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Master's Dissertation
DOI
10.11606/D.10.2018.tde-01082018-111738
Document
Author
Full name
Julieta Canjimba Porto Lucas Alexandre
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2018
Supervisor
Committee
Cogliati, Bruno (President)
Sanches, Daniel Soares
Timenetsky, Jorge
Title in Portuguese
Avaliação anatomopatológica, imunohistoquímica e molecular da Pleuropneumonia Contagiosa Bovina em animais sacrificados em matadouros no Huambo/Angola 
Keywords in Portuguese
Anatomopatológico
Bovino
Lesões
Pleuropneumonia Contagiosa Bovina
Abstract in Portuguese
A Pleuropneumonia Contagiosa Bovina (PPCB) é uma enfermidade respiratória grave, causada pelo Mycoplasma mycoides subsp mycoides Small Colony (MmmSC), sendo de primeira ordem no quadro nosológico em Angola, com alta prevalência e grandes repercussões econômicas na pecuária angolana. Objetivando caracterizar os achados anatomopatológicos e a resposta inflamatória na ocorrência natural da PPCB, realizou-se a coleta de amostras de pulmão e linfonodos regionais em 50 bovinos com lesões macroscópicas compatíveis com a doença e abatidos na cidade de Huambo, Angola. As amostras teciduais foram fixadas em formol 10% e incluídas em parafina para avaliação histopatológica, pela técnica de hematoxilina-eosina, e detecção do agente infeccioso por PCR a partir do DNA total extraído do tecido pulmonar parafinado. A caracterização da resposta inflamatória foi avaliada por imunohistoquímica, utilizando-se o método de detecção por polímero para os marcadores de linfócitos T (CD3) e B (Pax5), assim como as interleucinas (IL)-1β e 4. Após avaliação molecular, 7 amostras foram excluídas por não apresentarem DNA viável e 16/43 (37,2%) animais foram positivos para o MmmSC pelo PCR. Dos animais positivos, a lesão macroscópica mais frequente foi a aderência (7/16, 43,8%); enquanto a lesão microscópica mais frequente foi a hiperplasia linfoide (9/16, 56,2%). Houve associação significativa entre a positividade dos animais e a presença da alteração nas lesões de aderência (p=0,047), pulmão marmoreado (p=0,012), sequestro (p=0,001) e fibrose pulmonares (p<0,05). O pulmão esquerdo e lobo diafragmático esquerdo foram frequentemente afetados na doença, porém sem diferença significativa. Os resultados imunohistoquímicos demonstraram marcação focal para linfócitos B e T, assim como imunoexpressão de IL-1β e IL-4 em intensidade fraca a moderada na maioria dos casos analisados. Os resultados demostraram que a PPCB está presente em bovinos na região estudada, porém com lesões anatomopatológicas e resposta inflamatória inespecíficas. Estes resultados ressaltam a importância da avaliação integrada de dados epidemiológicos, anatomopatológicos, imunológicos e moleculares para estabelecer um diagnóstico preciso e definitivo da PPCB nestes animais.
Title in English
Anatomopathological, immunohistochemical and molecular evaluation of Bovine Contagious Pleuropneumonia in animals slaughtered in slaughterhouses in Huambo/Angola
Keywords in English
Anatomopathological
Bovine
Bovine Contagious Pleuropneumonia
Lesions
Abstract in English
Bovine Contagious Pleuropneumonia (PPCB) is a serious respiratory disease caused by Mycoplasma mycoides subsp mycoides Small Colony (MmmSC) , being of first order in the nosological framework in Angola, with high prevalence and great economic repercussions in Angolan livestock. Aiming to characterize the anatomopathological findings and the inflammatory response in the natural occurrence of PPCB, samples of lung and regional lymph nodes were collected in 50 cattle with macroscopic lesions compatible with the disease and slaughtered in the city of Huambo, Angola. Tissue samples were fixed in 10% formalin and embedded in paraffin for histopathological evaluation by the hematoxylin-eosin technique and detection of the infectious agent by PCR from the total DNA extracted from the paraffin-shaped lung tissue. The characterization of the inflammatory response was evaluated by immunohistochemistry, using the polymer detection method for T (CD3) and B (Pax5) lymphocytes, as well as interleukins (IL) -1β and 4. After molecular evaluation, 7 samples were excluded because they did not present viable DNA and 16/43 (37.2%) animals were positive for MmmSC by PCR. Of the positive animals, the most frequent macroscopic lesion was adhesion (7/16, 43.8%); while the most frequent microscopic lesion was lymphoid hyperplasia (9/16, 56.2%). There was a significant association between the positivity of the animals and the presence of changes in adhesion lesions (p = 0.047), marbled lung (p = 0.012), pulmonary sequestra (p = 0.001) and fibrosis (p <0.05). The left lung and left diaphragmatic lobe were frequently affected in the disease, but without significant difference. Immunohistochemical results demonstrated focal labeling for B and T lymphocytes as well as weak to moderate intensity of IL-1β and IL-4 immunoexpression in most of the analyzed cases. The results showed that CPBP is present in cattle in the studied region, but with nonspecific anatomopathological lesions and inflammatory response. These results highlight the importance of the integrated evaluation of epidemiological, anatomopathological, immunological and molecular data to establish an accurate and definitive diagnosis of CBPP in these animals.
 
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Publishing Date
2018-09-14
 
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