Detecção e caracterização molecular de agentes Anaplasmataceae, Bartonellaceae, Coxiellaceae, Mycoplasmataceae, Rickettsiaceae, Babesiidae e Theileriidae em javalis de vida livre no Estado de São Paulo, Brasil: Detecção molecular de agentes transmitidos por vetores em javalis (Sus scrofa) e carrapatos associados no Brasil, com evidência de possíveis novos genótipos de Ehrlichia, Anaplasma e Hemoplasma
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Data
2022-03-28
Autores
Santana, Matheus de Souza [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
O presente estudo objetivou investigar, por meio de técnicas moleculares, a ocorrência de agentes Anaplasmataceae, Bartonellaceae, Rickettsiaceae, Mycoplasmataceae, Coxiellaceae e Babesiidae/Theileriidae em amostras de sangue de javalis selvagens e seus respectivos carrapatos amostrados no sudeste do Brasil. Para tal, foram analisadas 67 amostras de sangue e 265 carrapatos (264 Amblyomma sculptum e um A. ovale). Na triagem molecular para agentes Anaplasmataceae por meio de ensaio de PCR baseado no gene 16S rRNA, 5,97% amostras de sangue de javalis e 50,54% de carrapatos foram positivas. Em ensaio de PCR para Ehrlichia spp. baseado no gene dsb, 9,24% dos carrapatos foram positivos. Apesar da baixa ocorrência, um possível novo genótipo 16S rRNA de Anaplasma sp. foi detectado nos javalis amostrados. Com base nas análises filogenéticas baseadas nos genes groEL, gltA, sodB e região ITS (23S-5S), verificou-se que carrapatos A. sculptum e A. ovale coletados de javalis carreiam genótipos de Ehrlichia spp. filogeneticamente associados a E. ewingii, E. ruminantium, e novos genótipos de Ehrlichia previamente detectados em equinos, catetos e carrapatos coletados de onçaspintadas. Na triagem para hemoplasmas por meio de qPCR para o gene 16S rRNA, 88,06% das amostras de sangue e 8,69% de carrapatos mostraram-se positivas. Mycoplasma suis, M. parvum e um possível novo genótipo de hemoplasma foram detectados em javalis na região sudeste do Brasil. Na triagem para Bartonella spp. utilizando qPCR baseada no gene nuoG, 3,8% das amostras de carrapatos mostraram-se positivas. Inferências filogenéticas alocaram quatro sequências nuoG e uma gltA de Bartonella no mesmo clado de Bartonella machadoae. Nenhuma amostra de sangue e carrapato de javalis mostrou-se positiva na qPCR para Coxiella burnetii baseada no gene IS111. Por outro lado, apenas 1,6% dos carrapatos mostraram-se positivos no ensaio de nPCR para piroplasmídeos baseado no gene 18S rRNA. Uma sequência 18S rRNA detectada em pool de ninfas de A. sculptum posicionou-se filogeneticamente próxima às sequências Cytauxzoon felis detectadas em felinos nos Estados Unidos. Rickettsia sp. filogeneticamente associada à R. belli foi detectada em um pool de ninfas de A. sculptum. Trata-se da primeira detecção de hemoplasmas, B. machadoae e Cytauxzoon spp. em A. sculptum. Javalis e carrapatos associados parecem não participar do ciclo epidemiológico de C. burnetii na região estudada. Esta espécie de mamífero invasora pode atuar como potencial dispersor de carrapatos infectados com Ehrlichia spp., Bartonella spp., micoplasmas hemotrópicos e Cytauxzoon, podendo trazer implicações epidemiológicas importantes na transmissão de bartoneloses, erliquioses, hemoplasmoses e cytauxzoonose para seres humanos e animais, mais especificamente para equinos, roedores, suínos e gatos.
The present study aimed to investigate, by molecular techniques, the occurrence of Anaplasmataceae, Bartonellaceae, Rickettsiaceae, Mycoplasmataceae, Coxiellaceae e Babesiidae/Theileriidae in blood samples of free-living wild boars and associated ticks in southeastern Brazil. For this purpose, 67 blood samples and 265 ticks (264 Amblyomma sculptum and one A. ovale) were analyzed. In the screening for Anaplasmataceae agents by a PCR assay based on the 16S rRNA gene, 5.97% blood samples and 50.54% ticks were positive. In the PCR assay for Ehrlichia spp. based on the dsb gene, 9.24% of ticks were positive. Despite the low occurrence, a possible new 16S rRNA genotype of Anaplasma sp. was detected in a wild boar’s blood sample. Based on phylogenetic analyses based on the groEL, gltA, sodB genes and ITS (23S5S) intergenic region, it was found that A. sculptum and A. ovale ticks collected from wild boars carry Ehrlichia genotypes phylogenetically associated with E. ewingii, E. ruminantium, and new Ehrlichia genotypes previously detected in horses, peccaries, and ticks collected from jaguars. In the screening for hemoplasmas by a qPCR based on the 16S rRNA gene, 88.06% of blood samples and 8.69% of ticks were positive. Mycoplasma suis, M. parvum and a possible new hemoplasma genotype were detected in wild boars in southeastern Brazil. In the screening for Bartonella spp. using a nuoG-based qPCR assy, 3.8% of tick samples were positive. Phylogenetic inferences positioned four nuoG and one for gltA Bartonella sequences into the same clade as Bartonella machadoae. No blood or tick samples from wild boars showed to be positive in the qPCR for Coxiella burnetii based on the IS111 gene. On the other hand, only 1.6% of ticks was positive in the nested PCR assay for piroplasmids based on the 18S rRNA gene. An 18S rRNA sequence detected in a pool of A. sculptum nymphs was phylogenetically close to Cytauxzoon felis sequences previoously detected in cats from the United States. Rickettsia sp. closely related to R. belli was detected in a pool of A. sculptum nymphs. This is the first report of hemoplasmas, B. machadoae and Cytauxzoon spp. in A. sculptum. Wild boars and associated ticks do not seem to participate in the epidemiological cycle of C. burnetii in the region studied. This invasive mammal species may act as a potential disperser of ticks infected with Ehrlichia spp., Bartonella spp., hemotropic mycoplasmas, and Cytauxzoon, and may bring important epidemiological implications in the transmission of bartonelosis, ehrlichiosis, hemoplasmosis, and cytauxzoonosis to humans and animals, more specifically to horses, rodents, pigs, and cats.
The present study aimed to investigate, by molecular techniques, the occurrence of Anaplasmataceae, Bartonellaceae, Rickettsiaceae, Mycoplasmataceae, Coxiellaceae e Babesiidae/Theileriidae in blood samples of free-living wild boars and associated ticks in southeastern Brazil. For this purpose, 67 blood samples and 265 ticks (264 Amblyomma sculptum and one A. ovale) were analyzed. In the screening for Anaplasmataceae agents by a PCR assay based on the 16S rRNA gene, 5.97% blood samples and 50.54% ticks were positive. In the PCR assay for Ehrlichia spp. based on the dsb gene, 9.24% of ticks were positive. Despite the low occurrence, a possible new 16S rRNA genotype of Anaplasma sp. was detected in a wild boar’s blood sample. Based on phylogenetic analyses based on the groEL, gltA, sodB genes and ITS (23S5S) intergenic region, it was found that A. sculptum and A. ovale ticks collected from wild boars carry Ehrlichia genotypes phylogenetically associated with E. ewingii, E. ruminantium, and new Ehrlichia genotypes previously detected in horses, peccaries, and ticks collected from jaguars. In the screening for hemoplasmas by a qPCR based on the 16S rRNA gene, 88.06% of blood samples and 8.69% of ticks were positive. Mycoplasma suis, M. parvum and a possible new hemoplasma genotype were detected in wild boars in southeastern Brazil. In the screening for Bartonella spp. using a nuoG-based qPCR assy, 3.8% of tick samples were positive. Phylogenetic inferences positioned four nuoG and one for gltA Bartonella sequences into the same clade as Bartonella machadoae. No blood or tick samples from wild boars showed to be positive in the qPCR for Coxiella burnetii based on the IS111 gene. On the other hand, only 1.6% of ticks was positive in the nested PCR assay for piroplasmids based on the 18S rRNA gene. An 18S rRNA sequence detected in a pool of A. sculptum nymphs was phylogenetically close to Cytauxzoon felis sequences previoously detected in cats from the United States. Rickettsia sp. closely related to R. belli was detected in a pool of A. sculptum nymphs. This is the first report of hemoplasmas, B. machadoae and Cytauxzoon spp. in A. sculptum. Wild boars and associated ticks do not seem to participate in the epidemiological cycle of C. burnetii in the region studied. This invasive mammal species may act as a potential disperser of ticks infected with Ehrlichia spp., Bartonella spp., hemotropic mycoplasmas, and Cytauxzoon, and may bring important epidemiological implications in the transmission of bartonelosis, ehrlichiosis, hemoplasmosis, and cytauxzoonosis to humans and animals, more specifically to horses, rodents, pigs, and cats.
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Palavras-chave
Bartonella sp., Coxiella burnetii, Rickettsia sp., Amblyomma