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BackgroundAmblyomma cajennense F. is one of the best known and studied ticks in the New World because of its very wide distribution, its economical importance as pest of domestic ungulates, and its association with a variety of animal and human pathogens. Recent observations, however, have challenged the taxonomic status of this tick and indicated that intraspecific cryptic speciation might be occurring. In the present study, we investigate the evolutionary and demographic history of this tick and examine its genetic structure based on the analyses of three mitochondrial (12SrDNA, d-loop, and COII) and one nuclear (ITS2) genes. Because A. cajennense is characterized by a typical trans-Amazonian distribution, lineage divergence dating is also performed to establish whether genetic diversity can be linked to dated vicariant events which shaped the topology of the Neotropics.ResultsTotal evidence analyses of the concatenated mtDNA and nuclear + mtDNA datasets resulted in well-resolved and fully congruent reconstructions of the relationships within A. cajennense. The phylogenetic analyses consistently found A. cajennense to be monophyletic and to be separated into six genetic units defined by mutually exclusive haplotype compositions and habitat associations. Also, genetic divergence values showed that these lineages are as distinct from each other as recognized separate species of the same genus. The six clades are deeply split and node dating indicates that they started diverging in the middle-late Miocene.ConclusionsBehavioral differences and the results of laboratory cross-breeding experiments had already indicated that A. cajennense might be a complex of distinct taxonomic units. The combined and congruent mitochondrial and nuclear genetic evidence from this study reveals that A. cajennense is an assembly of six distinct species which have evolved separately from each other since at least 13.2 million years ago (Mya) in the earliest and 3.3 Mya in the latest lineages. The temporal and spatial diversification modes of the six lineages overlap the phylogeographical history of other organisms with similar extant trans-Amazonian distributions and are consistent with the present prevailing hypothesis that Neotropical diversity often finds its origins in the Miocene, after the Andean uplift changed the topology and consequently the climate and ecology of the Neotropics.

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The bacterium Rickettsia rickettsii is the etiological agent of an acute, severe disease called Rocky Mountain spotted fever (RMSF) in the United States or Brazilian spotted fever (BSF) in Brazil. In addition to these two countries, the disease has also been reported to affect humans in Mexico, Costa Rica, Panama, Colombia and Argentina (Paddock et al. 2008). Common clinical signs and symptoms are fever, nausea, vomiting, rash, myalgia, anorexia and headache. The case-fatality rate is 5-10% in the United States and 10-40% in Brazil , Angerami et al. 2006, Chapman et al. 2006.Like humans, dogs are also susceptible to R. rickettsii infection. However, despite the wide distribution of R. rickettsii in the Western Hemisphere, reports of R. rickettsii-induced illness in dogs have been restricted to the United States, where the following clinical abnormalities have been observed: fever, lethargy, anorexia, depression, cutaneous petechiae and echymoses, epistaxis, conjunctivitis, ocular discharge, lymph node enlargement, diarrhea, weight loss, dehydration and central nervous system involvement (mostly paraparesis or tetraparesis, ataxia and vestibular syndrome). Hematological abnormalities include anemia, thrombocytopenia and mild leucopenia at the onset of fever followed by leukocytosis (Keenan et al. 1977a, b, Breitschwerdt et al. 1988, Comer 1991. Case-fatality rates in dogs have been reported to be 3% (Greene 1987), 7% (Greene et al. 1985) and 0% in a more recent study (Gasser et al. 2001).Several serological studies among healthy Brazilian dogs indicate that these animals were infected by R. rickettsii, as shown by high antibody titers to R. rickettsii, at least four-fold higher than titers to other Rickettsia species known to occur in Brazil , Pinter et al. 2008. However, nothing is known about the pathogenicity of South American strains of R. rickettsii for dogs. Since ecological differences and genetic polymorphisms between strains of R. rickettsii from the United States and South America have been reported (Eremeeva et al. 2003, Karpathy et al. 2007, it is not known how pathogenic South American strains of R. rickettsii are for dogs. For this purpose, the present study evaluated experimental infection of dogs with a Brazilian strain of R. rickettsii. MATERIALS AND METHODSEight six-month-old female dogs were used in the study, consisting of five mongrels and three beagles. Dogs were provided by a laboratory animal room, where they were reared with no contact with ticks and under strict sanitary control, being regularly treated with antiInfection of dogs with Rickettsia rickettsii • Eliane M Piranda et al. 697worm medication (Pirantel, Febantel, Praziquantel; Vermivet Plus ® , Biovet, Brazil) and immunized with commercial vaccines for enteroviruses, hepatitis, influenza and leptospirosis (Octa-Cino-Vacin ® , Biovet, Brazil). During the two weeks before starting the experiment, the dogs were clinically healthy and their paired serum samples (14 days interval) were shown to contain no reactive antibodies to eithe...

We evaluated if Rickettsia rickettsii-experimentally infected dogs could serve as amplifier hosts for hipicephalus sanguineus ticks. In addition, we checked if Rh. sanguineus ticks that acquired Ri. rickettsii from dogs could transmit the bacterium to susceptible hosts (vector competence), and if these ticks could maintain the bacterium by transstadial and transovarial transmissions. Uninfected larvae, nymphs, and adults of Rh. sanguineus were allowed to feed upon three groups of dogs: groups 1 (G1) and 2 (G2) composed of Ri. rickettsii-infected dogs, infected intraperitoneally and via tick bites, respectively, and group 3 composed of uninfected dogs. After larval and nymphal feeding on rickettsemic dogs, 7.1-15.2% and 35.8-37.9% of the molted nymphs and adults, respectively, were shown by polymerase chain reaction (PCR) to be infected by Ri. rickettsii, confirming that both G1 and G2 dogs were efficient sources of rickettsial infection (amplifier host), resulting in transstadial transmission of the agent. These infected nymphs and adults successfully transmitted Ri. rickettsii to guinea pigs, confirming vector competence after acquisition of the infection from rickettsemic dogs. Transovarial transmission of Ri. rickettsii was observed in engorged females that had been infected as nymphs by feeding on both G1 and G2 dogs, but not in engorged females that acquired the infection during adult feeding on these same dogs. In the first case, filial infection rates were generally <50%. No tick exposed to G3 dogs was infected by rickettsiae in this study. No substantial mortality difference was observed between Ri. rickettsii-infected tick groups (G1 and G2) and uninfected tick group (G3). Our results indicate that dogs can be amplifier hosts of Ri. rickettsii for Rh. sanguineus, although only a minority of immature ticks (<45%) should become infected. It appears that Rh. sanguineus, in the absence of horizontal transmission, would not maintain Ri. rickettsii through successive generations, possibly because of low filial infection rates.

BackgroundAnaplasma phagocytophilum is an emerging pathogen of humans, dogs and other animals, and it is transmitted by ixodid ticks. The objective of the current study was a) detect A. phagocytophilum in dogs and ixodid ticks using real-time Polymerase Chain Reaction (qPCR); and b) Determine important variables associated to host, environment and potential tick vectors that are related to the presence of A. phagocytophilum in dogs domiciled in Rio de Janeiro, Brazil.MethodsWe tested blood samples from 398 dogs and samples from 235 ticks, including 194 Rhipicephalus sanguineus sensu lato, 15 Amblyomma cajennense, 8 Amblyomma ovale and 18 pools of Amblyomma sp. nymphs. A semi-structured questionnaire was applied by interviewing each dog owner. Deoxyribonucleic acid obtained from ticks and dog buffy coat samples were amplified by qPCR (msp2 gene). The sequencing of 16S rRNA and groESL heat shock operon genes and a phylogenetic analysis was performed. The multiple logistic regression model was created as a function of testing positive dogs for A. phagocytophilum.ResultsAmong the 398 blood samples from dogs, 6.03% were positive for A. phagocytophilum. Anaplasma phagocytophilum was detected in one A. cajennense female tick and in five R. sanguineus sensu lato ticks (four males and one female). The partial sequences of the 16S rRNA, and groESL genes obtained were highly similar to strains of A. phagocytophilum isolated from wild birds from Brazil and human pathogenic strains. The tick species collected in positive dogs were R. sanguineus sensu lato and A. cajennense, with A.cajennense being predominant. Tick infestation history (OR = 2.86, CI = 1.98-14.87), dog size (OR = 2.41, IC: 1.51-12.67), the access to forest areas (OR = 3:51, CI: 1.52-16.32), hygiene conditions of the environment in which the dogs lived (OR = 4.35, CI: 1.86-18.63) and Amblyomma sp. infestation (OR = 6.12; CI: 2.11-28.15) were associated with A. phagocytophilum infection in dogs.ConclusionsThis is the first report of A. phagocytophilum in ixodid ticks from Brazil. The detection of A. phagocitophylum in A. cajennense, an aggressive feeder on a wide variety of hosts, including humans, is considered a public health concern.

This study evaluates the impact of cattle-raising on ticks associated with wildlife in the Pantanal region of Brazil, by trapping free-living ticks using a CO(2) tick trap. The traps were equally distributed in two areas: a 600-ha protected area (PA) and the Nhumirim Ranch (NR). Both areas are contiguous and located at the sub-region of Nhecolândia (18 degrees 59'S; 56 degrees 39'W), Pantanal, Brazil. In each area, four habitat types were selected: cerrado (tropical savanna), forest patches, lakes and soda lakes. The last two are temporary lagoons of fresh and salted water, respectively. A total of 980 nymphs, 613 adults and 13 larvae of ticks Amblyomma were collected in 256 h of collection. In a pen of calves infested by Ornithodoros rostratus 114 specimens were collected, including larvae, nymphs and adults. Adults of A. cajennense and A. parvum were more abundant in the savanna at NR, but not differed among habitat types at PA. There was a tendency of having more nymphs in NR than in PA. The higher number of ticks found in the NR in comparison to the PA is not due to the presence of cattle itself but probably due to introduced dogs and horses, habitat alteration.

In the present study the tick prevalence, mean intensity of infestation and species were recorded on birds captured between January 2009 and December 2010 in the Ecological Station Pirapitinga-ESEC from Minas Gerais State, Brazil. A total of 967 birds, from 15 families and 40 species were captured and 165 (17.1 %) individuals were parasitized by ticks. Of these 160 (97 %) belonged to the order Passeriformes. Five tick species were identified: Amblyomma longirostre (n = 274) was the most common species followed by Amblyomma parvum (n = 43), Amblyomma nodosum (n = 39), Amblyomma ovale (n = 24) and Riphicephalus sanguineus (n = 7). None of 61 unengorged larvae molted to nymph. The mean intensity of infestation was 2.7 ± 2.4 ticks per bird (448 ticks/165 birds) ranging from 1 to 10. Only 19 (11.4 %) birds were infested with one species of tick. The remaining birds were infested by two, three or four species of ticks. Also new hosts for all five ticks were recorded. Only nymphs were recorded throughout the year with two similar peaks during autumn and winter 2009 and 2010.