Morphological anomalies in hard ticks (Acari: Ixodidae) from Brazil.

Tick abnormalities have been unusual in nature, and they can be divided into local and general. In the present study, external morphological anomalies were described in 31 individual adult ticks of 15 different species of Ixodidae, which were collected on wild hosts (20 ticks), domestic hosts (7 ticks), and in the environment (4 ticks) in 11 states of Brazil from 1998 to 2022. Among the 31 tick specimens, 14 (45%) were categorized as local anomalies, and 17 (55%) as general anomalies. The ticks were taxonomically identified into 14 species of Amblyomma, and one species of Rhipicephalus. Local anomalies included malformations of scutum/alloscutum, ectromely, leg atrophy, and a third ectopic spiracular plate. General anomalies included opisthosoma duplication, no expansion of dorsal alloscutum in engorged females, and gynandromorphism; the latter is described for 13 tick specimens. Morphological anomalies in Amblyomma aureolatum, Amblyomma brasiliense, Amblyomma humerale and Amblyomma longirostre are reported for the first time. Although the results herein expand the list of anomalous tick species in the Neotropics, future studies should be conducted to clarify the origin of these anomalies.

Rickettsia rickettsii (Rickettsiales: Rickettsiaceae) Infecting Amblyomma sculptum (Acari: Ixodidae) Ticks and Capybaras in a Brazilian Spotted Fever-Endemic Area of Brazil.

The bacterium Rickettsia rickettsii is the agent of Brazilian spotted fever (BSF), a highly fatal disease that is transmitted in Brazil mainly by the tick Amblyomma sculptum Berlese, which uses capybaras (Hydrochoerus hydrochaeris Linnaeus) as major hosts. In 2015, we captured nine capybaras in a BSF-endemic area of southeastern Brazil. From each capybara, we collected blood sera that were tested through the immunofluorescence assay using Rickettsia spp. antigens, and A. sculptum ticks, processed for isolation of R. rickettsii through guinea pig inoculation. All capybaras (100%) were seroreactive to Rickettsia spp., with highest titers to R. rickettsii. A total of 166 A. sculptum ticks were macerated and inoculated into nine guinea pigs, from which only one presented high fever and seroconverted to R. rickettsii. Blood from this febrile animal was inoculated into additional guinea pigs (guinea pig passages), which also became febrile and seroconverted, or when euthanized during the fever period, their internal organs (spleen, lung) were shown to contain R. rickettsii DNA. The present rickettsial isolate has been maintained cryopreserved as infected guinea pig organs. There was at least one R. rickettsii-infected tick among the 166 macerated ticks, giving a minimal infection rate of 0.6% (1/166). This infection rate is within the range of previous studies, which reported that only 0.05% to at most 1.28% A. sculptum ticks were infected by R. rickettsii in other BSF-endemic areas. These low infection rates support the low incidence of BSF, despite of A. sculptum being the most frequent human-biting tick in southeastern Brazil.

Comparative Susceptibility of Different Populations of Amblyomma sculptum to Rickettsia rickettsii.

The bacterium Rickettsia rickettsii is the etiological agent of Brazilian spotted fever (BSF), which is transmitted in Brazil mainly by the tick Amblyomma sculptum. Herein, larvae and nymphs of six populations of A. sculptum were exposed to R. rickettsii by feeding on needle-inoculated guinea pigs, and thereafter reared on uninfected guinea pigs or rabbits. Two tick populations were exposed to autochthone R. rickettsii strains, whereas four tick populations were exposed to non-autochthone strains. The six geographically different populations of A. sculptum showed different susceptibilities to R. rickettsii, higher among the two tick populations that were exposed to their autochthone R. rickettsii strain. In addition, higher rates of transovarial transmission of R. rickettsii and vector competence success also included the two tick populations that were exposed to autochthone R. rickettsii strains. These results indicate that the susceptibility of A. sculptum to R. rickettsii varies among different tick populations, with a clear bias for higher susceptibility to an autochthone R. rickettsii strain that has already coevolved with a tick population for some time. Our results demonstrated that the R. rickettsii infection induces higher mortality of engorged larvae and nymphs, and tend to reduce the reproductive fitness of engorged females. All together, these results might explain the low R. rickettsii-infection rates of A. sculptum under natural conditions (usually <1%), and indicate that an A. sculptum population should not be able to sustain a R. rickettsii infection for successive tick generations without the creation of new cohorts of infected ticks via horizontal transmission on vertebrate rickettsemic hosts (amplifying hosts). Finally, despite of the ubiquitous distribution of A. sculptum in southeastern and central-western Brazil, most of the populations of this tick species are devoid of R. rickettsii infection. This scenario might be related to two major factors: (i) insufficient numbers of susceptible amplifying hosts; and (ii) lower susceptibilities of many tick populations. While the first factor has been demonstrated by mathematical models in previous studies, the second is highlighted by the results observed in the present study.

Species richness and seasonal dynamics of ticks with notes on rickettsial infection in a Natural Park of the Cerrado biome in Brazil.

This study evaluated the species richness and seasonal dynamics of ticks and rickettsial agents infecting ticks in the largest natural Reserve of the Cerrado biome of Brazil, the Grande Sertão Veredas National Park. During 2012-2014, a total of 9531 host-seeking ticks were collected by dry ice traps and dragging, whereas 1563 ticks were collected from small mammals, and 1186 ticks from domestic animals. Overall, the following 12 tick species were identified: Amblyomma auricularium, Amblyomma dubitatum, Amblyomma naponense, Amblyomma ovale, Amblyomma parvum, Amblyomma sculptum, Amblyomma tigrinum, Amblyomma triste, Dermacentor nitens, Rhipicephalus microplus, Rhipicephalus sanguineus sensu lato, and Ornithodoros mimon. The three most abundant tick species, A. sculptum, A. parvum, and A. triste, are likely to develop one generation per year, with adults predominating between spring and autumn, and immature ticks during autumn-winter. Small mammals seem to be important hosts for immature stages of A. parvum, and A. triste, but not for A. sculptum. Molecular analyses revealed the presence of the human pathogen Rickettsia parkeri in 10% of the A. triste ticks, whereas two agents of unknown pathogenicity, Rickettsia bellii and 'Candidatus Rickettsia andeanae' were found in 7 and 5%, respectively, of the A. parvum ticks. A fourth rickettsial agent, Rickettsia amblyommatis, was found in a single A. sculptum tick. Several Vero cell-established isolates of R. parkeri and R. bellii were obtained from A. triste and A. parvum, respectively. Serological analyses of small mammals suggest that they have been infected by R. parkeri and R. bellii, possibly via natural infestations by A. triste and A. parvum, respectively. Because the Park has suffered low anthropic alterations, our results should provide baseline data that shall be used for future comparisons with other Cerrado areas with higher degree of anthropic changes.

Rickettsia rickettsii Co-feeding Transmission among Amblyomma aureolatum Ticks.

Amblyomma aureolatum ticks are vectors of Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever in Brazil. Maintenance of R. rickettsii in nature depends on horizontal transmission along tick generations. Although such transmission is known to occur when uninfected and infected ticks feed simultaneously on susceptible animals (co-feeding systemic transmission), we investigated co-feeding nonsystemic transmission, which was based on R. rickettsii-infected and -uninfected A. aureolatum ticks feeding simultaneously on guinea pigs immune to R. rickettsii. Our acquisition and transmission infestations demonstrated that horizontal transmission of R. rickettsii by co-feeding ticks on immune hosts with no systemic infection did not occur when uninfected larvae fed distantly from infected nymphs but did occur in a few cases when uninfected larvae fed side-by-side with infected nymphs, suggesting that they shared the same feeding site. The co-feeding nonsystemic transmission type might have no epidemiologic importance for Rocky Mountain spotted fever.

Comparative biology of the tropical and temperate species of Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) under different laboratory conditions.

Recent studies have shown that the taxon Rhipicephalus sanguineus sensu lato (s.l.) is represented in Latin America by two distinct species, designated as 'tropical species' (distributed from Mexico to Brazil) and 'temperate species' (restricted to the southern cone of South America). Since both tropical and temperate species of R. sanguineus s.l. are parasites primarily of domestic dogs, the reasons for their distinct geographical distribution in South America could be related to particular requirements of abiotic conditions for off-host development. With the purpose to test this hypothesis, this study evaluated the off-host developmental stages (eggs, engorged larvae, nymphs and females) of both tick species simultaneously inside incubators with temperature and photoperiod regimens that simulated the summer and winter conditions of tropical Brazil (where the 'tropical species' occurs) and temperate Brazil (where the 'temperate species' occurs). Results showed that the temperate species had significantly higher survival rates than the tropical species, when engorged ticks (larvae, nymphs and females) and eggs were incubated at lower temperatures simulating winter seasons of many parts of the southern cone of South America, where the temperate species is known to occur. These results suggest that the absence of established populations of the tropical species in temperate areas of South America is related to the low overwinter capacity of the tropical species in those areas. Regarding the temperate species, unfed adults that molted from nymphs under summer conditions of either tropical or temperate Brazil remained dormant, at the state of behavioral diapause for at least 20 weeks. Contrastingly, when engorged nymphs of the temperate species were held at winter conditions for at least 3 months, and then transferred to summer conditions to complete molting, no diapause was observed in adult ticks. These results were corroborated by infestation trials, which showed that diapausing adult ticks took more days to attach to rabbits, and did in lesser numbers, when compared to nondiapausing adult ticks. Contextualization of our results in the current literature suggests that absence of established populations of the temperate species in tropical Brazil is linked to the fact that adult ticks would become inactive (diapause) right after molting from nymphs at any period of the year. On the other hand, absence of established populations of the tropical species in temperate Brazil is linked to the fact that this tick species would not enter diapause, and therefore, could not synchronize its life-cycle to avoid the lethal effects of a more severe winter on its developmental stages. Indeed, such assumptions should be corroborated by additional studies testing different populations of the tropical and temperate species, including more studies under natural conditions.

Isolation of Rickettsia rickettsii from the tick Amblyomma sculptum from a Brazilian spotted fever-endemic area in the Pampulha Lake region, southeastern Brazil.

Brazilian spotted fever (BSF) is a tick-borne disease caused by the bacterium Rickettsia rickettsii, the deadliest spotted fever of the world, transmitted in southeastern Brazil mainly by the tick Amblyomma sculptum, a member of the Amblyomma cajennense species complex. In the present study, over 5000 adults of A. sculptum ticks were collected by dry ice traps in the Municipal Ecological Park, alongside the Pampulha Lake region, a BSF-endemic area of Belo Horizonte city, state of Minas Gerais, southeastern Brazil. Ticks were taken alive to the laboratory, where a sample of 2100 specimens was processed for isolation of R. rickettsii. For this purpose, ticks were macerated and intraperitoneally inoculated into guinea pigs. Only one out of 21 inoculated guinea pigs presented high fever within 21days post inoculation with tick homogenates. This febrile animal was euthanized and its internal organs were macerated and inoculated into additional guinea pigs (guinea pig passage). A spleen sample from a febrile guinea pig was used to inoculate Vero cells, resulting in a successful isolation and in vitro establishment of rickettsiae. Rickettsia-infected Vero cells were used for molecular characterization of the rickettsial isolate through PCR and DNA sequencing of fragments of three rickettsial genes (gltA, ompA, and ompB), which were all 100% identical to corresponding sequences of R. rickettsii from GenBank. The present R. rickettsii isolate was designated as strain Pampulha. A minimal infection rate of 0.05% R. rickettsii-infected ticks was estimated for A. sculptum population of the Pampulha Lake region. Our results, coupled with epidemiological evidences, suggest that R. rickettsii strain Pampulha, isolated from A. sculptum ticks in the present study, is the strain responsible for human clinical cases of BSF in the Pampulha Lake region of Belo Horizonte city.

Different lines of evidence used to delimit species in ticks: A study of the South American populations of Amblyomma parvum (Acari: Ixodidae).

The goal of this work was to combine different lines of evidence besides that of molecular markers to delimit species in ticks when the molecular data are not totally congruent. Two groups (Argentina, Brazil) of South American populations of Amblyomma parvum were compared to test whether the splitting of these two lineages suggested by genetic analyses is complete. Comparative studies of reproductive compatibility, morphological analyses of fixed characters, and comparison of population distributions in spatially defined ecological niches were performed.The morphological comparisons of both discrete and morphometric characters showed no differences among A. parvum ticks from Argentina and Brazil. The intercrosses and backcrosses showed evidence of pre- and post-zygotic compatibility between the two groups. No significant differences in environmental traits were found which would justify the separation of the records of A. parvum in distinct groups. Although the gene flow between the two groups of populations is limited, the absence of reproductive barriers, the lack of significant morphological differences, and the absence of significant differences in the niche preferences indicate that populations of A. parvum from Argentina and Brazil should be treated as a single species. The speciation conjectures suggested by some analyses of mitochondrial DNA sequences were not supported when different lines of evidences were compared.

Comparing feeding and reproductive parameters of Amblyomma parvum tick populations (Acari: Ixodidae) from Brazil and Argentina on various host species.

Amblyomma parvum is a Neotropical tick that is widely spread and a potential vector of pathogens, including Rickettsiae. Genetic differences are remarkable between A. parvum populations from Brazil and Argentina. In this work, feeding and reproduction parameters of A. parvum ticks from these two populations were compared on some key host species to evaluate possible differences in host suitability between them. On the whole parameters of these tick populations were similar when fed on the same host and varied similarly on different host species. Still, bovines were more suitable host for Argentinian larvae than for Brazilian cohorts. It was observed that guinea pigs were the best host A. parvum immatures from both origins, as depicted from higher recovery rate of larvae and heavier engorged nymph weights. Canids and bovids were host species most suitable to adults of both tick populations as shown by the highest number of larvae produced by adult females that engorged on these hosts. Taken together, results showed that in spite of the genetic divergence, A. parvum from Argentina and Brazil have similar biological performance on various host species.

Ticks on birds in a forest fragment of Brazilian cerrado (savanna) in the municipality of Uberlândia, State of Minas Gerais, Brazil.

This is a report of tick species, parasite prevalence and infestation intensity of birds in a forest fragment (18° 56' 57'' S and 48° 12' 14'' W) within the Brazilian cerrado (savanna), in the municipality of Uberlândia, State of Minas Gerais, Brazil. A total of 162 birds from 26 species were captured. One adult tick, 296 larvae and 67 nymphs were found on passerine birds. Of these, it was identified 31 larvae and 27 nymphs of Amblyomma longirostre, 17 nymphs of A. nodosum, one A. cajennense larvae and one male of Rhipicephalus sanguineus. All other ticks were identified as Amblyomma sp. larvae (n = 264) or nymphs (n = 26). Overall tick infestation intensity and prevalence were 4.32 ticks/infested bird and 52%, respectively. Sampling of host-seeking ticks on the ground within the forest during a two-year period showed only five Amblyomma sp. nymphs and one adult male of A. nodosum whereas a search for ticks on domestic animals (cattle, horses and dogs) found Rhipicephalus (Boophilus) microplus ticks. Although identification was possible in only 27% of bird ticks there seemed to be no correlation between environmental and domestic animal and bird infestation. It can be assumed that bird infestation may occur above the ground or at specific sites not sampled.

Ticks on birds in a forest fragment of Brazilian cerrado (savanna) in the municipality of Uberlândia, State of Minas Gerais, Brazil / Carrapatos de aves em um fragmento florestal de Cerrado, município de Uberlândia, Estado de Minas Gerais, Brasil

This is a report of ticks species, parasite prevalence and infestation intensity of birds in a forest fragment (18º 56' 57" S and 48º 12' 14" W) within the Brazilian cerrado (savanna), in the municipality of Uberlândia, State of Minas Gerais, Brazil. A total of 162 birds from 26 species were captured. One adult tick, 296 larvae and 67 nymphs were found on passerine birds. Of these, it was identified 31 larvae and 27 nymphs of Amblyomma longirostre, 17 nymphs of A. nodosum, one A. cajennense larvae and one male of Rhipicephalus sanguineus. All other ticks were identified as Amblyomma sp. larvae (n = 264) or nymphs (n = 26). Overall tick infestation intensity and prevalence were 4.32 ticks/infested bird and 52 percent, respectively. Sampling of host-seeking ticks on the ground within the forest during a two-year period showed only five Amblyomma sp. nymphs and one adult male of A. nodosum whereas a search for ticks on domestic animals (cattle, horses and dogs) found Rhipicephalus (Boophilus) microplus ticks. Although identification was possible in only 27 percent of bird ticks there seemed to be no correlation between environmental and domestic animal and bird infestation. It can be assumed that bird infestation may occur above the ground or at specific sites not sampled.

Neste trabalho, são apresentadas as espécies de carrapatos em aves silvestres, sua prevalência e a intensidade de infestação em um fragmento florestal (18º 56' 57" S e 48º 12' 14" W) do Cerrado, no município de Uberlândia, Estado de Minas Gerais. Foram capturadas 162 aves de 26 espécies, mas apenas as aves Passeriformes estavam infestadas. Nestas, foram encontrados um carrapato adulto, 67 ninfas e 296 larvas. Dentre os carrapatos foi possível a identificação de 31 larvas e 27 ninfas de Amblyomma longirostre, 17 ninfas de A. nodosum, uma larva de A. cajennense e um Rhipicephalus sanguineus macho. Todos os outros carrapatos foram classificados como larvas (n = 264) ou ninfas (n = 26) de Amblyomma sp. A intensidade de infestação e a prevalência de carrapatos foram de 4,32 carrapatos/ave infestada e 52 por cento, respectivamente. Na pesquisa de carrapatos em vida livre sobre a vegetação do fragmento florestal, foram capturados apenas cinco ninfas de Amblyomma sp. e um macho adulto de A. nodosum, por um período de dois anos. Carrapatos da espécie Rhipicephalus (Boophilus) microplus foram encontrados parasitando bovinos, cavalos e cães domésticos da região. Mesmo considerando a identificação de apenas 27 por cento dos carrapatos das aves, parece não haver correlação entre infestação ambiental, nos animais domésticos, com a infestação das aves. Pode-se supor que a infestação das aves ocorra acima do solo ou em locais específicos não pesquisados neste trabalho.