SEBASTIÁN ALEJANDRO MUÑOZ LEAL Study on Anaplasma, Borrelia, Coxiella, Ehrlichia ...

200 Pages·2017·26.13 MB·English

SEBASTIÁN ALEJANDRO MUÑOZ LEAL

Study on Anaplasma, Borrelia, Coxiella, Ehrlichia, Rickettsia and
Hepatozoon agents in ticks (Acari: Ixodoidea: Argasidae, Ixodidae)
from Chile, and a taxonomic study on Ornithodoros capensis sensu
lato (Acari: Argasidae) in South America

São Paulo
2017

SEBASTIÁN ALEJANDRO MUÑOZ LEAL

Study on Anaplasma, Borrelia, Coxiella, Ehrlichia, Rickettsia and
Hepatozoon agents in ticks (Acari: Ixodoidea: Argasidae, Ixodidae)
from Chile, and a taxonomic study on Ornithodoros capensis sensu
lato (Acari: Argasidae) in South America

Thesis submitted to the Postgraduate
Program in Experimental Epidemiology
Applied to Zoonoses of the School of
Veterinary Medicine and Animal Science of
the University of São Paulo to obtain the
Doctor’s degree in Sciences.

Departamento:
Preventive Veterinary Medicine and Animal
Health
Área de concentração:
Experimental Epidemiology Applied to
Zoonoses
Orientador:
Prof. Dr. Marcelo Bahia Labruna

De acordo:______________________
Orientador

São Paulo
2017
Obs: A versão original se encontra disponível na Biblioteca da FMVZ/USP
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DADOS INTERNACIONAIS DE CATALOGAÇÃO NA PUBLICAÇÃO

(Biblioteca Virginie Buff D’Ápice da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo)

T. 3580 Muñoz Leal, Sebastián Alejandro
FMVZ Study on Anaplasma, Borrelia, Coxiella, Ehrlichia, Rickettsia and Hepatozoon agents

in ticks (Acari: Ixodoidea: Argasidae, Ixodidae) from Chile, and a taxonomic study on
Ornithodoros capensis sensu lato (Acari: Argasidae) in South America / Sebastián
Alejandro Muñoz Leal. -- 2017.

200 p. : il.

Título traduzido: Pesquisa dos agentes Anaplasma, Borrelia, Coxiella, Ehrlichia,

Rickettsia e Hepatozoon em carrapatos (Acari: Ixodoidea: Argasidae, Ixodidae) do Chile e
estudo taxonômico de Ornithodoros capensis sensu lato (Argasidae) na América do Sul.

Tese (Doutorado) - Universidade de São Paulo. Faculdade de Medicina Veterinária e
Zootecnia. Departamento de Medicina Veterinária Preventiva e Saúde Animal, São Paulo,
2017.

Programa de Pós-Graduação: Epidemiologia Experimental Aplicada às Zoonoses.
Área de concentração: Epidemiologia Experimental Aplicada às Zoonoses..
Orientador: Prof. Dr. Marcelo B. Labruna.

1. Chile. 2. Ticks. 3. Tick-borne microorganisms. 4. PCR. 5.Phylogenetic analysis. I.
Título.

EVALUATION FORM

Author: MUÑOZ LEAL, Sebastián Alejandro

Title: Study on Anaplasma, Borrelia, Coxiella, Ehrlichia, Rickettsia and
Hepatozoon agents in ticks (Acari: Ixodoidea: Argasidae, Ixodidae) from
Chile, and a taxonomic study on Ornithodoros capensis sensu lato
(Acari: Argasidae) in South America.

Thesis submitted to the Postgraduate
Program in Experimental Epidemiology
Applied to Zoonoses of the School of
Veterinary Medicine and Animal Science
of the University of São Paulo to obtain
the Doctor’s degree in Sciences

Date: _____/_____/_____

Committee Members

Prof. _______________________________________________________________
Institution:__________________________ Decision: _________________________

Prof. _______________________________________________________________
Institution: __________________________ Decision: _________________________

Prof. _______________________________________________________________
Institution: __________________________ Decision: _________________________

Prof. _______________________________________________________________
Institution:__________________________ Decision: _________________________

Prof. _______________________________________________________________
Institution:__________________________ Decision: _________________________
DEDICATION

This work is dedicated to my parents, Ximena and René,
who early introduced me to nature
and encouraged me to keep studying it.
ACKNOWLEDGEMENTS

The current study represents the combined efforts of an international group of
tick researchers and I am most grateful for the time that they have invested in order
to contribute to this work.
In first place, I would like to thank Marcelo B. Labruna for the numerous
advices provided until the last analyses in the course of this study. Without his
expertise, this work would be likely to include many inaccurate statements. Long and
fruitful talks on soft tick systematics with him guided a great part of this work.
Extensive discussions on Argasidae were also shared with José M. Venzal, who in
two occasions joined the fieldwork group, and actively participated in the expedition
to Northern Chile and Southern Peru (2016), and to Northern Peru (2017).
The collection of the material and obtention of results in the initial phases of
this work would never have been achieved without the strategic assistance of Daniel
González-Acuña, who also enthusiastically contributed in fieldwork, and in the
organization of the expeditions for the collection of material in several Islands in Chile
(2015), and to the Southern region of Peru (2016). Much appreciated assistance was
received from Thiago F. Martins, Nicolás Ferrada, Cecilia Aguayo, Iván Torres,
Isabella Fontana, and Igor C. L. Acosta during fieldwork in Chile, and from Hermes
Ribeiro Luz, Débora Panizzi Gallon, and Ádamo Gabriel Cavalcanti Limeira in Brazil.
I wish to express my sincere thanks to them all for their efforts.
I was fortunate to discuss on phylogenetics with Santiago Nava; his advice
was definitive when inferring evolutionary relationships for many results of this study.
He also provided rapid assistance with Principal Component Analyses for sets of
morphological traits from soft tick larvae, presented in the first, second and last
chapter of this work. Thanks are also extended to Arlei Marcili, for his useful advices
on phylogenetic tree-construction and for his devote contribution in the elaboration of
many phylogenetic analyses for different chapters of the current work.
A considerable contribution to the results of the current study was made by
some of my laboratory fellows. I am deeply grateful to Francisco B. Costa, who
taught me the PCR technique, and dedicated great efforts in cellular cultures to
achieve Rickettsia isolations from Chilean ticks. Sincere thanks are also extended to
Amália M. Barbieri, Fernanda Nieri and Yohana Acevedo, who also expended days
in order to successfully isolate Rickettsia from soft ticks.
I wish to thank the technicians and assistant personnel from the laboratory for
their indispensable task in preparing daily-work material, and in maintaining the
cleanliness of the facilities.
Finally, I thank to the “Comisión Nacional de Investigación y Tecnología”
(CONICYT, Chile) for the awarded grant (72140079).
RESUMO

MUÑOZ-LEAL, S. Pesquisa dos agentes Anaplasma, Borrelia, Coxiella,
Ehrlichia, Rickettsia e Hepatozoon em carrapatos (Acari: Ixodoidea: Argasidae,
Ixodidae) do Chile e estudo taxonômico de Ornithodoros capensis sensu lato
(Argasidae) na América do Sul [Study on Anaplasma, Borrelia, Coxiella, Ehrlichia,
Rickettsia and Hepatozoon agents in ticks (Acari: Ixodoidea: Argasidae, Ixodidae)
from Chile, and a taxonomic study on Ornithodoros capensis sensu lato (Acari:
Argasidae) in South America]. 2017. 200 f. Tese (Doutorado em Ciências) –
Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São
Paulo, 2017.

Até 2014, o conhecimento científico sobre a diversidade de Ixodoidea no Chile
estava representado por 19 espécies e apenas agentes infecciosos dos gêneros
Borrelia e Rickettsia haviam sido descritos. O objetivo deste estudo foi o de avaliar a
ocorrência de outros patógenos transmitidos por carrapatos por meio de técnicas
moleculares orientadas para a deteção de Anaplasma, Borrelia, Coxiella, Ehrlichia,
Rickettsia e Hepatozoon. As sequências obtidas foram analisadas filogeneticamente,
identificando-se suas posições em comparação à de organismos de papeis
patogénicos já conhecidos. Como os agentes do gênero Coxiella apresentaram
proximidade filogenética em relação a bactérias congenêricas endosimbiontes, os
dados sobre estas foram utilizados para realizar um estudo taxonômico em
carrapatos do complexo Ornithodoros capensis sensu lato. Em geral, os resultados
confirmam a presença de pelo menos três novas espécies de Borrelia, uma nova
Rickettsia, e três novas espécies de Hepatozoon para a ciência. Rickettsia
amblyommatis, Rickettsia hoogstraalii e Rickettsia lusitaniae foram inseridas como
novos agentes associados a carrapatos no Chile. Embora alguns carrapatos fossem
positivos para a presença de bactérias da família Anaplasmataceae, futuros estudos
devem ser desenvolvidos para confirmar a sua condição especifica, especialmente
através da obtenção de maiores fragmentos do gene codificante para RNA 16S. Os
organismos tipo Coxiella são específicos para cada uma das quatro espécies de
carrapatos do grupo O. capensis analisados neste estudo. Portanto, constituem uma
ferramenta de valor taxonômico para confirmar as identidades e limites genéticos
destes. Finalmente, os resultados deste estudo adicionam pelo menos cinco novas
espécies de carrapatos para a família Argasidae no Chile e apontam a ocorrência de
várias morfotipos de condição incerta que precisam de maiores análises para
esclarecer a com certeza a sua posição taxonômica.