Molecular survey of potentially pathogenic microorganisms in ticks collected from coatis (Nasua nasua) in Iguaçu National Park, Atlantic Forest biome, southern Brazil.

Human contact with wild animals in synanthropic habits is often mediated by arthropod vectors such as ticks. This is an important method of spreading infectious agents that pose a risk to human health. Thus, this study aimed to molecularly detect Ehrlichia spp., Anaplasma spp., Borrelia spp., and protozoa of the order Piroplasmida in ticks collected from coatis of Iguaçu National Park (PNI), Paraná, Brazil. This study involved 553 ticks DNA, including Amblyomma spp. larvae, Haemaphysalis juxtakochi nymphs, Amblyomma brasiliense, Amblyomma coelebs, and adults of Amblyomma ovale. The DNA extracted from each sample was subjected to polymerase chain reaction (PCR) targeting the genes 23S rRNA for the Anaplasmataceae family, 16S rRNA for Anaplasma spp., dsb for Ehrlichia spp., flaB, 16S rRNA, hpt, and glpQ for Borrelia spp., and 18S rRNA for Piroplasmid protozoans. DNA from Anaplasma sp. was detected in ticks of the species A. coelebs (4/553); Borrelia sp. DNA was detected in A. coelebs (3/553), A. ovale (1/553), and Amblyomma larvae (1/553); and Theileria sp. was detected in A. coelebs (2/553). All tested samples were negative for Ehrlichia spp. Our study constitutes the newest report in South America of these microorganisms, which remain poorly studied.

Borrelia theileri in Bovine in the northern and southeastern regions of Brazil.

The present study aimed to describe the occurrence of Borrelia spp. in cattle in the states of Minas Gerais and Pará in southeastern and northern Brazil, respectively. Bovine whole blood samples were examined by blood smear and polymerase chain reaction (PCR) to detect the flagellin B (flaB) gene of Borrelia spp. Frequencies of positive animals for Borrelia spp. were 1.52% (2/132) in the municipality of Unaí, Minas Gerais, and 14.2% (2/7) in the municipality of Marabá, Pará. Subsequent genetic sequencing confirmed that the detected spirochetes close to the species B. theileri. In both locations, the animals positive for B. theileri were also highly infested by Rhipicephalus microplus ticks. Despite the low frequency of Borrelia spp., the occurrence of this spirochete indicates that further studies are needed to determine the consequences in cattle herds.

Artificial feeding of Ornithodoros rostratus using a silicone membrane system.

The in vitro feeding of ticks facilitates the conduction of studies involving the intrinsic vector-pathogen relationship, susceptibility tests, and resistance to acaricides, in addition to mimicking the use of experimental hosts. The objective of this study was to establish an in vitro feeding system using silicone membranes to supply various diets to the species Ornithodoros rostratus. Each experimental group included 130 first-instar O. rostratus nymphs. The groups were divided according to the diet provided: citrated rabbit blood, citrated bovine blood, bovine blood with antibiotics, and defibrinated bovine blood. The control group was fed directly on rabbits. Ticks were weighed before and after the feeding and monitored individually according to their biological parameters. The results of the experiment demonstrated that the proposed system was efficient in terms of fixation stimulus and satisfactory in terms of tick engorgement, which would allow the maintenance of O. rostratus colonies by using artificial feeding through silicone membranes. All diets provided were efficient for the maintenance of colonies, but the ticks that received citrated rabbit blood displayed similar biological parameters to those observed under in vivo feeding conditions.

Experimental infection of Monodelphis domestica with Rickettsia parkeri.

This study aimed to evaluate the infection of the marsupial Monodelphis domestica (the gray short-tailed opossum) by Rickettsia parkeri and its role as an amplifier of the bacterium for Amblyomma ticks. Ten M. domestica males were inoculated with phosphate-buffered saline (PBS) and 106 Vero cells infected with R. parkeri. In seven animals, inoculation was intramuscular, and in three intraperitoneal. One male (control) received 1 ml of the same vehicle used for inoculation intraperitoneally. The three animals inoculated intraperitoneally were infested with uninfected A. sculptum larvae and nymphs between the 2nd and 9th day post-infection (DPI). Parasitemia was monitored from the 3rd to 9th DPI by polymerase chain reaction, using primers for 17 kDa and ompA. The animals were also clinically evaluated. Of the animals infected intramuscularly, only one was blood-positive by the 5th DPI. The three animals infected intraperitoneally were blood-positive on the 2nd, 5th, 7th, and 9th DPI. Of the ten pools of recovered engorged ticks, six had positive bands. The kidney, liver, heart, and spleen of an intramuscularly infected animal were also positive. The rectal temperature of the animals tested increased only in the first three DPI. The animals inoculated intraperitoneally showed prostration, bristled hair, and weight loss. The study found that R. parkeri was capable of infecting M. domestica, which developed rickettsemia and caused infection in xenodiagnostic ticks.

Experimental infection of Rickettsia parkeri in the Rhipicephalus microplus tick.

This study aimed to evaluate, by means of artificial feeding, the interaction between a pathogenic rickettsia and the hard tick R. microplus. We used partially engorged females fed on calves free of Rickettsia spp. Group 1 (G1), containing 20 ticks, was fed bovine blood only. Group 2 (G2), containing 20 ticks, was fed blood containing uninfected VERO cells, and group 3 (G3), containing 40 ticks, was fed blood containing VERO cells infected with Rickettsia parkeri. Biological parameters of the non-parasitic phase and a possible bacterial transmission to the tick eggs and to guinea pigs were evaluated. At the end of oviposition, all G3 females were PCR-positive for genes specific for the genus Rickettsia. Although no guinea pigs were infected, the experimental infection of R. microplus by R. parkeri caused a deleterious effect on the oviposition and provided the first report of transovarian transmission of rickettsia in this tick.

Analysis of the risk factors relating to cows' resistance to Rhipicephalus microplus ticks during the peripartum.

This study aimed to evaluate season, breed, number of lactations and milk production as risk factors relating to Rhipicephalus microplus infestation in dairy cows during the peripartum. Eighty-four animals were randomly selected through proportional stratified sampling. All engorged and partially engorged female R. microplus specimens measuring 4.5-8.0 mm were counted during the 5 weeks before calving, calving week and 5 weeks after calving. The peripartum had a significant effect [calving (p < 0.05; prevalence ratio (PR)= 3.12) and post calving (p < 0.05; PR = 2.02)] on R. microplus infestation. Although the average tick count was higher during the rainy season than during the dry season, there was no significant difference (p = 0.055; PR = 0.63). The average R. microplus count was significantly higher (p < 0.05; PR = 2.10) in Bos taurus animals, followed by F1 (p < 0.05; PR = 1.64) and Girolando (p < 0.05; PR = 1.39). The average R. microplus count was significantly higher (p < 0.05; PR = 0.97) in first-lactation animals, followed by those at the second, third and, fourth or subsequent lactation. Milk production showed a negative correlation with R. microplus count, such that high-production animals were significantly (p = 0.003; PR = 2.04) more vulnerable to infestation than were low-production animals. First-lactation and high-production B. taurus animals had greatest vulnerability to R. microplus infestation over the peripartum period, and constituted the at-risk group in the dairy herd.

Influence of the physiological state on infestation by Rhipicephalus microplus in dairy cows.

The purpose of this study was to identify the influence of physiological state, season, breed, number of lactations, dairy productivity, and stocking rate on infestation by Rhipicephalus microplus in dairy cows. Two hundred cows were selected through proportional representative sampling at the Seropédica Experimental Station (Pesagro-Rio), Brazil. Fully or partially engorged R. microplus females measuring between 4.5 and 8.0mm were counted on the right side of each animal. The tick infestation prevalence data were analyzed in relation to the risk factors, using multiple logistic regression. Associations between prevalence and its possible influencing determinants were measured using odds ratios. The average tick count of cows during the peripartum and lactation periods was significantly higher (p<0.05; OR=4.82) than the count in dry cows. Taurine animals showed significantly higher infestation (p<0.05; OR=3.28) than pure zebuine animals. Among cross-bred animals, the infestation was higher (p<0.05) in F1 animals (1/2 taurine×1/2 zebuine) than in Girolando animals (5/8 zebuine×3/8 taurine). Primiparous cows (p<0.05) had significantly heavier infestations than multiparous cows. Dairy production showed a positive correlation with tick infestation (p<0.03; OR=2.94), such that the most productive animals were the most parasitized ones. Animals kept at high densities were significantly more heavily infested (p<0.05, OR=6.32) than animals kept at low density. First-lactation and high-productivity taurine animals were more vulnerable to R. microplus, thus comprising a high-risk group in dairy herds.