We experimentally infected Amblyomma aureolatum ticks with the bacterium Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever (RMSF). These ticks are a vector for RMSF in Brazil. R. rickettsii was efficiently conserved by both transstadial maintenance and vertical (transovarial) transmission to 100% of the ticks through 4 laboratory generations. However, lower reproductive performance and survival of infected females was attributed to R. rickettsii infection. Therefore, because of the high susceptibility of A. aureolatum ticks to R. rickettsii infection, the deleterious effect that the bacterium causes in these ticks may contribute to the low infection rates (<1%) usually reported among field populations of A. aureolatum ticks in RMSF-endemic areas of Brazil. Because the number of infected ticks would gradually decrease after each generation, it seems unlikely that A. aureolatum ticks could sustain R. rickettsii infection over multiple successive generations solely by vertical transmission.
This study evaluated infection by vector-borne agents in 58 crab-eating fox (Cerdocyon thous L.) that were road-killed in an Atlantic rainforest reserve in the state of Espírito Santo, southeastern Brazil. Spleen, lung, or blood samples collected from the foxes were tested in the laboratory by a battery of polymerase chain reaction (PCR) assays targeting bacteria of the genera Rickettsia, Borrelia, Coxiella, Anaplasma, and Ehrlichia; and protozoa of the genera Babesia, Hepatozoon, and Leishmania. Of the targeted organisms, evidence of infection in the foxes was detected for Ehrlichia and Hepatozoon organisms only. Overall, six (10.3%) foxes were infected by an ehrlichial agent closely related to an ehrlichial agent recently detected in free-ranging Jaguars [(Panthera onca (L.)] in central-western Brazil, and to Ehrlichia ruminantium. For Hepatozoon, 28 (48.3%) foxes were infected by an agent closely related to Hepatozoon sp. Curupira 2 and H. americanum; and one (1.7%) fox was infected by an organism closely related to reptile-associated Hepatozoon agents. Finally, 11 (19.0%) foxes were found infested by Amblyomma cajennense (F.) nymphs, which were all PCR negative for the range of vector-borne agents cited above. Because the haplotypes found in free-ranging foxes are genetically closely related to pathogens of great veterinary importance, namely E. ruminantium and H. americanum, it is highly desirable to know if these novel organisms have any important role as agents of diseases in domestic animals and wildlife in Brazil.
Tick-borne bacteria were investigated in 10 free-living jaguars and their ticks in the Pantanal biome, Brazil. Jaguar sera were tested by indirect fluorescent antibody assays using Rickettsia rickettsii, Rickettsia parkeri, Rickettsia amblyommii, Rickettsia rhipicephali, Rickettsia felis, Rickettsia bellii, Ehrlichia canis, and Coxiella burnetii as crude antigens. All 10 jaguar sera reacted (titer ≥ 64) to at least one Rickettsia species; 4 and 3 sera reacted with E. canis and C. burnetii, respectively. One jaguar presented antibody titer to R. parkeri at least fourfold higher than those to any of the other five Rickettsia antigens, suggesting that this animal was infected by R. parkeri. Ticks collected from jaguars included the species Amblyomma cajennense, Amblyomma triste, and Rhipicephalus (Boophilus) microplus. No Rickettsia DNA was detected in jaguar blood samples, but an A. triste specimen collected on a jaguar was shown by PCR to be infected by R. parkeri. The blood of two jaguars and samples of A. triste, A. cajennense, and Amblyomma sp. yielded Ehrlichia DNA by PCR targeting the ehrlichial genes 16S rRNA and dsb. Partial DNA sequences obtained from PCR products resulted in a new ehrlichial strain, here designated as Ehrlichia sp. strain Jaguar. A partial DNA sequence of the 16S rRNA gene of this novel strain showed to be closest (99.0%) to uncultured strains of Ehrlichia sp. from Japan and Russia and 98.7% identical to different strains of Ehrlichia ruminantium. The ehrlichial dsb partial sequence of strain jaguar showed to be at most 80.7% identical to any Ehrlichia species or genotype available in GenBank. Through phylogenetic analysis, Ehrlichia sp. strain jaguar grouped in a cluster, albeit distantly, with different genotypes of E. ruminantium. Results highlight risks for human and animal health, considering that cattle ranching and ecotourism are major economic activities in the Pantanal region of Brazil.
The Borrelia burgdorferi sensu lato (s.l.) group comprises genetically related spirochetes, mostly associated with tick species belonging to the Ixodes ricinus complex in the Northern Hemisphere. The present study evaluated borrelial infection in the tick Ixodes pararicinus, which is the only representative species of the I. ricinus complex in Uruguay. A total of 137 I. pararicinus ticks were collected from deer, cattle, or vegetation in 2 Uruguayan Departments. A part of these ticks was tested directly by PCR targeting the borrelial gene flagellin (fla), whereas another part of the ticks was inoculated into Barbour-Stoenner-Kelly (BSK)-H medium in an attempt to isolate Borrelia. Overall, Borrelia infection was detected in 9 males and 1 nymphal tick pool. These ticks were found to be infected by unique fla haplotypes, which were shown through phylogenetic analysis to represent possibly 2 new B. burgdorferi s.l. genospecies, 1 associated with B. bissettii, the other phylogenetically closest to B. americana. These results were reinforced by PCR and DNA sequencing analyses of portions of 2 additional borrelial genes, rrfA-rrlB intergenic spacer region (IGS) and 16S rDNA (rrs). Weekly examinations of BSK cultures by dark-field microscopy failed to demonstrate live Borrelia through a 100-day incubation period. However, Borrelia DNA was detected by fla-PCR in culture media from 2 vials up to 90 days after inoculation. To the best of our knowledge, this is the first report of B. burgdorferi s.l. infecting ticks in South America.
Mineral and organic additions are often used in mortars to improve their properties. Microstructural investigation concerning the effects of styrene acrylic polymer and silica fume on the mineralogical composition of high-early-strength portland cement pastes after 28 days of hydration are presented in this paper. Thermogravimetry and derivative thermogravimetry were used to study the interaction between polymers and cement, as well as the extent of pozzolanic reaction of the mortars with silica fume. Differential scanning calorimetry and X ray diffraction were used to investigate the cement hydration and the effect of the additions. The results showed that the addition of silica fume and polymer reduces the portlandite formation due to delaying of Portland cement hydration and pozzolanic reaction
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