Amblyomma crassum Robinson, 1926 (Parasitiformes: Ixodidae): morphological and molecular confirmation of females and nymphs, with description of the nymphal stage.

New tick records in the western Brazilian Amazon, with notes on rickettsial infection and molecular evidence for Amblyomma crassum in Brazil.

Dichotomous key to larvae of the genus Amblyomma (Parasitiformes: Ixodidae) in Brazil, with morphological description and redescription of the larval stage of 30 Amblyomma species.

Three species of marsupials from the Amazon region (Marmosa cinerea, Caluromys lanatus, and Didelphis marsupialis) and two from the region of São Paulo (Didelphis marsupialis and Didelphis albiventris) were studied. The G-banding pattern of the species with 2n = 14 (M. cinerea and C. lanatus) was very similar, as well as the pattern of G-bands in the species with 22 chromosomes (Didelphis). All of the autosomes of M. cinerea and D. albiventris have centromeric C-bands and the Y chromosome is totally C-band positive. The long arm of the M. cinerea X chromosome is completely C-band positive except for a negative band close to the centromeric region. In D. albiventris the long arm of the X chromosome is C-band positive except for a negative band close to the telomeric region. In M. cinerea the silver-stained nucleolar organizer regions (Ag-NORs) are found in the acrocentric chromosomes, being located in the telomeric region of one pair and in the centromeric region of the other pair. Caluromys lanatus has centromeric Ag-NORs in one acrocentric and in one submetacentric chromosome pairs. Didelphis marsupialis has three chromosome pairs with telomeric Ag-NORs. In D. albiventris the Ag-NORs are terminal and located in both arms of one pair and in the long arm of two pairs of chromosomes.

In non-mammalian amniotes, the parasphenoid is a midline dermal element with a narrow rostral portion, the cultriform process, linked to the interorbital septum and an expanded distal portion, the alae or wings, forming part of the ventral skull base. In mammals, the parasphenoid has not been found in extant monotremes and only a handful of reports of a reduced parasphenoid (a remnant of the cultriform process) have been made for extant marsupials and placentals. Most reports are in serially-sectioned perinatal specimens where the contrast between the intramembranous origin of the parasphenoid and the overlying endochondral basisphenoid facilitates delimiting the different elements forming the skull base. The only report of a parasphenoid remnant in adult marsupials is in the white-eared opossum, Didelphis albiventris, and it was published more than 100 years ago. Here, we report the results of a survey of 576 specimens of Didelphidae and 115 other Marsupialia in the extant collections of the Section of Mammals, Carnegie Museum of Natural History. We observed what we interpret as a parasphenoid remnant in some juveniles and adults from ten of the 27 didelphid species studied: Didelphis albiventris, Didelphis marsupialis, Didelphis virginiana, Marmosa murina, Monodelphis arlindoi, Monodelphis domestica, Philander opossum, Thylamys elegans, Thylamys pusilla, and Thylamys venustus. This element was variable in its presence within the collection, as well as in its size, form, and position. In our largest specific sample, the Virginia opossum, Didelphis virginiana, a parasphenoid was present in 55% of 238 specimens. It is uncertain if the variable occurrence reflects a true absence of the parasphenoid or its loss during specimen preparation. Outside of Didelphidae, we noted a substantial parasphenoid in the microbiothere Dromiciops gliroides, contributing to a midline septum that partially divides the nasopharynx into two channels, and a probable small one in the macropodid Thylogale sp.In extinct mammals and non-mammalian cynodonts, a midline mesocranial ridge interpreted by prior authors as composed of or including a parasphenoid has a wide distribution, supporting the presence of this structure as primitive for Mammalia. It is suggested here that the Miocene platypus Obdurodon has a well-developed parasphenoid further supporting the presence of a parasphenoid as a plesiomorphic feature for Mammalia that is independently lost in some therians and apparently in extant monotremes.

Habitat associations of opossums and rodents in a lowland forest in French Guiana

Molecular detection and characterization of vector-borne agents in common opossums (Didelphis marsupialis) from northeastern Brazil

The riverine barrier model suggests that rivers play a significant role in separating widespread organisms into isolated populations. In this study, we used a comparative approach to investigate the phylogeography of 6 didelphid marsupial species in central Brazil. Specifically, we evaluate the role of the mid-Araguaia River in differentiating populations and estimate divergence time among lineages to assess the timing of differentiation of these species, using mitochondrial DNA sequence data. The 6 didelphid marsupials revealed different intraspecific genetic patterns and structure. The 3 larger and more generalist species, Didelphis albiventris, Didelphis marsupialis, and Philander opossum, showed connectivity across the Araguaia River. In contrast the genetic structure of the 3 smaller and specialist species, Gracilinanus agilis, Marmosa (Marmosa) murina, and Marmosa (Micoureus) demerarae was shaped by the mid-Araguaia. Moreover, the split of eastern and western bank populations of the 2 latter species is consistent with the age of Araguaia River sediments formation. We hypothesize that the role of the Araguaia as a riverine barrier is linked to the level of ecological specialization among the 6 didelphid species and differences in their ability to cross rivers or disperse through the associated habitat types.

Novel Ehrlichia and Hepatozoon genotypes in white-eared opossums (Didelphis albiventris) and associated ticks from Brazil

The parasite fauna of the gray four-eyed opossum, Philander opossum (Linnaeus, 1758), and the common opossum, Didelphis marsupialis Linnaeus, 1758, in Camp du Tigre, French Guiana, is characterized. Nine species from the gastrointestinal system were recovered from both species, which shared 80% of their parasites. The parasite fauna comprised several monoxenous species (63%) and was dominated by Aspidodera raillieti Travassos, 1914, which exhibited high levels of prevalence and abundance in both communities. Only 2 species (Moennigia sp. and Spirura guianensis) had been recorded in other species of mammals. Both species richness and taxonomic composition at the level of component communities from this locality were compared against 11 communities present in the Virginia ( Didelphis virginiana ), white-bellied (Didelphis albiventris), and common opossum from Argentina, Brazil, Mexico, and the United States. Neither host phylogeny nor taxonomy accounted for statistical differences in species richness. There was no statistical difference among species richness values among the 9 localities studied. Taxonomic similarity was analyzed by means of the Jaccard's similarity index, including all, and only common species (occurring in prevalence >10%). The results suggest that sympatric species of marsupials share more species of parasites than parasite communities occurring in conspecific marsupials from different localities. As a consequence, taxonomic composition of these parasite communities varied depending on the locality. Probably, marsupials of the monophyletic Didelphini offer the same compatibility toward their parasites, by presenting them with similar habitats. Subtle differences in lifestyles of the marsupials may determine the chance of encounter between the symbionts and prevent some parasites from completing their life cycles. Further and more rigorous tests are necessary to determine the roles of encounter and compatibility filters, as well as the role of chance, in the structuring of parasite communities in marsupials.

Atualmente, poucos são os dados biométricos de marsupiais disponíveis no Brasil considerando-se sua riqueza de espécies. Baseando-se nisto, as relações de tamanho corporal de populações naturais do gambá-de-orelha-branca, Didelphis albiventris, e do gambá-de-orelha-preta, D. marsupialis, foram verificadas em dois remanescentes florestais de Curitiba, Sul do Brasil, entre fevereiro de 1995 e fevereiro de 1997. Como resultado, houve uma forte relação entre a erupção de dentes, o peso e a maioria das medidas corporais obtidas de ambas as espécies. Em conseqüência, parâmetros como o comprimento da cabeça podem ser utilizados para estimar os demais parâmetros corporais dos marsupiais estudados. Foi também observado dimorfismo sexual quanto ao tamanho corporal e formato da cabeça para D. marsupialis. Comparativamente, as fêmeas adultas de D. albiventris apresentam maior tamanho que as fêmeas adultas de D. marsupialis, pelo menos no Sul do Brasil. Houve evidências de que alguns Indivíduos podem ter um aumento em seu peso corporal durante o outono, talvez como uma maneira de suportar a estação com menor disponibilidade de alimentos. Por outro lado, D. albiventris armazena gorduras mais freqüentemente, independente da estação do ano. Assim, as estratégias adaptativas de ambas as espécies de marsupiais quanto ao tamanho corporal no Sul do Brasil apresentaram algumas diferenças e provavelmente são resultantes de modos de vida também diferentes, apesar de seu parentesco próximo.

There is limited knowledge about tick diversity in the Amazon region. Here, we survey small terrestrial mammals for tick infestation at the Rio Pardo settlement, Amazonas State, Brazil. Sampling included rainy and dry seasons and four ecotones (primary forest, forest in regeneration, field crops and households). Each animal was inspected for ticks, which, if present, were placed in 70% alcohol and identified. Parasitological indexes were calculated and the presence/absence of ticks on hosts was tested for possible associations with independent variables (ecotone, host sex, host order, host family, host age and season). A total of 208 small mammals were captured, 47 individuals (10 species) in the primary forest, 124 (15 species) in the forest in regeneration, 11 (7 species) in the field crops, and 26 (4 species) in the households. A total of 14 small mammals were infested by ticks (overall prevalence: 6.7%; 95% CI: 3.72 - 11.04%), which consisted of 51 specimens that were identified into four species, as follows: Amblyomma humerale (32 nymphs); Ixodes luciae (6 females); Amblyomma coelebs (1 nymph); and Ornithodoros mimon (1 larva). In addition, 11 larvae were retained as Amblyomma spp. Only host order showed association (P = 0.002) with tick infestation, with marsupials 5.5 times more infested than rodents. Our record of O. mimon on D. marsupialis is the first on this host species, and the first record of a Argasidae tick in the Brazilian state of Amazonas. To the best of our knowledge, this is the first study that actively screened free-living terrestrial small mammals and provided data on prevalence, mean intensity and mean abundance of tick infestations in the Brazilian Amazonas state.

The tick Amblyomma humerale Koch is endemic to South America. All host records refer to the adult stage parasitizing tortoises, mostly yellow-footed tortoise, Geochelone denticulata (L.), and red-footed tortoise, Geochelone carbonaria (Spix). The current study reports the presence of A. humerale in the state of Rondônia, Brazil. A total of 215 adult ticks (201 males, 14 females) was collected from six G denticulata in an Indian reserve and nine Geochelone sp. in rural Monte Negro County, giving an overall mean infestation of 14.3 +/- 12.0 (range: 2-44) ticks per tortoise. Male ticks always outnumbered females on the host and nine tortoises had only male ticks. Male ticks were mostly attached in clusters on the ventral sides of the carapace near the anterior and posterior margins, and more rarely on the outer margin of the plastron. All females were found attached to the tortoise skin, at different sites such as head, neck, shoulders or legs. Male ticks were rarely observed attached to the body skin. Seven engorged nymphs collected on small vertebrates from Monte Negro County molted to adults of A. humerale. This included one nymph each on the seven-colored lizard, Plica plica (L), green tree climber, Plica umbra (L.), and wide-foraging lizard, Kentropyx calcarata Spix,three nymphs on the common opossum, Didelphis marsupialis L., and one nymph on the silky anteater, Cyclopes didactylus L. These constitute the first host records for the immature stages of the tick A. humerale.

Cane toads are highly toxic bufonids invasive in several locations throughout the world. Although physiological changes and effects on native predators for Australian populations have been well documented, Florida populations have received little attention. Cane toads were collected from populations spanning the invaded range in Florida to assess relative toxicity, through measuring morphological changes to parotoid glands, likelihood of secretion, and the marinobufagenin (MBG) content of secretion. We found that residual body indices increased in individuals from higher latitude populations, and relative parotoid gland size increased with increasing toad size. There was no effect of latitude on the allometric relationship between gland size and toad size. We observed an increase in likelihood of secretion by cane toads in the field with increasing latitude. Individuals from southern and northern populations did not vary significantly in the quantity of MBG contained in their secretion. Laboratory‐acclimated cane toads receiving injections of epinephrine were more likely to secrete poison with increasing dose, although there was no difference in likelihood of secretion between southern and northern populations. This suggests that differences between populations in the quantities of epinephrine released in the field, due to altered hypothalamic sensitivity upon disturbance, may be responsible for the latitudinal effects on poison secretion. Our results suggest that altered pressures from northward establishment in Florida have affected sympathetic sensitivity and defensive mechanisms of cane toads, potentially affecting risk to native predators.

Despite the increasingly recognized eco-epidemiological importance of ticks as vectors for numerous zoonotic pathogens in urban areas, data regarding the pathogen diversity and co-infection rates in ticks and wildlife hosts in urban and peri-urban Romania are scanty. We aimed to establish the risk of human exposure to co-infected ticks in Cluj-Napoca, a major city in Romania. DNA was isolated from 151 questing ticks: Ixodes ricinus (n = 95), Haemaphysalis punctata (n = 53), Dermacentor reticulatus (n = 2), and Dermacentor marginatus (n = 1); 222 engorged ticks: I. ricinus (n = 164), I. hexagonus (n = 36), H. punctata (n = 16), H. concinna (n = 6), and 70 tissue samples collected from wildlife hosts during 2018 in five urban, and two peri-urban sites. Using a pre-designed Fluidigm real-time PCR dynamic array, all DNA samples were individually screened for the presence of 44 vector-borne pathogens. Subsequently, conventional PCRs were performed for a selection of samples to allow validation and sequencing. In total, 15 pathogens were identified to species and 6 to genus level. In questing ticks, single infections were more common than co-infections. Seven Borrelia spp. were detected in questing I. ricinus, and three in H. punctata ticks. An overall high prevalence 26.35% (95% CI: 19.46–34.22) and diversity of Borrelia burgdorferi sensu lato was seen in urban questing ticks. Other pathogens of the order Rickettsiales were present with variable prevalence. Co-infections occurred in 27.4% (95% CI: 18.72-37.48) of all infected questing ticks. In engorged ticks the overall Bo. burgdorferi sensu lato prevalence was 35.6% (95% CI: 29.29–42.27), with five species present. Pathogens of the order Rickettsiales were also frequently detected. We report for the first time in Romania the presence of Rickettsia aeschlimannii and Rickettsia felis. Overall, from the infected engorged ticks, 69.2% showcased co-infections. In Ixodes spp., dual co-infections, namely Borrelia spp. and Anaplasma phagocytophilum, and Rickettsia helvetica and A. phagocytophilum were the most prevalent. Given the outcome, we underline the need to establish proper tick-surveillance programs in cities and include co-infections in the management plan of tick-borne diseases in Romania.

In the Brazilian Amazon biome, there has been a rise in human spotted fever cases, but still significant knowledge gaps regarding the diversity and epidemiology of the tick-host-Rickettsia relationship. In the herein study, rickettsiae were investigated in ticks from captured live wild hosts in the Amazon biome by PCR targeting a partial sequence of ompB and gltA genes. All 343 ticks were morphologically identified as belonging to five species of the genus Amblyomma. Amblyomma varium (n = 24, 7%) were collected from a Rhinella marina and a Bradypus tridactylus. Amblyomma geayi (n = 51, 15%) were collected from two Bradypus tridactylus. Amblyomma goeldii (n = 116, 34%) were collected from three Tamandua tetradactyla. Amblyomma dissimile (n = 131, 38%) were collected from two Boa constrictor. Amblyomma humerale (n = 21, 6%) were collected from a Chelonoidis spp. Four A. geayi ticks (one female and three males) collected from a three-toed sloth (B. tridactylus) in 2015 were found to be positive for Rickettsia parkeri strain Atlantic Rainforest. The molecular findings herein have confirmed that R. parkeri spotted fever may occur in the Amazon Rainforest associated with A. geayi, expanding the geographical distribution of the R. parkeri strain to the Central Amazon Rainforest.