Sebastián Muñoz-Leal

Novel Babesia and Hepatozoon agents infecting non-volant small mammals in the Brazilian Pantanal, with the first record of the tick Ornithodoros guaporensis in Brazil

Taking into account the diversity of small terrestrial mammals of the Pantanal, the present study aimed to verify the occurrence of infection by Ehrlichia spp., Anaplasma spp., Rickettsia spp., Hepatozoon spp., Babesia spp. and parasitism by ticks in non-volant small mammals collected in the Brazilian Pantanal. Samples of blood, liver and spleen were collected from 64 captured animals, 22 marsupials and 42 rodents. Pathogen detection was performed by the use of genus-specific Polymerase Chain Reaction (PCR) assays. Ticks collected from the animals consisted of Amblyomma sculptum and Amblyomma triste nymphs, and Ornithodoros guaporensis larvae. None of the vertebrate samples (blood, liver, or spleen) yielded detectable DNA of Rickettsia spp. or Ehrlichia spp. The blood of the rodent Hylaeamys megacephalus yielded an Anaplasma sp. genotype (partial 16S rRNA gene) 99% similar to multiple Anaplasma spp. genotypes around the world. The blood of three rodents of the species Calomys callosus were positive for a novel Hepatozoon sp. agent, phylogenetically related (18S rDNA gene) to distinct Hepatozoon genotypes that have been detected in rodents from different parts of the world. One marsupial (Monodelphis domestica) and three rodents (Thrichomys pachyurus) were positive to novel piroplasmid genotypes, phylogenetically (18S rDNA gene) related to Theileria bicornis, Cytauxzoon manul, and Cytauxzoon felis. The present study provides the first molecular detection of Hepatozoon sp. and piroplasmids in small mammals in Brazil. Additionally, we expanded the distribution of O. guaporensis to Brazil, since this tick species was previously known to occur only in Bolivia.

A new species of Ornithodoros (Acari: Argasidae), parasite of Microlophus spp. (Reptilia: Tropiduridae) from northern Chile

A new species, Ornithodoros microlophi (Acari: Argasidae), belonging to the subgenus Alectorobius is described from larvae collected on the lizards Microlophus atacamensis (Donoso-Barros, 1966) and Microlophus quadrivittatus (Tschudi, 1845) (Squamata: Tropiduridae) in continental and insular localities from northern Chile. Larvae of O. microlophi can be distinguished from other Neotropical species of the genus Ornithodoros by a combination of the following characters, namely 10 pairs of ventral setae, venter with 6 pairs of sternal setae, dorsal plate pyriform, 19-21 pairs of dorsal setae (typically 20), 13 pairs are dorsolateral and 7 pairs are central, and hypostome with dental formula 4/4 in medial portion and apex pointed. Phylogenetic analysis of 16S rDNA sequences suggests that O. microlophi represents an independent lineage within Neotropical species of the Argasidae.

Rickettsia sp. Strain Atlantic Rainforest Infection in a Patient from a Spotted Fever-Endemic Area in Southern Brazil.

Santa Catarina State in southern Brazil is the state with the second highest number of laboratory-confirmed cases of spotted fever illness in Brazil. However, all these cases were confirmed solely by serological analysis (seroconversion to spotted fever group rickettsiae), which has not allowed identification of the rickettsial agent. Here, a clinical case of spotted fever illness from Santa Catarina is shown by seroconversion and molecular analysis to be caused by Rickettsia sp. strain Atlantic rainforest. This is the third confirmed clinical case due to this emerging rickettsial agent in Brazil. Like the previous two cases, the patient presented an inoculation eschar at the tick bite site. Our molecular diagnosis was performed on DNA extracted from the crust removed from the eschar. These results are supported by previous epidemiological studies in Santa Catarina, which showed that nearly 10% of the most common human-biting ticks were infected by Rickettsia sp. strain Atlantic rainforest.

Description of a new soft tick species (Acari: Argasidae: Ornithodoros) associated with stream-breeding frogs (Anura: Cycloramphidae: Cycloramphus) in Brazil

In this study, we present a morphological description of immature and adult specimens of Ornithodoros saraivai n. sp., a tick associated with the frog Cycloramphus boraceiensis (Cycloramphidae) at São Sebastião island, located in the São Paulo state seaboard, Brazil. While larvae of O. saraivai are ecologically related to Ornithodoros faccinii, another soft tick associated with cycloramphids, the combination of 7 sternal pairs, 16 dorsal pairs, a pyriform dorsal plate and a partially toothed hypostome constitute unique characters of the O. saraivai larvae. One undetermined nymphal instar and adults of O. saraivai are similar to mature specimens of the Ornithodoros talaje species group; however, the O. saraivai specimens can be recognized by the presence of a robust bean-shaped spiracle with a large spiracular plate and more than two long seta in palpal article I. Identical partial sequences of the mitochondrial 16S rDNA gene confirmed the identity for all collected stages and for two cohorts of laboratory-reared larvae of O. saraivai. A Bayesian and Maximum Parsimony inferred phylogenetic trees support the position of O. saraivai in a clade with O. faccinii, suggesting the existence of an Ornithodoros lineage that evolved in association with amphibians.

A novel spotted fever group Rickettsia infecting Amblyomma parvitarsum (Acari: Ixodidae) in highlands of Argentina and Chile

The tick Amblyomma parvitarsum (Acari: Ixodidae) has established populations in Andean and Patagonic environments of South America. For the present study, adults of A. parvitarsum were collected in highland areas (elevation >3500 m) of Argentina and Chile during 2009-2013, and tested by PCR for rickettsial infection in the laboratory, and isolation of rickettsiae in Vero cell culture by the shell vial technique. Overall, 51 (62.2%) out of 82 A. parvitarsum adult ticks were infected by spotted fever group (SFG) rickettsiae, which generated DNA sequences 100% identical to each other, and when submitted to BLAST analysis, they were 99.3% identical to corresponding sequence of the ompA gene of Rickettsia sp. strain Atlantic rainforest. Rickettsiae were successfully isolated in Vero cell culture from two ticks, one from Argentina and one from Chile. DNA extracted from the third passage of the isolates of Argentina and Chile were processed by PCR, resulting in partial sequences for three rickettsial genes (gltA, ompB, ompA). These sequences were concatenated and aligned with rickettsial corresponding sequences available in GenBank. Phylogenetic analysis revealed that the A. pavitarsum rickettsial agent grouped under high bootstrap support in a clade composed by the SFG pathogens R. sibirica, R. africae, R. parkeri, Rickettsia sp. strain Atlantic rainforest, and two unnamed SFG agents of unknown pathogenicty, Rickettsia sp. strain NOD, and Rickettsia sp. strain ApPR. The pathogenic role of this A. parvitarsum rickettsia cannot be discarded, since several species of tick-borne rickettsiae that were considered nonpathogenic for decades are now associated with human infections.

A new species of Ornithodoros (Acari: Argasidae) from desert areas of northern Chile

Ornithodoros atacamensis n. sp. is described from larvae collected on the lizard Liolaemus bisignatus and from free-living adults collected in desert areas from the Pan de Azúcar and Llanos de Challe National Parks, in Northern Chile. Additionally, unengorged larvae were obtained from fertilized females, which laid eggs in the laboratory. Morphological and mitochondrial 16S rDNA sequence analyses were performed in order to compare this new soft tick species with other congeneric Neotropical representatives. Larvae of O. atacamensis are morphologically closely related to Ornithodoros talaje sensu stricto, Ornithodoros puertoricensis, Ornithodoros rioplatensis, Ornithodoros guaporensis and Ornithodoros hasei, all belonging to the O. talaje species group. The larval diagnostic characters for this species are a combination of a large pyriform dorsal plate with a length of approximately 300μm, 17 pairs of dorsal setae with five central pairs, hypostome with apex pointed and dental formula 2/2 in most rows, 3/3 apically, and capsule of the Hallers organ oval in shape without reticulations. Phylogenetic analyses inferred from the mitochondrial 16S rRNA gene and a Principal Component Analysis based on morphometric characters provide additional support to the description of O. atacamensis as an independent lineage within the genus clustering within the O. talaje species group.

The tick Ixodes uriae (Acari: Ixodidae): Hosts, geographical distribution, and vector roles.

The seabird tick Ixodes uriae White 1852, has the most extensive geographical distribution of all tick species, including Afrotropical, Australasian, Nearctic, Neotropical and Palearctic Zoogeographic Regions. Additionally, this tick species parasitizes a wide range of seabirds and constitutes a host for several viral and bacterial agents. Considering the current biological knowledge about this tick species, in this article we list localities, hosts, tick-borne microorganisms and viruses transmitted by I. uriae described in the literature and include new geographical records.

A new flea, Ectinorus (Ectinorus) insignis n. sp. (Siphonaptera, Rhopalopsyllidae, Parapsyllinae), with notes on the subgenus Ectinorus in Chile and comments on unciform sclerotization in the superfamily Malacopsylloidea

A list is provided for the species of Ectinorus sensu stricto from Chile. Ectinorus (Ectinorus) insignis n. sp. is described from Chile: this species is characterized by the male genitalia. In the subgenus Ectinorus, the authors report the presence in Chile of E. pilosus Beaucournu & Carmen Castro, 2002 described from Argentina and E. simonsi (Rothschild, 1904) described from Bolivia but also known from Peru. A female neallotype is designated for E. ineptus Johnson, 1957. “Unciform sclerotization” is noted and illustrated for the first time, in all Malacopsylloidea, and a list is given for all studied species.

Le genre Tunga Jarocki, 1838 (Siphonaptera : Tungidae). I – Taxonomie, phylogénie, écologie, rôle pathogène

Pour la première fois, les 12 espèces actuellement décrites dans le genre Tunga sont étudiées sur le plan de la taxonomie et de la répartition. Divers aspects de leur biologie et leur rôle pathogène sont également envisagés, et en particulier leur phylogénie, leur chorologie, leur phénologie, leur sexe-ratio et leurs dermecos.

Isolation and molecular characterization of a relapsing fever Borrelia recovered from Ornithodoros rudis in Brazil

In South America, early reports from more than 50 years ago incriminated Ornithodoros rudis as vector of Borrelia venezuelensis, an agent of tick-borne relapsing fever (TBRF). Herein we report the rediscovery of O. rudis by means of morphological, biological and molecular analyses, which also comprise the first report of this tick species in Brazil. Phylogenetic analysis using partial fragments of mitochondrial 16S rRNA gene suggested that O. rudis forms a monophyletic group with Ornithodoros erraticus. By using laboratory rodents as hosts, we isolated a relapsing fever Borrelia from an infected O. rudis female. Phylogenetic analysis inferred from the rrs, flaB, and glpQ genes of Borrelia spp. placed the spirochete harbored by O. rudis closely related to Borrelia turicatae. Until further genetic evidence is not obtained we are referring to this O. rudis spirochete as B. venezuelensis. This is the first in vitro isolation of a TBRF Borrelia from South America. The presence of O. rudis in Brazil should not be overlooked, since this tick has been historically implicated in human cases of TBRF in Colombia, Panama, and Venezuela. This study provides new reports of O. rudis and B. venezuelensis after decades of scientific silence on these agents.