New records of soft ticks (Acari: Argasidae) from caves in Brazil, with a morphological study of Ornithodoros fonsecai and an analysis of the taxonomic status of Antricola inexpectata.

In this study, we report soft ticks from bat-inhabiting caves in different areas of Brazil. From 2010 to 2019, we collected 807 tick specimens from nine caves located in four Brazilian states among two biomes. Ticks were morphologically identified as Antricola guglielmonei (282 specimens), Ornithodoros cavernicolous (260 specimens), and Ornithodoros fonsecai (265 specimens). Whereas A. guglielmonei was collected on bat guano in hot caves, O. cavernicolous and O. fonsecai were collected in cracks and crevices on the walls of cold caves, sometimes in the same chamber. Morphological identifications were corroborated by molecular and phylogenetic analyses inferred from tick mitochondrial 16S rRNA gene partial sequences. The sequences of A. guglielmonei, O. cavernicolous and O. fonsecai collected in this study clustered with conspecific GenBank sequences from different localities of Brazil. Remarkably, a clade containing 12 sequences of O. fonsecai was clearly bifurcated, denoting a degree of genetic divergence (up to 5 %) of specimens from Cerrado/Atlantic Forest biomes with the specimens from the Caatinga biome. To further evaluate this divergence, we performed morphometric analysis of the larval stage of different O. fonsencai populations by principal component analysis, which indicated that the larvae from Caatinga populations were generally smaller than the larvae from other biomes. Some of the present A. guglielmonei specimens were collected from the type locality of Antricola inexpectata. Comparisons of these specimens with the type specimens of A. inexpectata and A. guglielmonei indicated that they could not be separated by their external morphology. Hence, we are relegating A. inexpectata to a synonym of A. guglielmonei. This proposal is corroborated by our phylogenetic analysis.

New records and phylogenetic position of Ornithodoros knoxjonesi (Ixodida: Argasidae).

Larvae of Ornithodoros knoxjonesi collected at five localities in three countries were studied using morphological and molecular methods to confirm this species' taxonomic validity. The larva of O. knoxjonesi is characterized as having 14 pairs of dorsal setae, eight pairs of ventral setae, plus a posteromedian seta; an elongate dorsal plate, tapered anteriorly; and a hypostome that is narrower near its midlength, with posteriorly projecting denticles. Although the larvae of O. knoxjonesi and Ornithodoros peropteryx are morphologically quite similar, the larva of O. knoxjonesi is characterized as having dorsal setae that are wider at the tip than at the base, while in O. peropteryx these setae are narrower at the tip than at the base; moreover, the dorsal setae are shorter in O. knoxjonesi (Al 0.037-0.065; Pl 0.035-0.059) than in O. peropteryx (Al 0.120-0.132; Pl 0.080-0.096). These species also differ in that O. knoxjonesi possesses only the Al seta on tarsus I, whereas O. peropteryx has both Al and Pl setae. And while both species have two setae on coxae I-III, in O. knoxjonesi the anterior seta is tapering and smooth and the posterior is fringed, while both setae are fringed in O. peropteryx. At the molecular level, based on a maximum likelihood analysis using approximately 400 bp of the mitochondrial 16S rDNA gene, O. knoxjonesi appears as an independent lineage, separated from O. peropteryx by a genetic distance of 16.28 %. Balantiopteryx plicata is a common host of O. knoxjonesi; however, in this work we report Pteronotus personatus and Pteronotus gymnonotus as new hosts of this tick species.

Ornithodoros dyeri (Parasitiformes: Ixodida: Argasidae) parasitizing Leptonycteris yerbabuenae (Chiroptera: Phyllostomidae) in Mexico.

Parasites are a selective force that shape communities and ecosystems. Hosts represent a food source, habitat, and a way to disperse. In recent years, investigations dealing with bats and their role as hosts to numerous parasitic organisms, including metazoan ectoparasites and endoparasites have increased, and soft ticks (Parasitiformes: Ixodida: Argasidae) are among the best known. In Mexico, 16 species of soft ticks associated with bats have been reported up to now, but there are no specific records of soft ticks parasitizing Leptonycteris yerbabuenae in the country. Herein, we record for the first time the presence of Ornithodoros dyeri parasitizing L. yerbabuenae and report an extension of its geographic distribution. The same tick species was also recorded from the California leaf-nosed bat (Macrotus californicus). Data on prevalence, mean intensity, mean abundance, tick identification, and possible scenarios of life cycle associations for O. dyeri and one of its hosts, L. yerbabuenae, are discussed.

The Nothoaspis amazoniensis Complete Mitogenome: A Comparative and Phylogenetic Analysis.

The molecular biology era, together with morphology, molecular phylogenetics, bioinformatics, and high-throughput sequencing technologies, improved the taxonomic identification of Argasidae family members, especially when considering specimens at different development stages, which remains a great difficulty for acarologists. These tools could provide important data and insights on the history and evolutionary relationships of argasids. To better understand these relationships, we sequenced and assembled the first complete mitochondrial genome of Nothoaspis amazoniensis. We used phylogenomics to identify the evolutionary history of this species of tick, comparing the data obtained with 26 complete mitochondrial sequences available in biological databases. The results demonstrated the absence of genetic rearrangements, high similarity and identity, and a close organizational link between the mitogenomes of N. amazoniensis and other argasids analyzed. In addition, the mitogenome had a monophyletic cladistic taxonomic arrangement, encompassed by representatives of the Afrotropical and Neotropical regions, with specific parasitism in bats, which may be indicative of an evolutionary process of cospeciation between vectors and the host.

Molecular evidence and additional morphological characters to distinguish Ornithodoros brodyi and Ornithodoros yumatensis (Ixodida: Argasidae) in their different developmental stages.

Ornithodoros brodyi and Ornithodoros yumatensis are two species distributed in the Americas and associated with bats and caves. Both species have similar morphology, and the diagnostic traits of adults have not been detailed or illustrated accurately. In this study, the independence of both species is validated on the basis of molecular evidence (using partial sequences of 16S rDNA gene), and the morphological differences between them (dentition of the hypostome and traits of individual mammillae) are confirmed through light and scanning electron microscopy. In addition to the above characteristics, we observed other traits that may serve to differentiate both species: dorsal setae are short and thick in O. yumatensis and are thin and moderate in size in O. brodyi. We also observed a conspicuous hood in O. brodyi, which was absent in O. yumatensis. Another characteristic observed is a line of setae, near the end of Tarsi II-IV, which in O. brodyi is formed by less than five setae and in O. yumatensis by more than five. The main morphological difference between larvae of the 2 species is the number of ventral setae [9 (4 circumanal pairs) in O. brodyi and 8 (3 circumanal pairs) in O. yumatensis]. The genetic divergence in 16S rDNA sequences between these two species ranges from 9.7 to 10.6%.

Acaricidal activity of fluralaner against Ornithodoros moubata and Ornithodoros erraticus argasid ticks evaluated through in vitro feeding.

Ornithodoros erraticus and Ornithodoros moubata are argasid tick vectors that transmit severe diseases such as African swine fever and human relapsing fever. Elimination of the synanthropic populations of these vectors would facilitate the control of these diseases. Fluralaner is a novel isoxazoline that selectively blocks the GABA- and glutamate-gated channels, providing potent insecticidal and acaricidal activity. The aim of the current study was to provide quantitative data on the susceptibility of males, females and third nymphal instar of O. erraticus and O. moubata to fluralaner through in vitro feeding exposure. Fluralaner activity against these developmental stages and species was assessed by feeding the ticks on ovine blood medicated with decreasing fluralaner concentrations between 1 and 10-8µg/mL. Tick mortality was measured at 4, 24 and 48h and 1, 2 and 3 weeks post-feeding. Tests included solvent-treated and untreated blood controls. Fluralaner was extremely active against O. erraticus, with mean lethal concentrations 50 (LC50) and 95 (LC95) of 2.0×10-8 and 5.4×10-8µg/mL, respectively. Fluralaner was also highly active against O. moubata, showing a mean LC50 of 1.5×10-6µg/mL and a mean LC95 of 1.8×10-3µg/mL. In the latter species, the most susceptible life stages were the females (LC95 1.4×10-4µg/mL). Fluralaner demonstrated potent acaricidal activity against all developmental stages of O. erraticus and O. moubata tested, in the first 48h after in vitro feeding. Therefore, fluralaner has the potential to provide very high acaricidal efficacy to multiple argasid tick species via feeding exposure and could be included as an acaricidal agent in integrated programmes for the control of argasid tick vectors and argasid tick-borne diseases.

Redescription of Ornithodoros dyeri (Ixodida: Argasidae) based on morphologic and molecular data.

Larvae, nymphs and adults of the cave dwelling tick Ornithodoros dyeri, collected in 3 Mexican states, were studied using morphological and molecular methods. The adults and nymphs were characterized by an elongated body in proportion to the width and a dorsum bounded by two contiguous ridges and one third ridge (inner) that was incomplete on each side. The larvae of this species have 14 pairs of dorsal setae, a venter body with nine pairs of setae plus a posteromedian; a moderately large, dorsal plate and piriform, a hypostome arising from a relatively short, subtriangular median extension of the basis capituli, and a capsule of Hallers organ with reticulations. Based on a maximum likelihood analysis of the sequences of a fragment of approximately 414bp of the mitochondrial 16S rRNA gene, we showed that O. dyeri represents an independent lineage within neotropical species of the Argasidae. The bat species Mimon cozumelae and Peropteryx macrotis represents a new host record for this argasid.