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Vector competence of the group Rhipicephalus sanguineus ticks from Brazil, Argentina and Uruguay for transmission of the bacterium Ehrlichia canis, causative agent of canine monocytic ehrlichiosis

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Author(s):
Jonas Moraes Filho
Total Authors: 1
Document type: Doctoral Thesis
Press: São Paulo.
Institution: Universidade de São Paulo (USP). Faculdade de Medicina Veterinária e Zootecnia (FMVZ/SBD)
Defense date:
Examining board members:
Marcelo Bahia Labruna; Fabio Gregori; Rosangela Zacarias Machado; Arlei Marcili; Matias Pablo Juan Szabo
Advisor: Marcelo Bahia Labruna
Abstract

Philogenetic studies have shown that in Latin America two different groups of the tick species Rhipicephalus sanguineus. These two groups found apart in two different regions, the Temperate region (Chile, Argentina, Uruguay and Southern Brazil) and the Tropical region (From Southern Brazil up to Mexico). It is likely that those two groups are represented by two different species within R. sanguineus group. Studies on ehrlichiosis (caused by Ehrlichiacanis) in the Latin America point thet E. canis is highly prevalent in the Tropical region whereas is rarely report in Temperate region. The present study tests the hypothesis that the distribution of ehrlichiosis are related to the low vectorial competence of the tick within the taxon group R. sanguineus found in the Temperate area and the high vectorial competence of the tick within the taxon group R. sanguineus found in the Tropical area. The objective of this study was to evaluate comparatively the vectorial competence of four R. sanguineus tick population, where three were collected in Temperate Area, from Argentina, Uruguay and State of Rio Grande do Sul (Brazil) and one from Tropical Area from the State of São Paulo (Brazil). Ticks from all four populations were exposed to E. canins through feeding upon infected dogs in acute disease phase, a control group were set up for each population by feeding larvae and nymphs on non infected dogs. All larvae and nymphs collected in this part of the study were allowed to molt in incubator (27oC,>85%RH) to nymphs and adults, respectively. Part of the ticks were selected to underwent molecular analysis, other part were released on susceptible dogs, in order to determine the vectorial competence for each of the tick populations. From the carried out infestations, samples of blood from the infested dogs were drawn weekly during 60 days. Part of the drawn blood was processed for hemogram that yielded that dogs infested with ticks from the São Paulo population, exposed to E. canis during the nymphal stage, showed important alteration on the number of erytrocytes, hemoglobine, packed cell volume, and platelets underneath the threshold for health dogs. All other dogs showed no important alteration in blood parameters. No fever was detected in any of the dogs. The other part of the weekly drawn blood from each dog was processed by real time PCR targeting E. canis DNA and processed by serology (Indirect immunofluorescence) seeking for anti-E. canis antibodies. Molecular analysis (real time PCR) on ticks were carried out in order to detect the prevalence of infected ticks on each population. Results show that only the ticks from the São Paulo population (Tropical Region) were competent in transmitting E. canis to three susceptible dogs, since material from these three fogs yielded positive in real time PCR and sorology tests. All dogs infested with ticks from the Argentina, Urugay and State of Rio Grande do Sul (Temperate Regions) population have not being infected by E. canis. The molecular tests on the ticks support this find since no tick from Argentina and Uruguay population yielded positive in the real time PCR, whereas about 1% of the nymphs and 28% of the adults of ticks from São Paulo population yielded positive in the real time PCR. The ticks from the State of Rio Grande do Sul population showed 4% of positive for real time PCR among the adult ticks tested, but when these ticks fed on susceptible dogs, they were not competent in transmitting the bacterium, what may shown that these PCR positive ticks might have residual bacterium DNA in the midgut. Analysis on the salivary gland from the adult ticks from all four tick populations that were either incoculates in vitro with E. canis or feeding upon infected dogs, showed positive for E. canis DNA only in the São Paulo population. Samples of ticks collected from different regions of Brazil were analyzed by real time PCR in order to detect E. canis bacterium DNA. There were collected R. sanguineus from the cities of São Paulo, Cuiabá, Campo Grande, Paulo Afonso, Cachoeira do Sul, São Luís, Uberlândia, Bandeirantes, Petrolina, Ji-Paraná; e amostras de Montevidéu (Uruguay) e Rafaela (Argentina). The State of Maranhão showed 7.94% of positive ticks, that being the highest prevalence found. Samples from State of Rio Grande do Sul, Campo Grande City, alongside with cities of Rafaela (Argentina) and Montevidéu (Uruguay), showed none positive tick for E. canins bacterium DNA. All presented data contribute for a better understanding on the absence of canine ehrlichiosis cases in the Temperate Latin America (Southern South America) and therefore support the hypothesis that this fact is related to the low vetorial competence of the ticks within the R. sanguineus taxon founded in this region, on the other hand, ticks within the R. sanguineus taxon found in Tropical Latin America show high vectorial competence in E. canis transmission. (AU)