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Correlates of protection among HIV-1-infected African childrenChakraborty, Rana January 2003 (has links)
No description available.
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Influência da vitamina e na profilaxia e tratamento de broncopneumonia em bezerrosRocha, Ana Elisa Arruda [UNESP] 31 August 2006 (has links) (PDF)
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rocha_aea_me_botfmvz.pdf: 607611 bytes, checksum: cfec96393ada5ad9ec693eef7143fc85 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Estudou-se a broncopneumonia de ocorrência natural em bezerros mantidos em bezerreiro coletivo, e a influência da administração intramuscular de acetato de DL-a-tocoferol na profilaxia e tratamento desta enfermidade. Foram avaliados 60 bezerros machos, com idade máxima de 10 dias, sorteados para comporem os grupos experimentais: GSV (grupo sem vitamina) e GCV (grupo com vitamina). A suplementação foi precedida por exame físico, colheita de sangue para hemograma, determinação de proteínas séricas e GGT, e lavado traqueobrônquico para exame citológico (D0). Os animais de ambos os grupos foram conduzidos ao bezerreiro coletivo, onde permaneceram até o 21º dia (D0 ao D21). Foram distribuídos aleatoriamente, 2 a 2, em baias individuais de 1,20m de largura por 2 m de comprimento. Independentemente do grupo experimental, os animais foram avaliados por exame físico diário e, na presença de sinais clínicos indicativos de broncopneumonia (DX), foram retirados do bezerreiro, avaliados por hemograma e citologia do lavado traqueobrônquico, e tratados com enrofloxacina. Durante o tratamento, o exame físico foi realizado diariamente, e os exames complementares, hemograma e citologia do lavado traqueobrônquico, repetidos uma semana após seu término (DY). Nos animais sadios, além do exame físico diário e dos exames complementares já realizados no D0, foram repetidos hemograma e citologia do lavado traqueobrônquico no último dia (D21) do experimento. Comparou-se a ocorrência de broncopneumonia e a evolução do tratamento nos dois grupos estudados. A suplementação com vitamina E não teve influência na profilaxia e no tratamento de broncopneumonia de bezerros. / Bronchopneumonia of natural occurrence in calf housed calves was studied, and the influence of intramuscular administration of DL-a-tocoferol acetate on prophylaxis and treatment of this disease. Sixty male calves, with maximum age of 10 days, were evaluated, draftees to compose the experimental groups: GSV (animals without supplementation) and GCV (animals with supplementation). Supplementation was preceded by physical examination, collection of blood samples for hemogram, total protein determination and GGT, and tracheobronchial wash for cytological examination (D0). The animals were taken to the calf housing, where they remained until 21º day (D0 to D21). They were distributed, 2 - 2, in individual pens of 1,20m of width for 2 m of length. Independently of the group, all animals were evaluated by physical examination daily and, in presence of indicative clinical signs of bronchopneumonia (DX), they were removed out of the calf housing, evaluated for hemogram and cytology of tracheobronchial wash, and treated with enrofloxacin. During the treatment, physical examination was carried through daily and the complementary examinations, hemogram and cytology of the tracheobronchial wash were repeated one week after its ending (DY). In healthy animals, beyond the daily physical examination and complementary examinations already done on D0, hemogram and cytology of tracheobronchial wash were repeated in the last day of the experiment (D21). Occurrence of bronchopneumonia and evolution of the treatment in both studied groups were compared. The supplementation with vitamin E did not have influence on bronchopneumonia prophylaxis or treatment. Key words: calves, bronchopneumonia, vitamin E, tracheobronchial wash, prophylaxis.
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Influência da vitamina e na profilaxia e tratamento de broncopneumonia em bezerros /Rocha, Ana Elisa Arruda. January 2006 (has links)
Orientador: Roberto Calderon Gonçalves / Banca: Simone Biagio Chiacchio / Banca: Laurenil Gaste / Resumo: Estudou-se a broncopneumonia de ocorrência natural em bezerros mantidos em bezerreiro coletivo, e a influência da administração intramuscular de acetato de DL-a-tocoferol na profilaxia e tratamento desta enfermidade. Foram avaliados 60 bezerros machos, com idade máxima de 10 dias, sorteados para comporem os grupos experimentais: GSV (grupo sem vitamina) e GCV (grupo com vitamina). A suplementação foi precedida por exame físico, colheita de sangue para hemograma, determinação de proteínas séricas e GGT, e lavado traqueobrônquico para exame citológico (D0). Os animais de ambos os grupos foram conduzidos ao bezerreiro coletivo, onde permaneceram até o 21º dia (D0 ao D21). Foram distribuídos aleatoriamente, 2 a 2, em baias individuais de 1,20m de largura por 2 m de comprimento. Independentemente do grupo experimental, os animais foram avaliados por exame físico diário e, na presença de sinais clínicos indicativos de broncopneumonia (DX), foram retirados do bezerreiro, avaliados por hemograma e citologia do lavado traqueobrônquico, e tratados com enrofloxacina. Durante o tratamento, o exame físico foi realizado diariamente, e os exames complementares, hemograma e citologia do lavado traqueobrônquico, repetidos uma semana após seu término (DY). Nos animais sadios, além do exame físico diário e dos exames complementares já realizados no D0, foram repetidos hemograma e citologia do lavado traqueobrônquico no último dia (D21) do experimento. Comparou-se a ocorrência de broncopneumonia e a evolução do tratamento nos dois grupos estudados. A suplementação com vitamina E não teve influência na profilaxia e no tratamento de broncopneumonia de bezerros. / Abstract: Bronchopneumonia of natural occurrence in calf housed calves was studied, and the influence of intramuscular administration of DL-a-tocoferol acetate on prophylaxis and treatment of this disease. Sixty male calves, with maximum age of 10 days, were evaluated, draftees to compose the experimental groups: GSV (animals without supplementation) and GCV (animals with supplementation). Supplementation was preceded by physical examination, collection of blood samples for hemogram, total protein determination and GGT, and tracheobronchial wash for cytological examination (D0). The animals were taken to the calf housing, where they remained until 21º day (D0 to D21). They were distributed, 2 - 2, in individual pens of 1,20m of width for 2 m of length. Independently of the group, all animals were evaluated by physical examination daily and, in presence of indicative clinical signs of bronchopneumonia (DX), they were removed out of the calf housing, evaluated for hemogram and cytology of tracheobronchial wash, and treated with enrofloxacin. During the treatment, physical examination was carried through daily and the complementary examinations, hemogram and cytology of the tracheobronchial wash were repeated one week after its ending (DY). In healthy animals, beyond the daily physical examination and complementary examinations already done on D0, hemogram and cytology of tracheobronchial wash were repeated in the last day of the experiment (D21). Occurrence of bronchopneumonia and evolution of the treatment in both studied groups were compared. The supplementation with vitamin E did not have influence on bronchopneumonia prophylaxis or treatment. Key words: calves, bronchopneumonia, vitamin E, tracheobronchial wash, prophylaxis. / Mestre
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Aspectos clínicos, hematológicos e imunológicos em fêmeas Holandesas persistentemente infectadas pelo Vírus da Diarreia Viral Bovina (BVDV) / Clinical, hematological and imunological findings in persistently infected Holstein heifers and cows by Bovine Viral Diarrhea VirusSobreira, Natália Meirelles 23 February 2018 (has links)
O objetivo deste estudo foi avaliar os aspectos clínicos, a eficiência produtiva, reprodutiva, a resposta imune inata e humoral em fêmeas da raça Holandesa persistentemente infectadas (PI) com BVDV. Para tanto, foram selecionadas 25 PI´s por meio do ELISA antígeno, das quais oito eram bezerras ≤ 12 meses; seis novilhas entre 13 e 24 meses; e onze animais de 25 a 36 meses de idade. O grupo não infectado (NI) foi composto por fêmeas negativas com faixas etárias pareadas às PIs. Em geral, as fêmeas PIs apresentaram maior frequência de diarreia (P=0,012) e Doença Respiratória Bovina (DRB) (P=0,004), especialmente àquelas entre 25 a 36 meses de idade. As PIs apresentaram maior concentração sérica de haptoglobina (P=0,017). As vacas NI´s produziram mais leite (8-12 litros) do que as PIs (P≤0.011). A contagem de células somáticas (CCS) foi maior para as vacas PI´s em relação NI´s durante a lactação (P≤0,066). A média de CCS observada foi de 0,2-0,5 x 105 (células/mL) e 2,0-10,5 x 105 (células/mL) nos grupos PI e NI, respectivamente. A idade na primeira inseminação (P=0,001) e o número de inseminação necessário para a primeira prenhez foi maior para PI do que nas novilhas NI´s (P=0,051). Na avaliação hematológica, as PI´s apresentaram menores valores de hemoglobina (P=0,098), hematócrito (P=0,084), hemoglobina corpuscular média (P=0,092), plaquetas (P=0,040), plaquetócrito (P=0,059) e linfócitos (número absoluto P=0,075; relativo P=0,092) em relação às NI´s. Em contraste, a porcentagem de monócitos foi maior em PI do que animais NI (P=0,014). Em geral, a proporção (%) e a intensidade da fagocitose (MFI) para Staphylococcus aureus e Escherichia coli foram maiores nas PIs do que animais NI (P<0,079). A porcentagem de células que produziram espécies reativas de oxigênio (EROs) endógeno foi menor em PI do que animais de NI (P=0,005). Padrão semelhante foi observado na porcentagem de células que produziram EROs após estímulo com Staphylococcus aureus (P=0,000) ou Escherichia coli (P=0,000). Em contraste, a intensidade da produção de EROS basal foi maior nas PI´s do que nas NI´s (P=0,011). Um padrão semelhante foi observado para a intensidade da produção de EROs quando as células foram estimuladas com Staphylococcus aureus (P=0,011) ou Escherichia coli (P= 0,013). Por meio da soroneutralização, observou-se que os títulos médios dos anticorpos neutralizantes para as BVDV, BRSV e BoHV foram menor no grupo PI (P=0,000). No entanto, os títulos de anticorpos para BPIV-3 foram semelhantes entre os grupos (P=0,146). A proteína sérica total (PT) foi menor nas PI em relação as NI´s (P=0,021). As fêmeas PIs apresentaram maior frequência de diarreia e DRB, além da menor produção leiteira, diminuição da qualidade do leite e menor fertilidade. O leucograma demonstrou diminuição dos tipos celulares, exceto monócitos. No geral, houve resposta imune inata foi exacerbada nas fêmeas PIs, provavelmente pela menor eficiência na produção de EROs. Em relação ao sistema imune específico observou-se linfopenia e títulos nulos ou baixos de anticorpos neutralizantes contra as viroses respiratórias. / The aim of this study was to evaluate the clinical problems, the efficiency of production, reproduction, the innate and humoral immune response of Holstein heifers and cows persistently infected (PI) with BVDV. Therefore, 25 PI animals were selected using ELISA antigen of which, eight were heifers ≤ 12 months; six were heifers between 13 to 24 months; and eleven were animals from 25 to 36 months old. An uninfected group (NI) composed by negative females with same age of PI´s. We observed that PI´s presented more frequency of diarrhea (P=0.012) and Bovine Respiratory Disease (BRD) score (P=0.004), especially the animals from 25 to 36 months old. The PI´s showed higher concentration of haptoglobin (P=0.017). During the lactations the milk production from NI cows was higher (8 to 18,2 litters) than PI (P≤0.011). Somatic cells count (SCC) were higher for PI than NI cows across milk production (P≤0.066). The mean of SCC observed was 0,2-0,5 x105 (cells/mL) in NI group and 2,0-10,5 x105 (cells/mL) for PI cows. The age at first insemination (P=0.001) and number of insemination required for the first pregnancy was higher for PI than NI heifers (P=0.051). For hematological evaluation, PI´s showed lower hemoglobin (P=0,098), hematocrit (P=0,084), mean corpuscular hemoglobin (P=0,092), platelet (P=0,040), plateletcrit (P=0,059) and lymphocytes (absolute number P=0.092; relative P=0,075) than NI. In contrast, the percentage of monocytes was higher in PI than NI animals (P=0.014). In aggregate, the percent of phagocytic cells and the average phagocytic avidity (expressed as mean fluorescence intensity, MFI) for Staphylococcus aureus and Escherichia coli were higher in PI than NI animals (P≤ 0.079). The percentage of cells producing endogenous reactive oxygen species (ROS) activity was lower in PI than NI animals (P=0.005). A similar pattern was observed in the percentage of cells producing ROS after exposure to Staphylococcus aureus (P= 0.000) or Escherichia coli (P=0.000). In contrast, the value for endogenous ROS per cell activity (MFI) was higher in PI than NI animals (P=0.011). A similar pattern was observed for ROS MFI after stimulating with Staphylococcus aureus (P=0.011) or Escherichia coli stimulation (P=0.013). The median conventional SN titers for specific antibody recognizing BVDV, BRSV and BoHV-1 in serum were lower in PI than NI animals (P=0.000). The antibody titers for BPIV3 were similar between the groups animals (P=0,146). In aggregate, the total serum protein (TP) was lower in PI than NI cattle (P=0.021). In summary, PI heifers had more diarrhea, BRD, produced less milk, worst milk quality and poorer reproductive performance than NI cattle. Leukogram showed decreased cell types except monocytes. Overall, there was a higher endogenous level innate cell reactivity in PI heifers and cows than NI animals, probably because of the lower efficiency in ROS production. In relation to the specific immune system it appear that the PI animals were less efficient in clearance of antigens due to lower numbers of lymphocytes and related lower titers of specific antibody against respiratory viruses.
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Aspectos clínicos, hematológicos e imunológicos em fêmeas Holandesas persistentemente infectadas pelo Vírus da Diarreia Viral Bovina (BVDV) / Clinical, hematological and imunological findings in persistently infected Holstein heifers and cows by Bovine Viral Diarrhea VirusNatália Meirelles Sobreira 23 February 2018 (has links)
O objetivo deste estudo foi avaliar os aspectos clínicos, a eficiência produtiva, reprodutiva, a resposta imune inata e humoral em fêmeas da raça Holandesa persistentemente infectadas (PI) com BVDV. Para tanto, foram selecionadas 25 PI´s por meio do ELISA antígeno, das quais oito eram bezerras ≤ 12 meses; seis novilhas entre 13 e 24 meses; e onze animais de 25 a 36 meses de idade. O grupo não infectado (NI) foi composto por fêmeas negativas com faixas etárias pareadas às PIs. Em geral, as fêmeas PIs apresentaram maior frequência de diarreia (P=0,012) e Doença Respiratória Bovina (DRB) (P=0,004), especialmente àquelas entre 25 a 36 meses de idade. As PIs apresentaram maior concentração sérica de haptoglobina (P=0,017). As vacas NI´s produziram mais leite (8-12 litros) do que as PIs (P≤0.011). A contagem de células somáticas (CCS) foi maior para as vacas PI´s em relação NI´s durante a lactação (P≤0,066). A média de CCS observada foi de 0,2-0,5 x 105 (células/mL) e 2,0-10,5 x 105 (células/mL) nos grupos PI e NI, respectivamente. A idade na primeira inseminação (P=0,001) e o número de inseminação necessário para a primeira prenhez foi maior para PI do que nas novilhas NI´s (P=0,051). Na avaliação hematológica, as PI´s apresentaram menores valores de hemoglobina (P=0,098), hematócrito (P=0,084), hemoglobina corpuscular média (P=0,092), plaquetas (P=0,040), plaquetócrito (P=0,059) e linfócitos (número absoluto P=0,075; relativo P=0,092) em relação às NI´s. Em contraste, a porcentagem de monócitos foi maior em PI do que animais NI (P=0,014). Em geral, a proporção (%) e a intensidade da fagocitose (MFI) para Staphylococcus aureus e Escherichia coli foram maiores nas PIs do que animais NI (P<0,079). A porcentagem de células que produziram espécies reativas de oxigênio (EROs) endógeno foi menor em PI do que animais de NI (P=0,005). Padrão semelhante foi observado na porcentagem de células que produziram EROs após estímulo com Staphylococcus aureus (P=0,000) ou Escherichia coli (P=0,000). Em contraste, a intensidade da produção de EROS basal foi maior nas PI´s do que nas NI´s (P=0,011). Um padrão semelhante foi observado para a intensidade da produção de EROs quando as células foram estimuladas com Staphylococcus aureus (P=0,011) ou Escherichia coli (P= 0,013). Por meio da soroneutralização, observou-se que os títulos médios dos anticorpos neutralizantes para as BVDV, BRSV e BoHV foram menor no grupo PI (P=0,000). No entanto, os títulos de anticorpos para BPIV-3 foram semelhantes entre os grupos (P=0,146). A proteína sérica total (PT) foi menor nas PI em relação as NI´s (P=0,021). As fêmeas PIs apresentaram maior frequência de diarreia e DRB, além da menor produção leiteira, diminuição da qualidade do leite e menor fertilidade. O leucograma demonstrou diminuição dos tipos celulares, exceto monócitos. No geral, houve resposta imune inata foi exacerbada nas fêmeas PIs, provavelmente pela menor eficiência na produção de EROs. Em relação ao sistema imune específico observou-se linfopenia e títulos nulos ou baixos de anticorpos neutralizantes contra as viroses respiratórias. / The aim of this study was to evaluate the clinical problems, the efficiency of production, reproduction, the innate and humoral immune response of Holstein heifers and cows persistently infected (PI) with BVDV. Therefore, 25 PI animals were selected using ELISA antigen of which, eight were heifers ≤ 12 months; six were heifers between 13 to 24 months; and eleven were animals from 25 to 36 months old. An uninfected group (NI) composed by negative females with same age of PI´s. We observed that PI´s presented more frequency of diarrhea (P=0.012) and Bovine Respiratory Disease (BRD) score (P=0.004), especially the animals from 25 to 36 months old. The PI´s showed higher concentration of haptoglobin (P=0.017). During the lactations the milk production from NI cows was higher (8 to 18,2 litters) than PI (P≤0.011). Somatic cells count (SCC) were higher for PI than NI cows across milk production (P≤0.066). The mean of SCC observed was 0,2-0,5 x105 (cells/mL) in NI group and 2,0-10,5 x105 (cells/mL) for PI cows. The age at first insemination (P=0.001) and number of insemination required for the first pregnancy was higher for PI than NI heifers (P=0.051). For hematological evaluation, PI´s showed lower hemoglobin (P=0,098), hematocrit (P=0,084), mean corpuscular hemoglobin (P=0,092), platelet (P=0,040), plateletcrit (P=0,059) and lymphocytes (absolute number P=0.092; relative P=0,075) than NI. In contrast, the percentage of monocytes was higher in PI than NI animals (P=0.014). In aggregate, the percent of phagocytic cells and the average phagocytic avidity (expressed as mean fluorescence intensity, MFI) for Staphylococcus aureus and Escherichia coli were higher in PI than NI animals (P≤ 0.079). The percentage of cells producing endogenous reactive oxygen species (ROS) activity was lower in PI than NI animals (P=0.005). A similar pattern was observed in the percentage of cells producing ROS after exposure to Staphylococcus aureus (P= 0.000) or Escherichia coli (P=0.000). In contrast, the value for endogenous ROS per cell activity (MFI) was higher in PI than NI animals (P=0.011). A similar pattern was observed for ROS MFI after stimulating with Staphylococcus aureus (P=0.011) or Escherichia coli stimulation (P=0.013). The median conventional SN titers for specific antibody recognizing BVDV, BRSV and BoHV-1 in serum were lower in PI than NI animals (P=0.000). The antibody titers for BPIV3 were similar between the groups animals (P=0,146). In aggregate, the total serum protein (TP) was lower in PI than NI cattle (P=0.021). In summary, PI heifers had more diarrhea, BRD, produced less milk, worst milk quality and poorer reproductive performance than NI cattle. Leukogram showed decreased cell types except monocytes. Overall, there was a higher endogenous level innate cell reactivity in PI heifers and cows than NI animals, probably because of the lower efficiency in ROS production. In relation to the specific immune system it appear that the PI animals were less efficient in clearance of antigens due to lower numbers of lymphocytes and related lower titers of specific antibody against respiratory viruses.
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Avaliação clínica e estudo da ocorrência de broncopneumonia em bovinos / Clinical evaluation and study of the occurence of bronchopneumonia in cattleOliveira, Bernardo Augusto França Dias de 27 July 2015 (has links)
As doenças respiratórias são consideradas um sério problema de sanidade, causando grandes perdas econômicas devido aos altos índices de morbidade e mortalidade, sendo responsáveis por 10 a 40% das mortes em animais adultos e por 17% das mortes durante o período perinatal. O objetivo deste trabalho foi determinar a ocorrência de agentes e buscar associações entre agentes e a broncopneumonia e a associação de sinais clínicos com os agentes presentes. Foram avaliados clinicamente e os dados coletados em ficha própria 96 animais de dois rebanhos da região de Bragança Paulista e Mococa ambas no Estado de São Paulo. Sangue total e lavado traqueobrônquico instilado em haste flexível e o restante da alíquota criopreservada, foram colhidos em animais jovens e adultos, com e sem raça definida e a ocorrência dos principais agentes etiológicos virais (BVDV, VSRB e BoHV) e bacterianos (Mannheimia haemolytica, Pasteurella multocida e micoplasma sp.) causadores de broncopneumonia em bovinos foram pesquisadas. Dos cultivos bacterianos específicos, as aliquotas criopreservada apresentaram 12/96 amostras positivas para mollicutes enquanto os cutivos utilizando as hastes foram 11/96 amostras positivas para P. multocida. No cultivo da P. multocida também foram encontradas Bacillus sp., Staphylococcus sp., Escherichia coli, Klebsiella oxytoca, Pseudomonas aeruginosa e Klebsiella pneumoniae sem que houvesse a ocorrência de M. haemolytica. PCR foi utilizada para detectar a classe mollicutes (68/96) e as espécies M. bovis (2/68), M. dispar (26/68) e M. mycoides subsp. mycoides (0/68). A técnica de vírus neutralização mostrou os seguintes resultados: 31/56 animais soreagentes para BVDV, 35/57 com BoVH e 54/58 com VSRB. Para análise estatística foi utilizado o teste do qui-quadrado em blocos ao acaso. Houve associação entre a presença de broncopneumonia em animais com BVDV (P=0,080) e com P. multocida (P=0,076). Animais da raça holandesa foram associados a presença de broncopneumonia (p= 0,095). Jovens com menos de 12 meses foram associados a presença de mollicutes (P=0,0177). A presença de som submaciço a percussão do gradil toráxico foi associado a ausência de mollicutes (P=0,025). A ausência de M. dispar foi associado a presença de sibilo (P=0,076) e de estertor úmido (P=0,046) na ausculta pulmonar. Houve uma chance maior de 5,5 vezes de apresentar tosse na presença de P. multocida enquanto na sua ausência hà uma maior chance dos animais apresentarem secreção mucosa (7,3) e mucopurulento (5,8), som submaciço (9,6), crepitação fina (13) e estertor úmido (5). Houve associação entre a ausência de M. bovis e a presença de som claro a percussão do gradil toráxico dos bovinos examinados. / Respiratory diseases are considered a serious problem of sanity, causing huge economic losses due to high rates of morbidity and mortality, accounting for 10-40% of deaths in adults and 17% of deaths during the perinatal period. The objective of this study was to determine the occurrence of agents and seek associations between agents and bronchopneumonia and the association of clinical signs with these agents. They were evaluated clinically and data collected in own record 96 animals of two herds of Bragança Paulista region and Mococa both in São Paulo. Whole blood and washed tracheobronchial were harvested in young and adult animals, Holstein and undefined breed and the occurrence of major viral etiologic agents (BVDV, BRSV and BoHV) and bacterial (Mannheimia haemolytica, Pasteurella multocida and Mycoplasma sp.) causing bronchopneumonia in cattle were surveyed. The specific bacterial cultures showed positive samples Mollicutes 12/96 and 11/96 samples positive for P. multocida. In the cultivation of P. multocida were also found Bacillus sp., Staphylococcus sp., Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Pseudomonas aeruginosa and there was no occurrence of M. haemolytica. PCR was used to detect the class Mollicutes (68/96) and the species M. bovis (2/68), M. dispar (26/68) and M. mycoides subsp. mycoides (0/68). The virus neutralization technique showed the following results: 31/56 animals BVDV positive, 35/57 and 54/58 with BoVH with BRSV. Statistical analysis was performed using the chi-square test in blocks. There is an association between bronchopneumonia in animals and the presence of BVDV (p = 0.080) and P. multocida (P = 0.076). Holstein animals were associated with the presence of bronchopneumonia (p = 0.095). Calves under 12 months were associated with the presence of Mollicutes (P = 0.0177). The sound submassive present during percussion of the thoracic cage was associated with the absence of Mollicutes (P = 0.025). The absence of M. dispar was associated with presence of wheezing (P = 0.076) and wet rattle (P = 0.046) in lung auscultation. There was a greater chance of 5.5 times to present cough in the presence of P. multocida as in its absence there is a greater chance of the animals show mucous secretion (7.3) and mucopurulent (5.8), submassive sound (9.6 ), thin crackling (13) and wet rattle (5). There was an association between the absence of M. bovis and the presence of clear sound of chest cage percussion of the examined cattle.
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Ocorrência de doenças respiratórias causadas por bactérias e vírus em ovinos / Occurrence of respiratory diseases caused by bacteria and viruses in sheepFranco, Mariane Ferreira 12 July 2018 (has links)
O Brasil é o 18ª maior produtor de carne ovina e, apesar de ser em grande parte informal, é uma cultura crescente no país. Dentre as enfermidades infecciosas que acomete a ovinocultura a broncopneumonia é uma das mais recorrentes, no entanto, não há muitos estudos sobre esta enfermidade em pequenos ruminantes no Brasil. Por isso, esta pesquisa teve como objetivo verificar a ocorrência de doenças respiratórias no estado de São Paulo e Rio de Janeiro causadas por bactérias e vírus. Para a realização desse projeto foi utilizado a técnica de lavado traqueobrônquico transtraqueal e coleta de sangue total para obtenção do soro em 99 ovinos. Essas amostras foram submetidas a teste de virusneutralização para identificação de anticorpos contra vírus da Parainfluenza Tipo 3 (PI-3), Herpesvírus Bovino Tipo 1 (BoHV- 1), Vírus Sincicial Respiratório Bovino (BRSV) e Vírus da Diarreia Viral Bovina (VDVB). Utilização do teste de imunodifusão em gel de agarose e Eradikt® para detectar a presença de anticorpos contra Lentivírus de Pequenos Ruminantes (LVPR). Isolamento e identificação bioquímica para M. haemolytica e Pasteurella multocida. Cultivo e isolamento, identificação bioquímica e PCR foram testes utilizados para identificação de micoplasmas (Mycoplasma bovis, M. agalactiae, M. mycoides subsp. capri). Das 99 amostras coletadas, 33 foram de ovinos doentes e 66 de ovinos sadios. Não houve identificação de M. haemolytica e P. multocida, nem presença de anticorpos contra BoHV-1 e VDVB. No entanto, observou-se a prevalência de 52,52%, 48,48% e 21,87% de PI-3, BRSV e LVPR respectivamente. Em relação as bactérias aeróbias, houve maior frequência de isolamento de Bacillus sp. e Gram-negativas não fermentadoras. Apesar de identificar bactérias da classe Mollicutes em colônias isoladas em 23,28% das amostras, houve apenas uma identificação com os oligonucleotídeos utilizados, o M. mycoides subsp. capri, primeiro isolamento em ovinos no estado de São Paulo e Rio de Janeiro. Houve diferença estatística significativa (p<0,05) entre animais com broncopneumonia e manifestações clínicas como taquipneia, hipertermia, secreção nasal, tosse, dispneia, crepitação fina e ronco e entre animais com broncopneumonia e não realizam de quarentena e não separam animais doentes. A broncopneumonia envolve vários fatores, incluindo o manejo, agente infeccioso e a imunidade do animal. Por isso, é necessário conhecer todos esses aspectos e associá-los para uma melhor prevenção e tratamento. / Brazil is the 18th largest producer of sheep meat and, despite being largely informal, is a growing crop in the country. Among the infectious diseases that affect sheep production, bronchopneumonia is one of the most recurrent, however, there are not many studies on this disease in small ruminants in Brazil. Therefore, this research aimed to verify the occurrence of respiratory diseases in the state of São Paulo and Rio de Janeiro caused by bacteria and viruses. For the accomplishment of this project was used tracheobronchial lavage technique transtracheal and collection of whole blood to obtain serum in 99 sheep. These samples were submitted to virus neutralization test to identify antibodies against Parainfluenza Type 3 (PI-3), Bovine Herpesvirus Type 1 (BoHV-1), Bovine Respiratory Syncytial Virus (BRSV) and Bovine Viral Diarrhea Virus (BVDV). Use of the Eradikit ® and agarose gel immunodiffusion test to detect the presence of antibodies against Small Ruminant Lentiviruses (LVPR). Isolation and biochemical identification for M. haemolytica and Pasteurella multocida. Cultivation and isolation, biochemical identification and PCR were used to identify mycoplasmas (Mycoplasma bovis, M. agalactiae, M. mycoides subsp. Capri). Of the 99 samples collected, 33 were from diseased sheep and 66 from healthy sheep. There was no identification of M. haemolytica and P. multocida, nor presence of antibodies against BoHV-1 and BVDV. However, the prevalence of 52.52%, 48.48% and 21.87% of PI-3, BRSV and LVPR, respectively, was observed. In relation to aerobic bacteria, there was a higher frequency of isolation of Bacillus sp. and Gram-negative non-fermenters. Despite identifying Mollicute class bacteria in isolated colonies in 23.28% of the samples, there was only one identification with the oligonucleotides used, M. mycoides subsp. capri, first isolation in sheep in the state of São Paulo and Rio de Janeiro. There was a significant statistical difference (p <0.05) between animals with bronchopneumonia and clinical manifestations such as tachypnea, hyperthermia, nasal secretion, cough, dyspnea, fine crackling and snoring, and between animals with bronchopneumonia and quarantine and separation of diseased animals. Bronchopneumonia involves several factors, including management, infectious agent and the immunity of the animal. Therefore, it is necessary to know all these aspects and to associate them for a better prevention and treatment.
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Avaliação clínica e estudo da ocorrência de broncopneumonia em bovinos / Clinical evaluation and study of the occurence of bronchopneumonia in cattleBernardo Augusto França Dias de Oliveira 27 July 2015 (has links)
As doenças respiratórias são consideradas um sério problema de sanidade, causando grandes perdas econômicas devido aos altos índices de morbidade e mortalidade, sendo responsáveis por 10 a 40% das mortes em animais adultos e por 17% das mortes durante o período perinatal. O objetivo deste trabalho foi determinar a ocorrência de agentes e buscar associações entre agentes e a broncopneumonia e a associação de sinais clínicos com os agentes presentes. Foram avaliados clinicamente e os dados coletados em ficha própria 96 animais de dois rebanhos da região de Bragança Paulista e Mococa ambas no Estado de São Paulo. Sangue total e lavado traqueobrônquico instilado em haste flexível e o restante da alíquota criopreservada, foram colhidos em animais jovens e adultos, com e sem raça definida e a ocorrência dos principais agentes etiológicos virais (BVDV, VSRB e BoHV) e bacterianos (Mannheimia haemolytica, Pasteurella multocida e micoplasma sp.) causadores de broncopneumonia em bovinos foram pesquisadas. Dos cultivos bacterianos específicos, as aliquotas criopreservada apresentaram 12/96 amostras positivas para mollicutes enquanto os cutivos utilizando as hastes foram 11/96 amostras positivas para P. multocida. No cultivo da P. multocida também foram encontradas Bacillus sp., Staphylococcus sp., Escherichia coli, Klebsiella oxytoca, Pseudomonas aeruginosa e Klebsiella pneumoniae sem que houvesse a ocorrência de M. haemolytica. PCR foi utilizada para detectar a classe mollicutes (68/96) e as espécies M. bovis (2/68), M. dispar (26/68) e M. mycoides subsp. mycoides (0/68). A técnica de vírus neutralização mostrou os seguintes resultados: 31/56 animais soreagentes para BVDV, 35/57 com BoVH e 54/58 com VSRB. Para análise estatística foi utilizado o teste do qui-quadrado em blocos ao acaso. Houve associação entre a presença de broncopneumonia em animais com BVDV (P=0,080) e com P. multocida (P=0,076). Animais da raça holandesa foram associados a presença de broncopneumonia (p= 0,095). Jovens com menos de 12 meses foram associados a presença de mollicutes (P=0,0177). A presença de som submaciço a percussão do gradil toráxico foi associado a ausência de mollicutes (P=0,025). A ausência de M. dispar foi associado a presença de sibilo (P=0,076) e de estertor úmido (P=0,046) na ausculta pulmonar. Houve uma chance maior de 5,5 vezes de apresentar tosse na presença de P. multocida enquanto na sua ausência hà uma maior chance dos animais apresentarem secreção mucosa (7,3) e mucopurulento (5,8), som submaciço (9,6), crepitação fina (13) e estertor úmido (5). Houve associação entre a ausência de M. bovis e a presença de som claro a percussão do gradil toráxico dos bovinos examinados. / Respiratory diseases are considered a serious problem of sanity, causing huge economic losses due to high rates of morbidity and mortality, accounting for 10-40% of deaths in adults and 17% of deaths during the perinatal period. The objective of this study was to determine the occurrence of agents and seek associations between agents and bronchopneumonia and the association of clinical signs with these agents. They were evaluated clinically and data collected in own record 96 animals of two herds of Bragança Paulista region and Mococa both in São Paulo. Whole blood and washed tracheobronchial were harvested in young and adult animals, Holstein and undefined breed and the occurrence of major viral etiologic agents (BVDV, BRSV and BoHV) and bacterial (Mannheimia haemolytica, Pasteurella multocida and Mycoplasma sp.) causing bronchopneumonia in cattle were surveyed. The specific bacterial cultures showed positive samples Mollicutes 12/96 and 11/96 samples positive for P. multocida. In the cultivation of P. multocida were also found Bacillus sp., Staphylococcus sp., Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Pseudomonas aeruginosa and there was no occurrence of M. haemolytica. PCR was used to detect the class Mollicutes (68/96) and the species M. bovis (2/68), M. dispar (26/68) and M. mycoides subsp. mycoides (0/68). The virus neutralization technique showed the following results: 31/56 animals BVDV positive, 35/57 and 54/58 with BoVH with BRSV. Statistical analysis was performed using the chi-square test in blocks. There is an association between bronchopneumonia in animals and the presence of BVDV (p = 0.080) and P. multocida (P = 0.076). Holstein animals were associated with the presence of bronchopneumonia (p = 0.095). Calves under 12 months were associated with the presence of Mollicutes (P = 0.0177). The sound submassive present during percussion of the thoracic cage was associated with the absence of Mollicutes (P = 0.025). The absence of M. dispar was associated with presence of wheezing (P = 0.076) and wet rattle (P = 0.046) in lung auscultation. There was a greater chance of 5.5 times to present cough in the presence of P. multocida as in its absence there is a greater chance of the animals show mucous secretion (7.3) and mucopurulent (5.8), submassive sound (9.6 ), thin crackling (13) and wet rattle (5). There was an association between the absence of M. bovis and the presence of clear sound of chest cage percussion of the examined cattle.
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Bronchopneumonies infectieuses des jeunes bovins : de la complexité du microbiome aux particularités évolutives et cliniques de virus respiratoires encore méconnus. / Infectious bovine bronchopneumonia : deciphering the complex microbiomes and evolutionary and clinical features of yet unfamiliar virusesSalem, Elias 24 October 2018 (has links)
L’étiologie des bronchopneumonies infectieuses (BPI) des jeunes bovins est multifactorielle, mettant en cause des agents infectieux comme des bactéries, virus ou parasites, et également des facteurs de risques liés à la conduite d’élevage et à l’environnement. Dans cette thèse nous nous sommes intéressés aux virus respiratoires bovins. Nous avons étudié le virome de l’appareil respiratoire superficiel et profond des jeunes veaux atteints de BPI par des approches de séquençage à haut débit pour mieux caractériser les co-infections virales et identifier de nouveaux virus. Par ailleurs nous nous sommes intéressés au contexte dans lequel les virus opèrent en analysant la structure, la diversité, et le dynamisme du bactériote respiratoire chez des veaux sains et atteints de BPI. Les résultats suggèrent que de nombreux virus agissent en interactions et montrent une prédominance du coronavirus bovin (BCoV) dans les cavités nasopharyngées et également dans les poumons des veaux atteints de BPI. L’étude phylogénétique des BCoV isolés indique une ségrégation entre souches européennes d’une part et américaines et asiatiques d’autre part qui semble résulter d’un phénomène de recombinaison dans les années 1960-70. Par ailleurs un astrovirus bovin a été clairement détecté pour la première fois principalement dans les poumons de veaux atteints de BPI. L’analyse du microbiote indique, elle, une disparité écologique entre cavités superficielles et profondes et des interactions possibles entre agents pathogènes connus et différentes communautés bactériennes de la flore résidente. Enfin une partie des travaux a concerné le virus influenza D (IDV), un nouveau virus respiratoire bovin émergent récemment identifié en France. Lors d’une infection expérimentale chez des veaux nous avons démontré que IDV possède un pouvoir pathogène respiratoire modéré et qu’il module la réponse immunitaire innée du veau. Nous avons aussi confirmé le caractère ubiquiste d’IDV en démontrant sa circulation sur le continent africain. En conclusion, grâce à des méthodes de séquençages à haut débit ce travail a permis une meilleure description et caractérisation des virus respiratoires bovins et de leur environnement immédiat. Il ouvre des perspectives pour mieux comprendre le rôle des interactions virales dans la genèse des signes cliniques respiratoires. / Bovine infectious bronchopneumonia (BIP) is a complex syndrome that affects young bovines, with a multifactorial etiology often involving one or several viruses and bacteria favored by altered host immunity and disturbed environmental factors. First, using viral metagenomics sequencing tools, we explored the upper (URT) and lower (LRT) respiratory tract viromes of calves with BPI and identified unexpected viruses. In addition, in the same calves, we characterized the structure, diversities and dynamism of the bacterial communities. Results showed different patterns of interactions between the different components of the microbiomes. Among the many detected viruses, the bovine coronavirus (BCoV), showed the highest prevalence in both nasal and pulmonary cavities. Evolutionary and phylogenic analysis of the isolated BCoV strains indicated a clear segregation between European and American/Asian strains, which seems to have resulted from a recombination event during the 1960-70’s. Furthermore, a bovine astrovirus was detected for the first time in the lungs of BIP affected calves. A disparity was noticed in the bacterial community structures between the upper and lower respiratory cavities. Also, there were associations between the presences of certain bacterial taxa and known respiratory pathogens. Finally, a part of the work focused on the emerging influenza D virus (IDV) recently identified in France. Carrying an experimental infection, we showed that IDV has a moderate respiratory pathogenicity and can modulate the innate immune response of the calf. We also showed that IDV circulates in Africa thus confirming its global distribution. In conclusion, thanks to high throughput sequencing methods, this piece of work allowed for a detailed characterization of the bovine respiratory virome and its interacting environment, and further opened new perspectives for a better understanding of viral interactions in bovine BIP.
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Vergleichende Untersuchungen von BALB/c- und C57BL/6-Mäusen nach experimenteller Infektion mit Streptobacillus moniliformis oder Rodentibacter pneumotropicusFornefett, Juliane 21 May 2019 (has links)
Zielstellung: Ziel dieser kumulativen Dissertationsarbeit war die vergleichende Untersuchung der Wirtsantwort in verschiedenen Mauslinien nach Infektion mit Streptobacillus (S.) moniliformis und Rodentibacter (R.) pneumotropicus mit Anzeigeparametern für die Klinik, die Pathologie und die Immunantwort. Es sollten neue erregerspezifische enzyme-linked immunosorbent assays (ELISA) evaluiert und durch die Bestimmung der Immunglobulin G (IgG)-Subklassen mausstammspezifische Unterschiede in der Immunantwort aufgezeigt werden. Darüber hinaus waren Sentinelsysteme zu bewerten.
Material und Methoden: Es wurden BALB/c und C57BL/6-Mäuse intranasal mit S. moniliformis- oder R. pneumotropicus infiziert und mit Kontaktsentinels vergesellschaftet. Zusätzlich wurde benutzte Einstreu der Infektionskäfige auf Käfige mit nichtinfizierten CD1-Mäusen (Einstreu-Sentinels) übertragen. Die Infektionsgruppen wurden über 4 Wochen, die Sentinels mindestens 7 Wochen alle 12 Stunden klinisch untersucht und die Verläufe dokumentiert. Am Ende der Experimente bzw. bei Erreichen spezifischer Abbruchkriterien wurden die Mäuse tierschutzgerecht euthanasiert und definierte Organproben für pathohistologische und bakteriologische Untersuchungen gewonnen. Die Erreger wurden dabei massenspektrometrisch sowie mittels polymerase chain reaction (PCR) differenziert und in Proben des Respirationstraktes quantitativ erfasst. Erregerspezifische Antikörper wurden in tracheonasaler Spülflüssigkeit und im Serum in eigens etablierten ELISA‘s auf Basis von Ganzzellextrakten bestimmt. Weiterhin erfolgte die Messung des Verhältnisses der IgG-Subtypen IgG1 und IgG2 im ELISA.
Ergebnisse: Der Infektionsversuch mit S. moniliformis bestätigte mit einer Mortalität von 75% die bekannte hohe Infektionsanfälligkeit der C57BL/6-Mäuse im Gegensatz zu BALB/c, die keine Krankheitsanzeichen entwickelten. Die wichtigsten pathologischen Manifestationen waren eitrig-nekrotisierende Lymphadenitiden und Pneumonien in Verbindung mit der Reisolation des Infektionsstammes. Mithilfe des etablierten ELISA‘s gelang der Nachweis erregerspezifischer IgG-Antikörper im Serum der Tiere beider Linien. Bei den Kontaktsentinels konnte, bis auf eine Ausnahme, weder kulturell, noch serologisch eine Infektion nachgewiesen werden. Gleiches gilt für alle Einstreu-Sentinels. Molekularbiologisch wurde aber Erreger-DNA in den Lungen der Sentinels festgestellt. Die Infektion mit einem R. pneumotropicus Stamm, welcher genotypisch positiv für alle drei bekannten RTX-Toxine dieses Erregers war, führte zu einer unerwartet hohen Morbidität und Mortalität in beiden Mauslinien. In frühzeitig euthanasierten Tieren beider Linien konnten katarrhalisch-eitrige bis nekrotisierende Bronchopneumonien sowie eine Dissemination des Belastungsstammes in zahlreiche innere Organe nachgewiesen werden. In überlebenden Tieren beider Linien wurde eine deutliche Kolonisation respiratorischer Schleimhäute mit dem Belastungsstamm trotz z.T. hoher mukosaler IgA-Spiegel und Serokonversion im Blut nachgewiesen. Überlebende C57BL/6 Mäuse zeigten eine signifikant niedrigere Bakterienlast in inneren Organen als BALB/c Mäuse. In allen Kontaktsentinels, aber nicht in einem einzigen Einstreu-Sentinel, konnte kulturell und indirekt serologisch eine Infektion mit R. pneumotropicus nachgewiesen werden. Die Bestimmung der IgG-Subklassen in den Seren der C57BL/6-Mäuse beider Infektionsstudien ergab eine Verschiebung des Verhältnisses IgG2/IgG1 von unter 0,8 vor zu über 1,6 nach Infektion. Dies weist auf eine T-Helferzell (Th) 1-dominierte Immunantwort hin. BALB/c-Mäuse behielten dagegen ein Verhältnis unter 0,8 auch nach der Infektion bei, sodass auf eine Th2- Antwort zu schließen war.
Schlussfolgerungen: Sowohl für S. moniliformis als auch für R. pneumotropicus konnten Tiermodelle mit diversen Anzeigeparametern etabliert werden, welche für Folgestudien zur Pathogenese oder Immunprophylaxe genutzt werden können. Für beide Erreger wurden neue sensitive und spezifische ELISA-Protokolle in die Diagnostik eingeführt. Kontaktsentinels, aber nicht Einstreu-Sentinels, sind gut geeignet, um R. pneumotropicus-Infektionen nachzuweisen. Die beobachtete stammspezifische Klinik, Pathologie und Immunantwort der C57BL/6-Mäuse nach experimenteller S. moniliformis-Infektion sprach für eine pathologische Th1-Immunantwort. Im Gegensatz dazu war im R. pneumotropicus – Infektionsversuch die Th1-Immunantwort der C57BL/6-Mäuse mit einer effektiveren Reduktion des Erregers in inneren Organen assoziiert. Die unerwartet hohe Morbidität und Mortalität im R. pneumotropicus –Infektionsversuch weist auf eine besonders hohe Virulenz des eingesetzten Stammes hin, sodass in dieser Arbeit erstmalig ein septikämischer Verlauf in Wildtyp-Mäusen nach intranasaler R. pneumotropicus-Infektion nachgewiesen werden konnte.:Inhaltsverzeichnis (I)
Abkürzungsverzeichnis (III)
1 Einleitung (1)
2 Literatur (3)
2.1 Streptobacillus moniliformis (3)
2.1.1 Allgemeine Charakteristika (3)
2.1.2 Differenzierung von Streptobacillus spp.(4)
2.1.3 Serologische Methoden zum indirekten Nachweis einer Streptobacillus
moniliformis - Infektion (5)
2.1.4 Epidemiologie der durch Streptobacillus moniliformis hervorgerufenen
Zoonose (6)
2.1.5 Klinik und Pathologie der Streptobacillus moniliformis-Infektion bei
Nagetieren (8)
2.1.6 Klinik und Pathologie der Streptobacillus moniliformis-Infektion in Menschen
und anderen Nebenwirten (9)
2.1.7 Pathogenese und Virulenzfaktoren (10)
2.1.8 Prävalenz in Nagern (11)
2.1.9 Sanierung Streptobacillus moniliformis infizierter Nagetierbestände und
Prävention (12)
2.2 Rodentibacter (R.) pneumotropicus und heylii (Pasteurella (P.) pneumotropica
Biotyp Jawetz und Heyl) (14)
2.2.1 Allgemeine Charakteristika (14)
2.2.2 Ursprüngliche Einteilung in Biotypen und Reklassifikation zu Rodentibacter
spp. (14)
2.2.3 Differenzierung von Rodentibacter spp. (15)
2.2.4 Serologische Methoden zum indirekten Nachweis einer Rodentibacter-
Infektion (15)
2.2.5 Übertragung (15)
2.2.6 Klinik und Pathologien der Rodentibacter-Infektion in Nagern (16)
2.2.7 Pathogenese und Virulenzfaktoren (18)
2.2.8 Prävalenz (19)
2.2.9 Sanierung Rodentibacter pneumotropicus infizierter Nagetierbestände
und Prävention (20)
2.3 Mäuse als Versuchstiere (22)
2.3.1 Inzucht-Stämme (22)
2.3.1.1 Merkmale, Verwendung und Historie der BALB/c-Wildtypmäuse (22)
2.3.1.2 Merkmale, Verwendung und Historie der C57BL/6-Wildtypmäuse (23)
2.3.1.3 Unterschiede der Immunreaktionen in C57BL/6- und der BALB/c-
Mäusen (23)
2.3.2 Auszucht-Stämme (24)
2.3.2.1 Merkmale, Verwendung und Historie der CD1-Wildtypmäuse (24)
2.4 Gesundheitsmonitoring in Labortierhaltungen (25)
2.4.1 Empfehlungen der Federation of Laboratory Animal Science Associations
(FELASA) (25)
2.4.2 Sentinelsysteme für das Gesundheitsmonitoring in
Labormausbeständen (26)
3 Publikationen (28)
3.1 Fornefett J, Krause J, Klose K, Fingas F, Hassert R, Eisenberg T, Grunwald T,
Müller U, Schrödl W, Baums CG. Comparative analysis of clinics, pathologies
and immune responses in BALB/c and C57BL/6J mice infected with
Streptobacillus moniliformis. Microbes and Infection. 2018;20(2):101-110 (28)
3.2 Fornefett J, Krause J, Klose K, Fingas F, Hassert R, Benga L, Grunwald T,
Müller U, Schrödl W, Baums CG. Comparative analysis of humoral immune
responses and pathologies of BALB/c and C57BL/6 wildtype mice
experimentally infected with a highly virulent Rodentibacter pneumotropicus
(Pasteurella pneumotropica) strain. BMC Microbiology. 2018;18(1):45 (39)
4 Übergreifende Diskussion (51)
5 Zusammenfassung (59)
6 Summary (61)
7 Literaturverzeichnis (63)
7.1 Fachliteratur (63)
7.2 Internet (76)
7.3 Gesetzestexte (78)
Anhang (79)
i Ergänzende Abbildungen (79)
ii Ergänzendes Material zu 3.1 “Comparative analysis of clinics, pathologies and
immune responses in BALB/c and C57BL/6J mice infected with Streptobacillus
moniliformis” (81)
iii Ergänzendes Material zu 3.2 “Comparative analysis of humoral immune
responses and pathologies of BALB/c and C57BL/6 wildtype mice
experimentally infected with a highly virulent Rodentibacter pneumotropicus
(Pasteurella pneumotropica) strain” (83)
Liste mit weiteren Veröffentlichungen
Danksagung / Objective: Aim of this cumulative doctoral thesis was the comparative analysis of the host response in different mice strains infected with Streptobacillus (S.) moniliformis and Rodentibacter (R.) pneumotropicus with readout parameters for clinics, pathology and immune response. New pathogen specific enzyme linked immunosorbent assay’s (ELISA) were evaluated. Differentiation of immunoglobulin (Ig) G subclasses was conducted to reveal differences in the immune response between the two mice strains. Furthermore, sentinel systems were assessed.
Materials and methods: BALB/c and C57BL/6 mice were infected intranasally with S. moniliformis or R. pneumotropicus and housed together with contact sentinels. Soiled bedding from infection cages was transmitted to cages with uninfected CD1 mice (bedding sentinels). Infection groups were observed for 4 weeks, sentinels for at least 7 weeks and the clinical course was documented. At the end of the experiments or when predefined termination criteria were reached, animals were humanely killed. Predefined organ samples were collected for pathohistological and bacteriological screenings. Pathogens were differentiated via mass spectrometry and via polymerase chain reaction (PCR). The specific bacterial load was quantified in samples of the respiratory tract. Pathogen-specific antibodies were detected in tracheonasal lavages and sera using newly established ELISAs based on whole cell extracts. Determination of the ratios of the IgG subtypes (IgG1 to IgG2) was conducted using ELISAs.
Results: The S. moniliformis experiment confirmed the known high susceptibility of C57BL/6 mice with a mortality of 75%. This was in contrast to BALB/c, which developed no signs of illness. The major pathologies were purulent-necrotizing inflammations of the lymph nodes and the lung associated with detection of the challenge strain. Specific IgG-antibodies were detected in sera of both mice strains by the newly established ELISAs. In contact and bedding sentinels the infection was not detected by culture or indirectly by serology, except for one contact sentinel. However, pathogen DNA was detectable in the lungs of these animals via PCR. The infection with the R. pneumotropicus strain, which is genotypically positive for all 3 known RTX toxins of this pathogen, leaded to an unexpected high morbidity and mortality in both mice strains. In early losses a catharal-purulent to necrotizing bronchopneumonia as well as dissemination of the challenge strain in various inner organs was recorded. Efficient colonization of the respiratory mucosa through the challenge strain was detected in survivors of both lines despite high mucosal IgA levels and seroconversion in the blood. Surviving C57BL/6 mice showed a significant lower bacterial burden in inner organs than BALB/c. All contact sentinels were culturally and serologically positive for R. pneumotropicus infection in contrast to all bedding sentinels.
Differentiation of IgG subclasses in sera of C57BL/6 mice of both experiments revealed a shift of the IgG2/IgG1 ratio from 0.8 prior to infection to 1.6 post infection suggesting a T helper (Th) 1-prone immune response. BALB/c mice remained under 0.8 after infection indicating a Th2-prone immune response.
Conclusions: New animal models with various readout parameters were established for both S. moniliformis and R. pneumotropicus. These models can be used in future studies on pathogenesis and immunoprophylaxis. Sensitive und specific ELISA-protocols were established for both pathogens. Contact sentinels but not bedding sentinels are appropriate for detection of R. pneumotropicus-infections. The observed distinct clinic, pathology and immune response of C57BL/6 mice experimentally infected with S. moniliformis indicated on the one hand a pathological Th1 immune response. On the other hand, the Th1 response of C57BL/6 mice to R. pneumotropicus infection was associated with a more efficient clearance of the challenge strain in internal organs. The unprecedented high morbidity and mortality in the R. pneumotropicus experiment indicates high virulence of this strain. Accordingly, this work revealed for the first time septicaemia in wildtype mice after intranasal R. pneumotropicus-infection.:Inhaltsverzeichnis (I)
Abkürzungsverzeichnis (III)
1 Einleitung (1)
2 Literatur (3)
2.1 Streptobacillus moniliformis (3)
2.1.1 Allgemeine Charakteristika (3)
2.1.2 Differenzierung von Streptobacillus spp.(4)
2.1.3 Serologische Methoden zum indirekten Nachweis einer Streptobacillus
moniliformis - Infektion (5)
2.1.4 Epidemiologie der durch Streptobacillus moniliformis hervorgerufenen
Zoonose (6)
2.1.5 Klinik und Pathologie der Streptobacillus moniliformis-Infektion bei
Nagetieren (8)
2.1.6 Klinik und Pathologie der Streptobacillus moniliformis-Infektion in Menschen
und anderen Nebenwirten (9)
2.1.7 Pathogenese und Virulenzfaktoren (10)
2.1.8 Prävalenz in Nagern (11)
2.1.9 Sanierung Streptobacillus moniliformis infizierter Nagetierbestände und
Prävention (12)
2.2 Rodentibacter (R.) pneumotropicus und heylii (Pasteurella (P.) pneumotropica
Biotyp Jawetz und Heyl) (14)
2.2.1 Allgemeine Charakteristika (14)
2.2.2 Ursprüngliche Einteilung in Biotypen und Reklassifikation zu Rodentibacter
spp. (14)
2.2.3 Differenzierung von Rodentibacter spp. (15)
2.2.4 Serologische Methoden zum indirekten Nachweis einer Rodentibacter-
Infektion (15)
2.2.5 Übertragung (15)
2.2.6 Klinik und Pathologien der Rodentibacter-Infektion in Nagern (16)
2.2.7 Pathogenese und Virulenzfaktoren (18)
2.2.8 Prävalenz (19)
2.2.9 Sanierung Rodentibacter pneumotropicus infizierter Nagetierbestände
und Prävention (20)
2.3 Mäuse als Versuchstiere (22)
2.3.1 Inzucht-Stämme (22)
2.3.1.1 Merkmale, Verwendung und Historie der BALB/c-Wildtypmäuse (22)
2.3.1.2 Merkmale, Verwendung und Historie der C57BL/6-Wildtypmäuse (23)
2.3.1.3 Unterschiede der Immunreaktionen in C57BL/6- und der BALB/c-
Mäusen (23)
2.3.2 Auszucht-Stämme (24)
2.3.2.1 Merkmale, Verwendung und Historie der CD1-Wildtypmäuse (24)
2.4 Gesundheitsmonitoring in Labortierhaltungen (25)
2.4.1 Empfehlungen der Federation of Laboratory Animal Science Associations
(FELASA) (25)
2.4.2 Sentinelsysteme für das Gesundheitsmonitoring in
Labormausbeständen (26)
3 Publikationen (28)
3.1 Fornefett J, Krause J, Klose K, Fingas F, Hassert R, Eisenberg T, Grunwald T,
Müller U, Schrödl W, Baums CG. Comparative analysis of clinics, pathologies
and immune responses in BALB/c and C57BL/6J mice infected with
Streptobacillus moniliformis. Microbes and Infection. 2018;20(2):101-110 (28)
3.2 Fornefett J, Krause J, Klose K, Fingas F, Hassert R, Benga L, Grunwald T,
Müller U, Schrödl W, Baums CG. Comparative analysis of humoral immune
responses and pathologies of BALB/c and C57BL/6 wildtype mice
experimentally infected with a highly virulent Rodentibacter pneumotropicus
(Pasteurella pneumotropica) strain. BMC Microbiology. 2018;18(1):45 (39)
4 Übergreifende Diskussion (51)
5 Zusammenfassung (59)
6 Summary (61)
7 Literaturverzeichnis (63)
7.1 Fachliteratur (63)
7.2 Internet (76)
7.3 Gesetzestexte (78)
Anhang (79)
i Ergänzende Abbildungen (79)
ii Ergänzendes Material zu 3.1 “Comparative analysis of clinics, pathologies and
immune responses in BALB/c and C57BL/6J mice infected with Streptobacillus
moniliformis” (81)
iii Ergänzendes Material zu 3.2 “Comparative analysis of humoral immune
responses and pathologies of BALB/c and C57BL/6 wildtype mice
experimentally infected with a highly virulent Rodentibacter pneumotropicus
(Pasteurella pneumotropica) strain” (83)
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Danksagung
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