Density and behavior of capybara (Hydrochoerus hydrochaeris) ticks (Acari: Ixodidae) Amblyomma sculptum and Amblyomma dubitatum with notes on Rickettsia bellii infection: Assessing human exposure risk.

In several urban and peri­urban areas of Brazil, populations of Amblyomma sculptum and Amblyomma dubitatum ticks are maintained by capybaras (Hydrochoerus hydrochaeris). In some of these areas, this host and these tick species are associated with Brazilian spotted fever (BSF), a lethal human disease caused by the bacterium Rickettsia rickettsii. In this work, we evaluated the risk of human exposure to these tick species using four collection techniques to discern host-seeking behavior. The study was carried out in 10 urban sites inhabited by capybaras in Uberlândia, a BSF-free municipality in southeastern Brazil. Ticks were collected in areas of 400 m2 at each site and at three seasons. Within the same municipality, the distance and speed of A. sculptum nymphs moving towards the CO2 traps were evaluated. In a sample of ticks Rickettsia DNA was investigated. During the study period, 52,953 ticks were collected. Among these, 83.4 % were A. sculptum (1,523 adults, 10,545 nymphs and 32,104 larvae) and 16.6 % were A. dubitatum (464 adults, 2,153 nymphs and 6,164 larvae). An average annual questing tick density of 4.4/m² was observed, with the highest density recorded at one site in autumn (31.8/m²) and the lowest in summer at another site (0.03/m²). The visual search yielded the highest proportion of A. sculptum larvae, constituting 47 % of the total and 63.6 % of all A. sculptum larvae. In contrast, CO2 traps collected a greater proportion of nymphs and adults of A. sculptum ticks. In the case of A. dubitatum, the CO2 trap was the most efficient technique with 57.7 % of captures of this species, especially of nymphs (94.5 % of captures) and adults (97.8 % of captures). Ticks' ambush height on vegetation (9 to 77 cm), observed by visual search 30 times, yielded a total of 20,771 ticks. Of these, 28 (93 %) were A. sculptum ticks, with only two (7 %) identified as A. dubitatum ticks. Among A. sculptum ticks, the nymph was the most attracted stage to humans and larva in the case of A. dubitatum. Amblyomma sculptum adults and nymphs were significantly more attracted to humans than those of A. dubitatum, but A. dubitatum larvae were significantly more attracted than the same stage of A. sculptum. The maximum distance and speed of horizontal displacement for A. sculptum nymphs were five meters and 2.0 m/h, respectively. The only species of Rickettsia detected in ticks, exclusively in A. dubitatum, was R. bellii. Importantly, it was observed that the higher the proportion of A. sculptum in the community of ticks, the lower the rate of infection of A. dubitatum by R. bellii. In conclusion, host-seeking behavior differed between the two tick species, as well as between stages of the same species. A greater restriction of A. dubitatum ticks to the soil was observed, while larvae and nymphs of A. sculptum dispersed higher in the vegetation. The behavior presented by A. sculptum provides greater opportunities for contact with the hosts, while A. dubitatum depends more on an active search for a host, the hunter behavior. Taken together, these observations show that a human being crossing an area infested with A. sculptum and A. dubitatum ticks will have almost exclusive contact with A. sculptum larvae and/or nymphs. Humans in a stationary position (sitting, lying or immobile) are exposed to both tick species, but they are more attractive to adults and mainly nymphs of A. sculptum compared to the corresponding stages of the tick A. dubitatum. The negative effect of A. sculptum on A. dubitatum infection by R. bellii deserves further studies.

Perception of Ticks and Tick-Borne Diseases Worldwide.

In this comprehensive review study, we addressed the challenge posed by ticks and tick-borne diseases (TBDs) with growing incidence affecting human and animal health worldwide. Data and perspectives were collected from different countries and regions worldwide, including America, Europe, Africa, Asia, and Oceania. The results updated the current situation with ticks and TBD and how it is perceived by society with information bias and gaps. The study reinforces the importance of multidisciplinary and international collaborations to advance in the surveillance, communication and proposed future directions to address these challenges.

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.

Novel genotypes of Hepatozoon spp. in small mammals, Brazil.

BACKGROUND: Small mammals (rodents and marsupials) have been poorly explored for the occurrence of apicomplexan (genus Hepatozoon and genera of the order Piroplasmorida) and Anaplasmataceae agents in Brazil. Thus, this study investigated the occurrence of Hepatozoon spp., Piroplasmorida, and Anaplasmataceae agents in small mammals in seven forest fragments in Brazil. METHODS: During 2015-2018, small mammals were captured in six forest fragments in the State of São Paulo (Cerrado and Atlantic Forest biomes) and one fragment in the State of Mato Grosso do Sul (Pantanal biome). Mammal blood, liver, spleen, and lung samples were tested molecularly for the presence of DNA of Hepatozoon, Piroplasmorida, and Anaplasmataceae agents. RESULTS: A total of 524 mammals were captured, comprising seven species of marsupials, 14 rodents, two carnivores, and one Cingulata. Four novel haplotypes (1, 2, 3, 4) of Hepatozoon spp. were detected in small mammals from different biomes. In São Paulo state, haplotype 1 was detected in rodents from Cerrado and a transition area of Cerrado and Atlantic Forest biomes, whereas haplotype 2 was detected in rodents from the Atlantic Forest biome. On the other hand, haplotypes 3 and 4 were restricted to rodents and marsupials, respectively, from the Pantanal biome of Mato Grosso do Sul. No host species shared more than one haplotype. Despite these distinct geographical and host associations, our phylogenetic analyses indicated that the four Hepatozoon haplotypes belonged to the same clade that contained nearly all haplotypes previously reported on rodents and marsupials, in addition to several reptile-associated haplotypes from different parts of the world. No mammal samples yielded detectable DNA of Piroplasmorida agents. On the other hand, the Anaplasmataceae-targeted polymerase chain reaction (PCR) assay amplified a sequence 100% identical to the Wolbachia pipientis endosymbiont of the rodent filarid Litomosoides galizai. CONCLUSIONS: We report a variety of Hepatozoon haplotypes associated with small mammals in three Brazilian biomes: Cerrado, Atlantic Forest, and Pantanal. Through phylogenetic analyses, the Hepatozoon agents grouped in the rodent-marsupial-reptile large clade of Hepatozoon spp. from the world. The detection of a W. pipientis associated with the rodent filarid L. galizai indicates that the rodent was infected by filarial nematodes.

Morphometric Patterns and Blood Biochemistry of Capybaras (Hydrochoerus hydrochaeris) from Human-Modified Landscapes and Natural Landscapes in Brazil.

The capybara, Hydrochoerus hydrochaeris, is the largest extant rodent of the world. To better understand the correlation between size and body mass, and biochemical parameters of capybaras from areas with different degrees of anthropization (i.e., different food supplies), we sampled free-ranging capybaras from areas of natural landscapes (NLs) and human-modified landscapes (HMLs) in Brazil. Analyses of biometrical and biochemical parameters of capybaras showed that animals from HMLs were heavier (higher body mass) than those from NL, a condition possibly related to fat deposit rather than body length, as indicated by Body Condition Index (BCI) analyses. Biochemical parameters indicated higher serum levels of albumin, creatine kinase, cholesterol, fructosamine and total protein among capybaras from HMLs than from NLs; however, when all adult capybaras were analyzed together only cholesterol and triglycerides were positively correlated with body mass. We propose that the biochemical profile differences between HMLs and NLs are related to the obesity condition of capybaras among HMLs. Considering that heavier animals might live longer and reproduce more often, our results could have important implications in the population dynamics of capybaras among HMLs, where this rodent species is frequently represented by overgrowth populations that generate several levels of conflicts with human beings.

Small mammals, ticks and rickettsiae in natural and human-modified landscapes: Diversity and occurrence of Brazilian spotted fever in Brazil.

We studied communities of small mammals and their ticks in endemic (E) and non-endemic (NE) areas for Brazilian spotted fever (BSF), aiming to infer if diversity parameters of parasites and hosts could be related to occurrence and prevalence of rickettsial infection, especially Rickettsia rickettsii. We compared E and NE areas in human-modified landscapes (HMLs) and natural areas (BIO) with no report of BSF cases. Composition and equitability were important components of diversity explaining differences among areas. The marsupial Didelphis albiventris was dominant in HMLs, but not in natural areas, and this opossum was the main host for the tick Amblyomma sculptum, principal vector of R. rickettsii, especially in E areas. Communities of ticks were dominated by A. sculptum, followed by Amblyomma dubitatum in E areas. In NE areas, this dominance was inverted, with more A. dubitatum than A. sculptum infesting small mammals, but the numbers of ticks were much lower than in E areas. Composition and abundance of ticks in natural areas were very dissimilar from HMLs, with the lowest tick burdens. Didelphis albiventris in E areas presented higher seroprevalence and endpoint titres against R. rickettsii than in other areas. At least three Rickettsia species, non-pathogenic to humans, were detected in natural areas (Rickettsia bellii, Rickettsia amblyommatis and 'Candidatus Rickettsia andeanae'), and only one non-pathogenic species in HMLs (R. bellii). Our results suggest that higher diversity of ticks, hosts and rickettsiae could be relevant factors in buffering the effect in BSF occurrence. Particularly for D. albiventris, its importance has to be quantified in further studies considering the epidemiological scenario of BSF.

Exploring the ecological and evolutionary relationships between Rickettsia and hard ticks in the Neotropical region.

This study addresses a meta-analysis of the distribution of Rickettsia spp. in the Neotropical region, as well as their associations with ticks and vertebrates. A total of 219 published reports on Rickettsia in ticks in the target region were compiled, providing 599 records of 31 species of Rickettsia recorded in 50 species of Ixodidae. The aim is to capture the phylogenetic relationships between rickettsiae and the ticks carrying them in the target region, with a focus on the co-speciation ticks-rickettsiae. We compared the phylogeny of ticks, the records of rickettsiae, the environmental gradients colonized by ticks and the effect of the phylogenetic composition of vertebrates feeding ticks on the detection of Rickettsia in ticks. Results show that differences in rickettsial composition in ticks do not depend on the vertebrate's blood-source. This is the first time this result is demonstrated. This study pinpoints that some Neotropical rickettsial organisms are associated with well-defined phylogenetical clusters of ticks. Secondarily, and probably only in a few cases, rickettsiae share species of phylogenetically distant ticks distributed along a gradient of environmental traits in which the ticks overlap (i.e., the different strains of Rickettsia parkeri sensu lato). We outline the importance of some ticks that share hosts and habitat: these ticks may act as "bridges" for the circulation of rickettsial species. There are also many species of Rickettsia that have been detected so far in only one tick species, pointing to a tight relationship or to the lack of data preventing conclusions about the detection of these bacteria in other ticks. Two species, namely Rickettsia amblyommatis and Rickettsia bellii have been recorded in the majority of ticks in the region (mainly Amblyomma spp.) and seem to be not associated with definite tick species because they may be an essential symbiont of the ticks. We conclude that an adequate analysis of rickettsiae-ticks-habitat is necessary to address the human health issues derived from the infections by rickettsiae.

Genomic Analysis of Novel Poxvirus Brazilian Porcupinepox Virus, Brazil, 2019.

We obtained the complete sequence of a novel poxvirus, tentatively named Brazilian porcupinepox virus, from a wild porcupine (Coendou prehensilis) in Brazil that had skin and internal lesions characteristic of poxvirus infection. The impact of this lethal poxvirus on the survival of this species and its potential zoonotic importance remain to be investigated.

Habitat selection in natural and human-modified landscapes by capybaras (Hydrochoerus hydrochaeris), an important host for Amblyomma sculptum ticks.

Human activities are changing landscape structure and function globally, affecting wildlife space use, and ultimately increasing human-wildlife conflicts and zoonotic disease spread. Capybaras (Hydrochoerus hydrochaeris) are linked to conflicts in human-modified landscapes (e.g. crop damage, vehicle collision), as well as the spread and amplification of Brazilian spotted fever (BSF), the most human-lethal tick-borne disease in the world. Even though it is essential to understand the link between capybaras, ticks and BSF, many knowledge gaps still exist regarding the effects of human disturbance in capybara space use. Here, we analyzed diurnal and nocturnal habitat selection strategies of capybaras across natural and human-modified landscapes using resource selection functions (RSF). Selection for forested habitats was higher across human-modified landscapes, mainly during day- periods, when compared to natural landscapes. Across natural landscapes, capybaras avoided forests during both day- and night periods. Water was consistently selected across both landscapes, during day- and nighttime. Distance to water was also the most important variable in predicting capybara habitat selection across natural landscapes. Capybaras showed slightly higher preferences for areas near grasses/shrubs across natural landscapes, and distance to grasses/shrubs was the most important variable in predicting capybara habitat selection across human-modified landscapes. Our results demonstrate human-driven variation in habitat selection strategies by capybaras. This behavioral adjustment across human-modified landscapes may be related to increases in A. sculptum density, ultimately affecting BSF.

Ornithodoros cerradoensis n. sp. (Acari: Argasidae), a member of the Ornithodoros talaje (Guérin-Méneville, 1849) group, parasite of rodents in the Brazilian Savannah.

Ornithodoros cerradoensis n. sp. is described from field-collected and laboratory reared nymphs, males, females, and larvae parasitizing the rodents Cavia aperea and Thrichomys sp. in the Brazilian Savannah. This new species is morphologically and genetically related with the Ornithodoros talaje group and can be separated from other Neotropical species using the following combination of characters: larva with 18 pairs of setae on dorsum (seven anterolateral, four central and seven posterolateral), hypostome with median dentition 2/2; adults provided with large mammillae; dorsal disks surrounded by bulked marginal ridges delimiting barely pebbled areas; three disks in the anterolateral file, and median disk not merging with the posteromedian file. Feeding assays in the laboratory demonstrated that (1) larvae of O. cerradoensis are slow-feeders (∼6 days), (2) first nymphal instar (N1) molts to second instar (N2) without feeding, and (3) N2 and third nymphal instar (N3) engorge rapidly (minutes). With the exception of Ornithodoros hasei nymphs that depict flattened bodies, O. cerradoensis N1, N2, and N3 highly resemble homologous instars of other species in O. talaje sensu lato, therefore are not suitable for morphological comparisons within the group. In addition to morphological signature of larvae and adults that separate this new species; results of cross-mating attempts between O. cerradoensis and Ornithodoros guaporensis a morphologically and phylogenetically closely related species that also parasitizes rodents in the Brazilian Savannah; a Principal Component Analysis using larval characters; and a phylogenetic analysis using mitochondrial markers, support O. cerradoensis as an independent lineage within the Ornithodorinae.

Author Correction: Clinical and serological evaluation of capybaras (Hydrochoerus hydrochaeris) successively exposed to an Amblyomma sculptum-derived strain of Rickettsia rickettsii.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Clinical and serological evaluation of capybaras (Hydrochoerus hydrochaeris) successively exposed to an Amblyomma sculptum-derived strain of Rickettsia rickettsii.

Brazilian spotted fever (BSF), caused by Rickettsia rickettsii, is the most lethal tick-borne disease in the western hemisphere. In Brazil, Amblyomma sculptum ticks are the main vector. Capybaras (Hydrochoerus hydrochaeris), the largest living rodents of the world (adults weighing up to 100 Kg), have been recognized as amplifying hosts of R. rickettsii for A. sculptum in BSF-endemic areas; i.e., once primarily infected, capybaras develop bacteremia for a few days, when feeding ticks acquire rickettsial infection. We conducted experimental infections of five capybaras with an A. sculptum-derived strain of R. rickettsii and performed clinical and bacteremia evaluation during primary and subsequent infections. Bacteremia was detected in all capybaras during primary infection, but not in subsequent infections. All animals seroconverted to R. rickettsii (titres range: 64-32,768), and remained seropositive throughout the study. Primary infection resulted in clinical spotted fever illness in four capybaras, of which two had a fatal outcome. Subsequent infections in seropositive capybaras resulted in no clinical signs. Capybaras developed a sustained immune response that prevented a second bacteremia. This condition may imply a high reproduction rate of capybaras in BSF-endemic areas, in order to continuously generate capybaras susceptible to bacteremia during primary infection.

Electrocutions in free-living black-tufted marmosets (Callithrix penicillata) in anthropogenic environments in the Federal District and surrounding areas, Brazil.

Shrinking natural habitats exposes some non-human primates to the risk of accidents associated with electrical transmission lines. We examined dead marmosets (Callithrix penicillata) collected in the region from January 2015 to April 2018 to determine the animals' cause of death and for electrocuted animals we examined the locations the animals had died as well as the configuration of the power lines at these sites. We also recorded the sex of the animal, the body region affected, and characteristics of the injuries. We diagnosed electrocutions in 11% (n = 34) of the marmosets studied. Most of the affected animals were male (n = 22) with single or double sites of injury on the limbs. Animals were injured in urban (n = 26) and peri-urban (n = 8) areas on lower-voltage alternate current lines, and we detected no seasonality or hotspots of electrocution. Our findings suggest that movement along transmission lines composed of bundled conductors is a major factor in electrocutions of marmosets in the Federal District and surrounding areas. The planning of electrical power grid infrastructure should consider arboreal primates to prevent electrocutions.

Comparative survival of the engorged stages of Amblyomma dubitatum and Amblyomma sculptum in the laboratory: Implications for Brazilian spotted fever epidemiology.

Brazilian spotted fever (BSF), caused by the bacterium Rickettsia rickettsii, is transmitted to humans mainly by the tick Amblyomma sculptum in southeastern Brazil. In most BSF-endemic areas A. sculptum populations are sustained chiefly by capybaras (Hydrochoerus hydrochaeris), which are also the host of another tick species, Amblyomma dubitatum, not implicated in R. rickettsii transmission. Herein, we evaluated the effects of relative humidity (RH), temperature, and water immersion on the successful development of the engorged stages of A. dubitatum and compared them with recently published data under the same experimental conditions for A. sculptum. We showed that free-living developmental stages (engorged larvae, nymphs and females, and eggs) of A. dubitatum had higher survival rates when these stages were submerged in water for different periods of times (24-72 h). On the other hand, higher survival rates were observed for A. sculptum when ticks were incubated at lower RH values, 65 or 78 %, regardless of summer or winter mean temperatures. These results suggest that A. dubitatum is more adapted to humid habitats and seasonally flooding soils than A. sculptum, whereas this latter species should be more adapted than A. dubitatum to drier habitats. The implications of these results for the spatial distribution of A. dubitatum and A. sculptum, in relation to BSF epidemiology, are discussed.

Ornithodoros cerradoensis n. sp. (Acari: Argasidae), a member of the Ornithodoros talaje (Guérin-Méneville, 1849) group, parasite of rodents in the Brazilian Savannah

Ornithodoros cerradoensis n. sp. is described from field-collected and laboratory reared nymphs, males, females, and larvae parasitizing the rodents Cavia aperea and Thrichomys sp. in the Brazilian Savannah. This new species is morphologically and genetically related with the Ornithodoros talaje group and can be separated from other Neotropical species using the following combination of characters: larva with 18 pairs of setae on dorsum (seven anterolateral, four central and seven posterolateral), hypostome with median dentition 2/2; adults provided with large mammillae; dorsal disks surrounded by bulked marginal ridges delimiting barely pebbled areas; three disks in the anterolateral file, and median disk not merging with the posteromedian file. Feeding assays in the laboratory demonstrated that (1) larvae of O. cerradoensis are slow-feeders (∼6 days), (2) first nymphal instar (N1) molts to second instar (N2) without feeding, and (3) N2 and third nymphal instar (N3) engorge rapidly (minutes). With the exception of Ornithodoros hasei nymphs that depict flattened bodies, O. cerradoensis N1, N2, and N3 highly resemble homologous instars of other species in O. talaje sensu lato, therefore are not suitable for morphological comparisons within the group. In addition to morphological signature of larvae and adults that separate this new species; results of cross-mating attempts between O. cerradoensis and Ornithodoros guaporensis a morphologically and phylogenetically closely related species that also parasitizes rodents in the Brazilian Savannah; a Principal Component Analysis using larval characters; and a phylogenetic analysis using mitochondrial markers, support O. cerradoensis as an independent lineage within the Ornithodorinae.

Epidemiology of capybara-associated Brazilian spotted fever.

BACKGROUND: Brazilian spotted fever (BSF), caused by the bacterium Rickettsia rickettsii, has been associated with the transmission by the tick Amblyomma sculptum, and one of its main hosts, the capybara (Hydrochoerus hydrochaeris). METHODS: During 2015-2019, we captured capybaras and ticks in seven highly anthropic areas of São Paulo state (three endemic and four nonendemic for BSF) and in two natural areas of the Pantanal biome, all with established populations of capybaras. RESULTS: The BSF-endemic areas were characterized by much higher tick burdens on both capybaras and in the environment, when compared to the BSF-nonendemic areas. Only two tick species (A. sculptum and Amblyomma dubitatum) were found in the anthropic areas; however, with a great predominance of A. sculptum (≈90% of all ticks) in the endemic areas, in contrast to a slight predominance of A. dubitatum (≈60%) in the nonendemic areas. Tick species richness was higher in the natural areas, where six species were found, albeit with a predominance of A. sculptum (≈95% of all ticks) and environmental tick burdens much lower than in the anthropic areas. The BSF-endemic areas were characterized by overgrowth populations of A. sculptum that were sustained chiefly by capybaras, and decreased populations of A. dubitatum. In contrast, the BSF-nonendemic areas with landscape similar to the endemic areas differed by having lower tick burdens and a slight predominance of A. dubitatum over A.sculptum, both sustained chiefly by capybaras. While multiple medium- to large-sized mammals have been incriminated as important hosts for A. sculptum in the natural areas, the capybara was the only important host for this tick in the anthropic areas. CONCLUSIONS: The uneven distribution of R. rickettsii infection among A. sculptum populations in highly anthropic areas of São Paulo state could be related to the tick population size and its proportion to sympatric A. dubitatum populations.

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.

The Amblyomma maculatum Koch, 1844 (Acari: Ixodidae) group of ticks: phenotypic plasticity or incipient speciation?

BACKGROUND: The goal of this study was to reassess the taxonomic status of A. maculatum, A. triste and A. tigrinum by phylogenetic analysis of five molecular markers [four mitochondrial: 12S rDNA, 16S rDNA, the control region (DL) and cytochrome c oxidase 1 (cox1), and one nuclear: ribosomal intergenic transcribed spacer 2 (ITS2)]. In addition, the phenotypic diversity of adult ticks identified as A. maculatum and A. triste from geographically distinct populations was thoroughly re-examined. RESULTS: Microscopic examination identified four putative morphotypes distinguishable by disjunct geographical ranges, but very scant fixed characters. Analysis of the separated mitochondrial datasets mostly resulted in conflicting tree topologies. Nuclear gene sequences were almost identical throughout the geographical ranges of the two species, suggesting a very recent, almost explosive radiation of the terminal operational taxonomic units. Analysis of concatenated molecular datasets was more informative and indicated that, although genetically very close to the A. maculatum - A. triste lineage, A. tigrinum was a monophyletic separate entity. Within the A. maculatum - A. triste cluster, three main clades were supported. The two morphotypes, corresponding to the western North American and eastern North American populations, consistently grouped in a single monophyletic clade with many shared mitochondrial sequences among ticks of the two areas. Ticks from the two remaining morphotypes, south-eastern South America and Peruvian, corresponded to two distinct clades. CONCLUSIONS: Given the paucity of morphological characters, the minimal genetic distance separating morphotypes, and more importantly the fact that two morphotypes are genetically indistinguishable, our data suggest that A. maculatum and A. triste should be synonymized and that morphological differences merely reflect very recent local adaptation to distinct environments in taxa that might be undergoing the first steps of speciation but have yet to complete lineage sorting. Nonetheless, future investigations using more sensitive nuclear markers and/or crossbreeding experiments might reveal the occurrence of very rapid speciation events in this group of taxa. Tentative node dating revealed that the A. tigrinum and A. maculatum - A. triste clades split about 2 Mya, while the A. maculatum - A.triste cluster radiated no earlier than 700,000 years ago.

Rhipicephalus sanguineus (Latreille, 1806): Neotype designation, morphological re-description of all parasitic stages and molecular characterization.

The aims of this work were to re-describe all parasitic stages of Rhipicephalus sanguineus sensu stricto, to select and deposit a neotype, and to characterize some of its diagnostic molecular traits. A male of R. sanguineus s.s. collected in Montpellier, France, was designated as neotype. The diagnostic characters unique to the male of R. sanguineus s.s. are: spiracular plate elongated and subtriangular in shape with a dorsal prolongation narrow and usually visible dorsally, with the dorsal prolongation narrower than the width of the adjacent festoon; punctations of the scutum moderate in number and unequal in size; marginal groove conspicuous, deep and punctate; posteromedian groove distinct and elongated, and posterolateral grooves often sub-circular, shorter than posteromedian groove; adanal plates long, wide, and subtriangular in shape, with a clear concavity in its inner margin and posterior margin broadly rounded or truncated; accessory adanal plates with the posterior end pointed, narrower than the width of adjacent festoon. The female of R. sanguineus s.s. can be diagnosed by a combination of broadly U-shaped genital aperture, spiracular plate with a narrow dorsal prolongation visible dorsally, basis capituli hexagonal with broad lateral angles, and scutum barely longer than broad with posterior margin sinuous and punctations moderate in number and unequal in size, larger and more numerous along cervical fields. The nymph has a basis capituli sub-triangular dorsally with lateral angles slightly curved and presence of ventral processes, scutum approximately as long as broad with lateral margins nearly straights, posterior margin broadly rounded, and cervical grooves short and sigmoid in shape extending posteriorly to the level of the eyes. The larva is characterized by basis capituli broader than long with lateral angles short and slightly curved and with posterior margin slightly convex, cervical grooves short, shallow and subparallel, and scutum almost twice broader than long. The phylogenetic analysis of DNA sequences support R. sanguineus s.s. as a well-defined taxon when compared with other species of the R. sanguineus group: R. turanicus s.s., R. camicasi, R. guilhoni, R. sulcatus, R. pusillus, R. rossicus and R. leporis. Molecularly R. sanguineus s.s. also encompasses the so-called "temperate lineage" from the New World (Argentina, southern Brazil, Chile, Uruguay, and USA). The evidence currently available supports the presence of R. sanguineus s.s. in Europe (France, Italy, Spain, Switzerland and Portugal) and America (Argentina, Brazil, Chile, Uruguay and U.S.A.), but further studies are needed to determine the exact geographic range of this taxon.

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.