after invasive methods. From selected nursing diagnoses of NANDA, NIC interventions and NOC expected results, we have developed a research form, specific questionnaire we submitted to the validation by respondents. The survey was carried out from January to March 2015 in the polyclinic GENNET Ltd. - Centrum of Medical genetics and reproductive medicine and in a private clinic OG Medical Center Ltd. The sample consisted of 34 intentionally selected respondents who underwent amniocentesis, chorionic villus sampling, or cordocentesis. The outcome of this thesis is the basic concept of the nursing diagnosis, according to NANDA-I, NIC and NOC aimed at women after invasive methods for prenatal care. Research has shown that with the the vast majority of respondents significantly exhibited symptoms of fear and anxiety. In contrast, in terms of the respondents reported that the knowledge of the issue is sufficient. This result is influenced by the fact that respondents find a lot of information on the Internet, but that may not always be true. Unfortunately, the results showed that it is more a problem of communication and interest of the medical staff.