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Doctoral Thesis
DOI
https://doi.org/10.11606/T.10.2021.tde-28052021-134553
Document
Author
Full name
Maria Luana Alves
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2021
Supervisor
Committee
Oliveira, Trícia Maria Ferreira de Sousa (President)
Bresciani, Katia Denise Saraiva
Dellova, Deise Carla Almeida Leite
Nakaghi, Andrea Cristina Higa
Soares, Rodrigo Martins
Title in Portuguese
Identificação e caracterização de tripanossomatídeos que infectam gatos domésticos (Felis catus) de área endêmica para leishmaniose visceral
Keywords in Portuguese
Crithidia fasciculata
L. infantum
Lu. Longipalpis
Gatos
Abstract in Portuguese
Um total de 100 gatos procedentes de dois abrigos de animais domésticos de Ilha Solteira, SP, participaram do estudo. Pelas técnicas sorológicas 74% e 34% dos gatos apresentaram anticorpos anti- Leishmania spp. pela Reação de Imunofluorescência Indireta (RIFI) e o Ensaio Imunoenzimático Indireto (ELISA), respectivamente. No parasitológico direto, dois animais (2%) apresentaram formas amastigotas no interior de macrófagos de linfonodo e medula óssea, enquanto na hemocultura, oito gatos (8%) apresentaram flagelados. Pela Reação em Cadeia da Polimerase (PCR) das amostras de sangue dos gatos do estudo, oito (8%) foram positivos por esta técnica e verificou-se a presença de sequências de DNA com 100% de identidade com L. infantum em cinco gatos, e sequências de DNA com ao menos 80% de identidade com L. major em dois animais. Dois dos quatro animais positivos pela cultura de aspirado de linfonodo e medula óssea apresentaram fragmentos de DNA de tripanossomatídeos, e quando sequenciados, revelaram a presença de DNA de L. infantum e Crithidia fasciculata. Um mesmo gato apresentou três espécies de tripanossomatídeos (L. infantum, C. fasciculata e L. major), detectadas pelo sequenciamento de amostras de DNA de sangue e DNA de cultura de aspirado de linfonodo, e caracterização de espécie pela Eletroforese Enzimática Multilocus (MLEE) de cepa obtida do isolado de aspirado de linfonodo, respectivamente. A maioria dos animais do estudo (74%) apresentou ao menos uma alteração clínica. Em relação aos parâmetros hematológicos, o grupo dos gatos positivos para L. infantum apresentou redução das plaquetas (p=0,01076 p<0,05), quando comparado ao grupo dos animais negativos, mostrando uma associação entre infecção por L. infantum e trombocitopenia. Sessenta animais (60%) foram positivos pela reação intradérmica de Montenegro (RIM), mas a maioria (80%) dos gatos positivos para L. infantum pelos testes moleculares e parasitológicos, não respondeu à RIM. Das sete espécies de flebotomíneos capturadas no estudo, Lu. longipalpis foi a mais frequente, sendo encontrado DNA de L. infantum em uma das fêmeas.
Title in English
Identification and characterization of the trypanosomatids infecting domestic cats (Felis catus) in endemic area for visceral leishmaniasis
Keywords in English
Crithidia fasciculata
L. infantum
Lu. longipalpis
Cats
Abstract in English
A total of 100 cats from two domestic animal shelters in Ilha Solteira, SP, participated in the study. By serological techniques 74% and 34% of cats presented antibodies against Leishmania spp. by Immunofluorescence Antibody Test (IFAT) and Indirect Enzyme-Linked Immunosorbent Assay (ELISA), respectively. In the parasitological test, two animals (2%) had amastigote forms inside lymph node and bone marrow macrophages, while in blood culture, eight cats (8%) had flagellates. By the Polymerase Chain Reaction (PCR) of the blood samples of the cats in the study, eight (8%) were positive by this technique and the presence of DNA sequences with 100% identity with L. infantum was verified in five cats, and DNA sequences with at least 80% identity with L. major in two animals. Two of the four animals positive for the lymph node and bone marrow aspirate culture presented DNA fragments of trypanosomatids, and when sequenced, they revealed the presence of L. infantum and Crithidia fasciculata DNA. The same cat presented three species of trypanosomatids (L. infantum, C. fasciculata and L. major), detected by sequencing blood DNA samples and culture DNA from lymph node aspirate, and species characterization by Multilocus Enzymatic Electrophoresis (MLEE) strain obtained from the lymph node aspirate isolate, respectively. Most of the animals in the study (74%) had at least one clinical alteration. Regarding hematological parameters, the group of cats positive for L. infantum showed a reduction in platelets (p = 0.01076 p <0.05), when compared to the group of negative animals, showing an association between infection by L. infantum and thrombocytopenia. Sixty (60%) animals were Montenegro Skin Test (MST) positive, but the majority (80%) of cats positive for L. infantum by molecular and parasitological tests, did not respond to MST. Of the seven species of sandflies captured in the study, Lu. longipalpis was the most frequent, and DNA of L. infantum was found in one of the females.
 
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Publishing Date
2021-07-23
 
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