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.

Amblyomma scalpturatum Neumann, 1906 (Acari: Ixodidae): confirmation in Acre State, Brazil, and description of parasitism in a human.

Amblyomma scalpturatum is a tick species that is unique to South America. It is commonly associated with the Amazon biome and has been reported in some Brazilian states. This tick species exhibits host specificity: it parasitizes tapirs and suidae. Its role in transmitting pathogens to humans is still unknown. Amblyomma scalpturatum is known to be a human-biting tick; however, there is only one report showing that humans make suitable hosts for this species. The knowledge of tick fauna is lacking in the Acre State. This study collected free-living ticks with the aim of finding new records in Acre State. Collections were carried out in Amazon forest fragments in Rio Branco municipality. An A. scalpturatum specimen was identified and submitted sequencing of the ITS-2 gene. This study presents the first molecular confirmation of A. scalpturatum collected in situ in Acre State, North Region, Brazil. This study also presents the first record of a successfully completed feeding by an A. scalpturatum nymph on a human host in the North region of Brazil. Further investigation is needed to ascertain the complete life cycle of this tick species, its seasonality in the environment, and its relationship to pathogens and competence to transmit them.

A new species of Amblyomma (Acari: Ixodidae) associated with monkeys and passerines of the Atlantic rainforest biome, Southeastern Brazil.

Recent studies have reported several larvae of an unidentified Amblyomma species on passerine birds in Atlantic rainforest fragments in southeastern Brazil. These larvae yielded a unique 16S rRNA haplotype designated as Amblyomma sp. haplotype Nazaré, which showed nucleotide identity levels of 91% to Amblyomma parkeri Fonseca & Aragão, 1952 and 88% to Amblyomma longirostre (Koch, 1844). Herein, we describe Amblyomma sp. haplotype Nazaré as a new species, Amblyomma romarioi n. sp. Martins, Luz & Labruna, through a formal description of the male and female adult stages. Amblyomma romarioi is morphologically and genetically most closely related to A. parkeri, A. longirostre and Amblyomma geayi Neumann, 1899. Among males, the rectangular basis capituli and rounded coxa I spurs separates A. romarioi from A. parkeri, A. longirostre, and A. geayi, which have basis capituli triangular or slightly hexagonal, and pointed coxa I spurs. Among females, the V-shaped genital aperture and coxa I rounded spurs of A. romarioi contrasts to the U-shaped genital aperture and coxa I pointed spurs in A. parkeri, A. longirostre, and A. geayi. Larvae of A. romarioi have been collected on 24 species of passerines. The few records of nymphs and adults were on the black-fronted titi monkey Callicebus nigrifrons (Spix, 1823). The current distribution of A. romarioi is restricted to the Brazilian Atlantic rainforest, southeastern Brazil, in areas with altitude between 363 and 1600 m, within the distribution of C. nigrifrons. We discuss ecological features of Amblyomma romarioi, comparatively to A. parkeri, A. longirostre and A. geayi. The present study increases the Brazilian tick fauna to 74 species.

Amblyomma scalpturatum Neumann, 1906 (Acari: Ixodidae): confirmation in Acre State, Brazil, and description of parasitism in a human / Amblyomma scalpturatum Neumann, 1906 (Acari: Ixodidae): confirmação no Estado do Acre, Brasil, e descrição de parasitismo em um humano

Abstract Amblyomma scalpturatum is a tick species that is unique to South America. It is commonly associated with the Amazon biome and has been reported in some Brazilian states. This tick species exhibits host specificity: it parasitizes tapirs and suidae. Its role in transmitting pathogens to humans is still unknown. Amblyomma scalpturatum is known to be a human-biting tick; however, there is only one report showing that humans make suitable hosts for this species. The knowledge of tick fauna is lacking in the Acre State. This study collected free-living ticks with the aim of finding new records in Acre State. Collections were carried out in Amazon forest fragments in Rio Branco municipality. An A. scalpturatum specimen was identified and submitted sequencing of the ITS-2 gene. This study presents the first molecular confirmation of A. scalpturatum collected in situ in Acre State, North Region, Brazil. This study also presents the first record of a successfully completed feeding by an A. scalpturatum nymph on a human host in the North region of Brazil. Further investigation is needed to ascertain the complete life cycle of this tick species, its seasonality in the environment, and its relationship to pathogens and competence to transmit them.

Resumo Amblyomma scalpturatum é uma espécie de carrapato que ocorre apenas na América do Sul. Está comumente associada ao bioma da Amazônia e tem sido relatada em alguns estados brasileiros. Esta espécie de carrapato apresenta especificidade a hospedeiros: parasita antas e suídeos. Sua competência em transmitir agentes patogênicos a seres humanos é ainda desconhecida. Amblyomma scalpturatum é também conhecido por picar seres humanos; entretanto, há apenas um relato que evidencie que seres humanos sejam capazes de serem hospedeiros adequados. Ainda existem lacunas no conhecimento sobre a ixodofauna no estado do Acre. Neste estudo foram coletados carrapatos de vida livre com o objetivo de encontrar novos registros para o estado do Acre. Coletas de carrapatos foram realizadas em fragmentos de floresta Amazônica no município de Rio Branco. Um exemplar de A. scalpturatum foi identificado e submetido a sequenciamento do gene ITS-2. Este estudo apresenta a primeira confirmação molecular de A. scalpturatum coletado in situ no estado do Acre, região Norte, Brasil. Este estudo também apresenta o primeiro relato de parasitismo completo de uma ninfa de A. scalpturatum em um hospedeiro humano na região Norte do Brasil. Mais investigações são necessárias para elucidar o ciclo de vida completo dessa espécie de carrapato, a sazonalidade de seus estádios no meio ambiente, sua relação a agentes patogênicos e competência em transmiti-los.

Morphological abnormality in larvae of Amblyomma oblongoguttatum (Acari: Ixodidae) / Anormalidade morfológica em larva de Amblyomma oblongoguttatum (Acari: Ixodidae)

Uma larva de Amblyomma oblongoguttatum com anormalidade morfológica foi coletada juntamente com outros espécimes (larvas e fêmeas) dessa espécie e com ninfas de Amblyomma naponense morfologicamente normais em um porco do mato - Pecari tacaju - em área rural do estado de Rondônia/Brasil, durante investigação de ambientes de doenças transmitidas por carrapatos. O espécime morfologicamente anormal apresentou uma bifurcação da região posterior do opistossoma, sem outros caracteres morfológicos alterados. Este é o primeiro registro de anormalidade morfológica no estádio de larva de Amblyomma oblongoguttatum.

Amblyomma oblongoguttatum is a tick that is a parasite of mammals and its preferred hosts are the wild pigs, tapirs and small rodents, being collected also parasitizing domestic dogs and eventually humans. An Amblyomma oblongoguttatum larva with morphological abnormality was collected along with other specimens (larvae and females) of this species and with morphologically normal Amblyomma naponense nymphs in a wild pig - Pecari tacaju - in a rural area of the state of Rondônia / Brazil, during research of tick-borne disease environments. The morphologically abnormal specimen showed a bifurcation of the posterior region of the opisthosoma, without other altered morphological characters. This is the first record of morphological abnormality at the larval stage of Amblyomma oblongoguttatum.(AU)

Morphological abnormality in larvae of Amblyomma oblongoguttatum (Acari: Ixodidae) / Anormalidade morfológica em larva de Amblyomma oblongoguttatum (Acari: Ixodidae)

Uma larva de Amblyomma oblongoguttatum com anormalidade morfológica foi coletada juntamente com outros espécimes (larvas e fêmeas) dessa espécie e com ninfas de Amblyomma naponense morfologicamente normais em um porco do mato - Pecari tacaju - em área rural do estado de Rondônia/Brasil, durante investigação de ambientes de doenças transmitidas por carrapatos. O espécime morfologicamente anormal apresentou uma bifurcação da região posterior do opistossoma, sem outros caracteres morfológicos alterados. Este é o primeiro registro de anormalidade morfológica no estádio de larva de Amblyomma oblongoguttatum.

Amblyomma oblongoguttatum is a tick that is a parasite of mammals and its preferred hosts are the wild pigs, tapirs and small rodents, being collected also parasitizing domestic dogs and eventually humans. An Amblyomma oblongoguttatum larva with morphological abnormality was collected along with other specimens (larvae and females) of this species and with morphologically normal Amblyomma naponense nymphs in a wild pig - Pecari tacaju - in a rural area of the state of Rondônia / Brazil, during research of tick-borne disease environments. The morphologically abnormal specimen showed a bifurcation of the posterior region of the opisthosoma, without other altered morphological characters. This is the first record of morphological abnormality at the larval stage of Amblyomma oblongoguttatum.

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.

Amblyomma ticks infesting amphibians and Squamata reptiles from the lower Amazon region, Brazil.

Amphibians and Squamata reptiles belonging to a zoological collection were screened for ectoparasites, which were removed from the hosts and identified using morphological keys. Descriptive statistics and analysis of the association between the parasite and host characteristics (taxonomic group, capture location and habitat) were done. Among the 1256 animals examined (319 amphibians and 937 reptiles), 86 individuals were parasitized, corresponding to a frequency of 6.9% (6.6% reptiles and 7.5% amphibians). Ticks in the adult and nymph stages were identified to the species level; all of them belonged to the species Amblyomma dissimile. The larvae were identified to the genus level and were all Amblyomma sp. In total 69 larvae, 28 nymphs and eight adults were found. The most parasitized species was the frog Rhinella major: 24 parasitized animals of 65 examined (36.9%). There was a difference (P < 0.001) between parasitism by ticks of the genus Amblyomma with regard to the habitat of capture of the parasitized animal, with a higher parasitism rate in hosts that inhabited open areas as compared to animals ensconced in forest areas and edges of forests. New tick-host associations are given.

Morphodifferentiation of Gené's organ in engorged Amblyomma sculptum Berlese, 1888 female ticks (Acari: Ixodidae).

The Gené's organ (GO) secretes a waxy substance on eggs that reduces water loss and has antimicrobial properties. The current study evaluated morphological and histochemical aspects of GO in Amblyomma sculptum from the period of post-feeding - when ticks detach from the host - to the stage just before oviposition. In this species, GO is composed of a corpus and two pairs of glands, namely, cranial and caudal. Glandular cells are joined laterally by a system of interdigitating membranes with junctional complexes. Histochemistry showed that lipid droplets became more evident as GO developed, while glycogen gradually disappeared, and proteins were detected only near the onset of oviposition. The ultrastructural results revealed a marked distension of the cuticle filled with an amorphous material. Glandular cells showed poor endoplasmatic reticulum, many mitochondria mainly in the basal cell poles and a very developed basal labyrinth. We concluded that the development of GO in A. sculptum ticks was continuous and progressive, and it started after detachment from the host. Additionally, the ultrastructure study suggests that gland cells have an important absorption ability and a low synthetic activity, which indicates that the majority of wax precursors are derived from haemolymph.

Comparative analysis of germ cells and DNA of the genus Amblyomma: adding new data on Amblyomma maculatum and Amblyomma ovale species (Acari: Ixodidae).

Among tick species, members of the subfamily Amblyomminae have received special attention, since they serve as vectors for pathogens such as Rickettsia spp. and display cryptic species complexes that make their taxonomical classification challenging. Amblyomma ovale, Amblyomma maculatum, and other species of the genus Amblyomma have shown a long history of taxonomic controversies. Spermiotaxonomy has proved to be a valuable tool in the solution of systematic conflicts in Metazoa that can aid molecular and external morphological analyses in ticks and, overall, provide more robust analyses and results. With this in mind, this study included histological analyses of the reproductive system of the species A. ovale and A. maculatum, as well as the description of morphohistological characters of the male reproductive system of ticks of the genus Amblyomma, in order to evaluate these characters within the current clustering proposals. In addition, 16S rDNA and COI (mitochondrial) molecular markers were used to study the genetic relationships of the species. The results show that the tick male reproductive system and its germ cells contain useful candidate characters for taxonomical analyses of Ixodida.

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.

Neuronal projections from the Haller's organ and palp sensilla to the synganglion of Amblyomma americanum§.

The present study was conducted to elucidate the neuronal pathways between peripheral olfactory and taste sensilla and the synganglion in an Ixodidae tick species. The tarsus of the front legs (olfactory nerves) and the fourth palpal segment (gustatory nerves) of unfed Amblyomma americanum males and females were excised. A neuronal tracer, dextran tetramethylrhodamine, was used for filling of the sensory neurons. The synganglion preparations were examined using a confocal microscope. Neuronal arborizations from the Haller's organ were confined to the olfactory lobes and the first pedal ganglion. The estimated number of olfactory glomeruli ranged from 16 to 22 per olfactory lobe in the females. The number of glomeruli was not counted in males because they were densely packed. Sensory neurons associated with sensilla at the distal end of the palpal organ projected into the palpal ganglion in the synganglion through the palpal nerve. Gustatory sensory neurons associated with palpal sensilla projected into a commissure with several bulges, which are confined in the palpal ganglion. The findings of distinct projection patterns of sensory neurons associated with the Haller's organ and palpal organ in the lone star tick from this study advanced our knowledge on mechanisms of sensory information processing in ticks.

Neuronal projections from the Haller's organ and palp sensilla to the synganglion of Amblyomma americanum / Projeções neuronais do órgão de Haller e sensila palpal até o singânglio de Amblyomma americanum

The present study was conducted to elucidate the neuronal pathways between peripheral olfactory and taste sensilla and the synganglion in an Ixodidae tick species. The tarsus of the front legs (olfactory nerves) and the fourth palpal segment (gustatory nerves) of unfed Amblyomma americanum males and females were excised. A neuronal tracer, dextran tetramethylrhodamine, was used for filling of the sensory neurons. The synganglion preparations were examined using a confocal microscope. Neuronal arborizations from the Hallers organ were confined to the olfactory lobes and the first pedal ganglion. The estimated number of olfactory glomeruli ranged from 16 to 22 per olfactory lobe in the females. The number of glomeruli was not counted in males because they were densely packed. Sensory neurons associated with sensilla at the distal end of the palpal organ projected into the palpal ganglion in the synganglion through the palpal nerve. Gustatory sensory neurons associated with palpal sensilla projected into a commissure with several bulges, which are confined in the palpal ganglion. The findings of distinct projection patterns of sensory neurons associated with the Hallers organ and palpal organ in the lone star tick from this study advanced our knowledge on mechanisms of sensory information processing in ticks.(AU)

O presente estudo foi conduzido para elucidar a trajetória neuronal, entre as sensilas periféricas olfativas e gustativas e o singânglio, em uma espécie de carrapato Ixodidae. O tarso da primeira pata (nervos olfativos) e o quarto segmento palpal (nervos gustativos) de machos e fêmeas não alimentados de Amblyomma americanum foram excisados. Um traçador neuronal, dextran tetrametilrodamina, foi usado para preenchimento dos neurônios sensoriais. Os singânglios foram examinados através de microscopia confocal. Arborizações neuronais do órgão de Haller foram confinadas nos lobos olfativos e primeiro gânglio pedal. O número estimado de glomérulos olfativos variou de 16 a 22 por lobo olfativo nas fêmeas. Em machos, o número de glomérulos não foi contado, pois eles estavam densamente compactados. Os neurônios sensoriais associados com as sensilas, na porção distal do órgão palpal, projetaram-se no gânglio palpal do singânglio através do nervo palpal. Neurônios sensoriais gustativos associados com a sensila palpal projetaram-se numa comissura onde havia vários bulbos. Os resultados obtidos neste estudo de padrões de projeção distintos de neurônios sensoriais associados com os órgãos de Haller e palpal no carrapato A. americanum avançam nosso conhecimento sobre os mecanismos de processamento da informação sensorial em carrapatos.(AU)

Neuronal projections from the Haller's organ and palp sensilla to the synganglion of Amblyomma americanum§ / Projeções neuronais do órgão de Haller e sensila palpal até o singânglio de Amblyomma americanum

Abstract The present study was conducted to elucidate the neuronal pathways between peripheral olfactory and taste sensilla and the synganglion in an Ixodidae tick species. The tarsus of the front legs (olfactory nerves) and the fourth palpal segment (gustatory nerves) of unfed Amblyomma americanum males and females were excised. A neuronal tracer, dextran tetramethylrhodamine, was used for filling of the sensory neurons. The synganglion preparations were examined using a confocal microscope. Neuronal arborizations from the Haller’s organ were confined to the olfactory lobes and the first pedal ganglion. The estimated number of olfactory glomeruli ranged from 16 to 22 per olfactory lobe in the females. The number of glomeruli was not counted in males because they were densely packed. Sensory neurons associated with sensilla at the distal end of the palpal organ projected into the palpal ganglion in the synganglion through the palpal nerve. Gustatory sensory neurons associated with palpal sensilla projected into a commissure with several bulges, which are confined in the palpal ganglion. The findings of distinct projection patterns of sensory neurons associated with the Haller’s organ and palpal organ in the lone star tick from this study advanced our knowledge on mechanisms of sensory information processing in ticks.

Resumo O presente estudo foi conduzido para elucidar a trajetória neuronal, entre as sensilas periféricas olfativas e gustativas e o singânglio, em uma espécie de carrapato Ixodidae. O tarso da primeira pata (nervos olfativos) e o quarto segmento palpal (nervos gustativos) de machos e fêmeas não alimentados de Amblyomma americanum foram excisados. Um traçador neuronal, dextran tetrametilrodamina, foi usado para preenchimento dos neurônios sensoriais. Os singânglios foram examinados através de microscopia confocal. Arborizações neuronais do órgão de Haller foram confinadas nos lobos olfativos e primeiro gânglio pedal. O número estimado de glomérulos olfativos variou de 16 a 22 por lobo olfativo nas fêmeas. Em machos, o número de glomérulos não foi contado, pois eles estavam densamente compactados. Os neurônios sensoriais associados com as sensilas, na porção distal do órgão palpal, projetaram-se no gânglio palpal do singânglio através do nervo palpal. Neurônios sensoriais gustativos associados com a sensila palpal projetaram-se numa comissura onde havia vários bulbos. Os resultados obtidos neste estudo de padrões de projeção distintos de neurônios sensoriais associados com os órgãos de Haller e palpal no carrapato A. americanum avançam nosso conhecimento sobre os mecanismos de processamento da informação sensorial em carrapatos.

Amblyomma mixtum Koch, 1844 (Acari: Ixodidae): First record confirmation in Colombia using morphological and molecular analyses.

Up to some years ago, the taxon Amblyomma cajennense represented a single tick species in the New World, from southern United States to northern Argentina. Recent studies, based on genetic, reproductive and morphological data reorganized this taxon into a complex of the following 6 valid species: A. cajennense sensu stricto, Amblyomma mixtum, Amblyomma sculptum, Amblyomma interandinum, Amblyomma tonelliae, and Amblyomma patinoi. According to this classification, the A. cajennense complex is currently represented in Colombia by only one species, A. patinoi. Because the Colombian land is surrounded by confirmed records of A. mixtum in Panama and Ecuador, and by A. cajennense s.s. in Venezuela and the Brazilian Amazon, it is possible that these two species could also occur in Colombia. This study aimed to determine the occurrence of ticks of the A. cajennense complex in the Orinoquía region of Colombia. A total of 246 adult ticks of the Amblyomma genus were collected in three sampled regions: 71 females and 110 males in Arauca (Arauca Department), 27 females and 20 males in Nunchía (Casanare Department), and 10 females and 8 males in Yopal (Casanare Department). Based on morphological and molecular analyses, these ticks were identified as A. mixtum. Molecular analyses consisted of DNA sequences of two molecular markers, the nuclear second internal transcribed spacer (ITS2) and the mitochondrial cytochrome c oxidase subunit I gene (COI). The presence of A. mixtum in Colombia is of medical relevance, since this species is incriminated as a vector of Rickettsia rickettsii in Central America.

Geographical distribution of Amblyomma cajennense (sensu lato) ticks (Parasitiformes: Ixodidae) in Brazil, with description of the nymph of A. cajennense (sensu stricto).

BACKGROUND: Until recently, Amblyomma cajennense (Fabricius, 1787) was considered to represent a single tick species in the New World. Recent studies have split this taxon into six species. While the A. cajennense species complex or A. cajennense (sensu lato) (s.l.) is currently represented by two species in Brazil, A. cajennense (sensu stricto) (s.s.) and Amblyomma sculptum Berlese, 1888, their geographical distribution is poorly known. METHODS: The distribution of the A. cajennense (s.l.) in Brazil was determined by morphological examination of all lots of A. cajennense (s.l.) in two large tick collections of Brazil, and by collecting new material during three field expeditions in the possible transition areas between the distribution ranges of A. cajennense (s.s.) and A. sculptum. Phylogenetic analysis inferred from the ITS2 rRNA gene was used to validate morphological results. Morphological description of the nymphal stage of A. cajennense (s.s.) is provided based on laboratory-reared specimens. RESULTS: From the tick collections, a total 12,512 adult ticks were examined and identified as 312 A. cajennense (s.s.), 6,252 A. sculptum and 5,948 A. cajennense (s.l.). A total of 1,746 ticks from 77 localities were collected during field expeditions, and were identified as 249 A. cajennense (s.s.), 443 A. sculptum, and 1,054 A. cajennense (s.l.) [these A. cajennense (s.l.) ticks were considered to be males of either A. cajennense (s.s.) or A. sculptum]. At least 23 localities contained the presence of both A. cajennense (s.s.) and A. sculptum in sympatry. DNA sequences of the ITS2 gene of 50 ticks from 30 localities confirmed the results of the morphological analyses. The nymph of A. cajennense (s.s.) is morphologically very similar to A. sculptum. CONCLUSION: Our results confirmed that A. cajennense (s.l.) is currently represented in Brazil by only two species, A. cajennense (s.s.) and A. sculptum. While these species have distinct distribution areas in the country, they are found in sympatry in some transition areas. The current distribution of A. cajennense (s.l.) has important implications to public health, since in Brazil A. sculptum is the most important vector of the bacterium Rickettsia rickettsii, the etiological agent of Brazilian spotted fever.

Inhibition of the classical pathway of the complement system by saliva of Amblyomma cajennense (Acari: Ixodidae).

Inhibition of the complement system during and after haematophagy is of utmost importance for tick success in feeding and tick development. The role of such inhibition is to minimise damage to the intestinal epithelium as well as avoiding inflammation and opsonisation of salivary molecules at the bite site. Despite its importance, the salivary anti-complement activity has been characterised only in species belonging to the Ixodes ricinus complex which saliva is able to inhibit the alternative and lectin pathways. Little is known about this activity in other species of the Ixodidae family. Thus, the aim of this study was to describe the inhibition of the classical pathway of the complement system by the saliva of Amblyomma cajennense at different stages of the haematophagy. The A. cajennense saliva and salivary gland extract (SGE) were able to inhibit the complement classical pathway through haemolytic assays with higher activity observed when saliva was used. The anti-complement activity is present in the salivary glands of starving females and also in females throughout the whole feeding process, with significant higher activity soon after tick detachment. The SGE activity from both females fed on mice or horses had no significant correlation (p > 0.05) with tick body weight. The pH found in the intestinal lumen of A. cajennense was 8.04 ± 0.08 and haemolytic assays performed at pH 8.0 showed activation of the classical pathway similarly to what occurs at pH 7.4. Consequently, inhibition could be necessary to protect the tick enterocytes. Indeed, the inhibition observed by SGE was higher in pH 8.0 in comparison to pH 7.4 reinforcing the role of saliva in protecting the intestinal cells. Further studies should be carried out in order to identify the inhibitor molecule and characterise its inhibition mechanism.

Amblyomma auricularium (Acari: Ixodidae): underwater survival of the non-parasitic phase of feeding females.

To determine the effects of immersion in water on the biological parameters of engorged females of the tick species Amblyomma auricularium, 60 females were distributed in six groups, each comprising 10 individuals. The control group - G1 (not immersed) was fixed dorsally in a Petri dish and incubated at 27 ± 1°C and 80% RH. The other groups were subjected to immersion periods of 24, 48, 72 and 96 hours, and the sixth group to continuous immersion. After the immersion period, the females were placed in Petri dishes to begin laying. Eggs were collected every 72 hours and kept in biological chambers. All the groups showed significant differences (p <0.05) during the pre-oviposition period. The laying period and the average weight of overall posture did not change. The egg incubation period also did not differ significantly, but the hatching rate in the group immersed for 96h showed a significant difference. Thus, immersion for up to 96 hours does not impair the survival of A. auricularium females, although it may delay egg laying and reduce the number of offspring.

Amblyomma auricularium (Acari: Ixodidae): underwater survival of the non-parasitic phase of feeding females / Amblyomma auricularium (Acari: Ixodidae): sobrevivência subaquática da fase não parasitária de fêmeas alimentadas

To determine the effects of immersion in water on the biological parameters of engorged females of the tick species Amblyomma auricularium, 60 females were distributed in six groups, each comprising 10 individuals. The control group G1 (not immersed) was fixed dorsally in a Petri dish and incubated at 27 ± 1°C and 80% RH. The other groups were subjected to immersion periods of 24, 48, 72 and 96 hours, and the sixth group to continuous immersion. After the immersion period, the females were placed in Petri dishes to begin laying. Eggs were collected every 72 hours and kept in biological chambers. All the groups showed significant differences (p <0.05) during the pre-oviposition period. The laying period and the average weight of overall posture did not change. The egg incubation period also did not differ significantly, but the hatching rate in the group immersed for 96h showed a significant difference. Thus, immersion for up to 96 hours does not impair the survival of A. auricularium females, although it may delay egg laying and reduce the number of offspring.

A fim de conhecer os efeitos da imersão em água sobre os parâmetros biológicos de fêmeas ingurgitadas de Amblyomma auricularium, 60 fêmeas foram distribuídas em seis grupos, cada um contendo 10 indivíduos. O grupo controle G1 (sem imersão) foi fixado dorsalmente numa placa de Petri e incubado a 27 ± 1°C e 80 % de HR. Os demais grupos foram submetidos a períodos de imersão de 24, 48, 72 e 96 horas e, o último grupo, em imersão continua. Após o período de imersão, as fêmeas foram colocadas em placas de Petri para iniciar a postura. Os ovos foram coletados a cada 72 horas e mantidos em câmaras biológicas. Houve diferença significativa (p <0,05) em relação ao período de pré-oviposição de todos os grupos. O período de postura e o peso médio da postura total não se alterou. O período de incubação dos ovos também não diferiram significativamente, mas houve uma diferença significativa na taxa de eclosão das larvas no grupo imerso por 96 horas. Assim, a imersão por até 96 horas não compromete a sobrevivência de fêmeas de A. auricularium, mas pode retardar a postura de ovos e reduzir o número de descendentes.

Amblyomma auricularium (Acari: Ixodidae): underwater survival of the non-parasitic phase of feeding females / Amblyomma auricularium (Acari: Ixodidae): sobrevivência subaquática da fase não parasitária de fêmeas alimentadas

To determine the effects of immersion in water on the biological parameters of engorged females of the tick species Amblyomma auricularium, 60 females were distributed in six groups, each comprising 10 individuals. The control group – G1 (not immersed) was fixed dorsally in a Petri dish and incubated at 27 ± 1°C and 80% RH. The other groups were subjected to immersion periods of 24, 48, 72 and 96 hours, and the sixth group to continuous immersion. After the immersion period, the females were placed in Petri dishes to begin laying. Eggs were collected every 72 hours and kept in biological chambers. All the groups showed significant differences (p <0.05) during the pre-oviposition period. The laying period and the average weight of overall posture did not change. The egg incubation period also did not differ significantly, but the hatching rate in the group immersed for 96h showed a significant difference. Thus, immersion for up to 96 hours does not impair the survival of A. auricularium females, although it may delay egg laying and reduce the number of offspring.

A fim de conhecer os efeitos da imersão em água sobre os parâmetros biológicos de fêmeas ingurgitadas de Amblyomma auricularium, 60 fêmeas foram distribuídas em seis grupos, cada um contendo 10 indivíduos. O grupo controle G1 (sem imersão) foi fixado dorsalmente numa placa de Petri e incubado a 27 ± 1°C e 80 % de HR. Os demais grupos foram submetidos a períodos de imersão de 24, 48, 72 e 96 horas e, o último grupo, em imersão continua. Após o período de imersão, as fêmeas foram colocadas em placas de Petri para iniciar a postura. Os ovos foram coletados a cada 72 horas e mantidos em câmaras biológicas. Houve diferença significativa (p <0,05) em relação ao período de pré-oviposição de todos os grupos. O período de postura e o peso médio da postura total não se alterou. O período de incubação dos ovos também não diferiram significativamente, mas houve uma diferença significativa na taxa de eclosão das larvas no grupo imerso por 96 horas. Assim, a imersão por até 96 horas não compromete a sobrevivência de fêmeas de A. auricularium, mas pode retardar a postura de ovos e reduzir o número de descendentes.