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 Haller's 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.

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.

Description of nymphal instars of Ornithodoros mimon Kohls, Clifford & Jones, 1969 (Acari: Argasidae).

Ornithodoros mimon is an argasid tick common on Chiroptera in the Neotropical region, where it also bites humans aggressively. Here we describe for the first time all nymphal instars (N1, N2 and N3) of O. mimon based on optical and scanning electron microscopy. Although the nymphal instars of O. mimon resemble each other closely, there are characters that differentiate them: the N3 are taller that N1 and N2; the genital primordium occurs in some N2 and all N3; the spiracular plate in N1 and N2 is cone-like, but in N3 it is semicircular; and the submarginal dorsal groove is less distinct in N1 but more evident in N3. Nymphs of 0. mimon closely resemble the bat-associated species of the genus Ornithodoros included in the Alectorobius group. We review prior descriptions of nymphs of the Alectorobius group and make comparisons with nymphs of O. minon, highlighting characters with diagnostic information, such as the idiosomal shape, presence of discs and hood and absence subapical protuberance of tarsus I. The description of nymphal instars of O. mimon herein presented, improves the taxonomy of the family Argasidae, performing a work more detailed about the immature stage of this species.

Detection and identification of putative bacterial endosymbionts and endogenous viruses in tick cell lines.

As well as being vectors of many viral, bacterial, and protozoan pathogens of medical and veterinary importance, ticks harbour a variety of microorganisms which are not known to be pathogenic for vertebrate hosts. Continuous cell lines established from ixodid and argasid ticks could be infected with such endosymbiotic bacteria and endogenous viruses, but to date very few cell lines have been examined for their presence. DNA and RNA extracted from over 50 tick cell lines deposited in the Roslin Wellcome Trust Tick Cell Biobank (http://tickcells.roslin.ac.uk) were screened for presence of bacteria and RNA viruses, respectively. Sequencing of PCR products amplified using pan-16S rRNA primers revealed the presence of DNA sequences from bacterial endosymbionts in several cell lines derived from Amblyomma and Dermacentor spp. ticks. Identification to species level was attempted using Rickettsia- and Francisella-specific primers. Pan-Nairovirus primers amplified PCR products of uncertain specificity in cell lines derived from Rhipicephalus, Hyalomma, Ixodes, Carios, and Ornithodoros spp. ticks. Further characterisation attempted with primers specific for Crimean-Congo haemorrhagic fever virus segments confirmed the absence of this arbovirus in the cells. A set of pan-Flavivirus primers did not detect endogenous viruses in any of the cell lines. Transmission electron microscopy revealed the presence of endogenous reovirus-like viruses in many of the cell lines; only 4 of these lines gave positive results with primers specific for the tick Orbivirus St Croix River virus, indicating that there may be additional, as yet undescribed 'tick-only' viruses inhabiting tick cell lines.

Molecular identification and description of the female of Nothoaspis reddelli (Ixodida: Argasidae) from a cave in southeastern Mexico.

Nothoaspis reddelli Keirans and Clifford, 1975 , was described from 3 males collected in Grutas de Xtacumbilxunaán, Campeche, Mexico, although females have remained undescribed for 37 yr. Recently adult females of this species were collected from Cueva de Villa Luz ( =  Cueva de las Sardinas, Cueva del Azufre), in Tapijulapa, Tabasco, Mexico. Here we present a morphological description of the female stage, together with 16S rDNA sequences that confirm the conspecificity of our female, male, and nymphal specimens. The female integument of the anterior portion of the dorsal surface is smooth (nothoaspis), appearing to consist of 3 large "subunits," 1 anterior and 2 posterior, each with a small sublateral "subunit" on either side. The remaining dorsal covered integument is a cell-like configuration. The hood is large and bluntly rounded, and visible dorsally. The spiracular plate is oval. It possesses 1 pair of posthypostomal setae. The palpal trochanter has 1 pair of setae and a 5/5 hypostome decreasing to 4/4 at the apex. There is a single central pore at the base of the hypostome.

Developmental changes in salivary glands of nymphs and adults of the spinose ear tick Otobius megnini.

Otobius megnini has an autogenous 1 host life cycle, where larvae and nymphs stay attached inside the ear canal for long periods, but the adult tick is free living and can lay several egg batches without feeding. In order to obtain information about anatomical structures involved in this particular life cycle, nymphs and adults of O. megnini were dissected and salivary gland images were obtained in situ with the use of scanning electronic microscopy. Measurements of salivary alveoli were obtained with the use of ImageJ 1.40 g software. In the nymphs, the Type I alveoli are relatively small (mean diameter: 19.6 µm) compared with those of the adults (mean: 43.4 µm) and other soft ticks in the literature. Type II alveoli in nymphs are similar (mean: 82.6 µm) to previously described alveoli in adult soft ticks. In contrast, the adults of O. megnini Type II alveoli are smaller (mean: 36.8 µm) and have a wrinkled surface. These findings provide more evidence that Type I alveoli take part in absorption of moisture during the free-living tick stages.

Carios mimon (Acari: Argasidae): description of adults and redescription of larva.

Carios mimon is an argasid tick common on Chiroptera, originally described from larvae collected on bats Mimon crenulatum from Bolivia and Eptesicus brasiliensis from Uruguay. Later it was also registered from Argentina and recently included among the Brazilian tick fauna. In Brazil, this species is very aggressive to man, resulting in intense inflammatory response and pain. It is known only by the larval description and its morphology resembles that from other species currently included into the genus Carios, formerly classified into the subgenus Alectorobius, genus Ornithodoros. Here we describe adults and redescribe the larva of C. mimon, based on light and scanning electron microscopy. Remarks about its morphological similarity with other species of this genus are also discussed. Molecular analysis inferred from a portion of the 16S rRNA mitochondrial gene placed C. mimon in a cluster supported by maximal bootstrap value (100%) with other argasid species (mostly bat parasites in the New World), which have been classified into either the genus Ornithodoros or Carios, depending on the Argasidae classification adopted by different authors.

Description of a new argasid tick (Acari: Ixodida) from bat caves in Brazilian Amazon.

Nothoaspis amazoniensis n. sp. (Acari: Ixodida: Argasidae) is described from adult and immature ticks (nymph II, nymph I, larva) collected from bat caves in the Brazilian Amazon. Also, 16S rDNA sequences are provided. The diagnostic characters for adults are the presence of false shield or nothoaspis, an anteriorly projecting hood covering the capitulum, a medial extension of palpal article I (flaps), genital plate extending from coxa I to IV, absence of 2 setae on the internal margin of the flaps, a minute hypostome without denticles, presence of a central pore in the base of hypostome, and a reticulate surface pattern on the posterior half of the nothoaspis in males. The nymph II stage is characterized by a hood that is small in relation to the capitulum, short coxal setae, palpal flaps lacking setae on the internal margin, long hypostome, pointed with dentition 4/4 apically, and the anterior half of the body is covered by a cell-like configuration. Nymph I stage is characterized by a hood, small in relation to the capitulum, dorsum of the body covered by a cell-like configuration, venter integument covered by a cell-like configuration, and hypostome dentition 4/4 with apices that are "V"-shaped. Diagnostic characters of the larvae are the number and size of dorsal setae, and the shape of scutum and hypostome. The new species appears to have a life cycle with a larva that feeds on bats, a non-feeding nymphal stage (nymph I), a feeding nymphal stage (nymph II), and adults that probably represent non-feeding stages.

New reports of Antricola guglielmonei and Antricola delacruzi in Brazil, and a description of a new argasid species (Acari).

Adults of 3 tick species (Acari: Argasidae) identified as Antricola guglielmonei, Antricola delacruzi, and Carios rondoniensis n. sp. were collected on bat guano in a cave in the state of Rondônia, western Amazon, Brazil. Adults of C. rondoniensis possess a unique combination of characters that distinguish them from all described adults in the Argasidae, i.e., a large spiracular plate densely filled with small goblets, a well-developed flap covering the female genital opening, and palpi containing several tufts of long setae on articles 2 and 3. Unlike Ornithodoros or other Carios species, adults of C. rondoniensis have a scooplike hypostome devoid of denticles, as in Antricola spp. Conversely, the presence of a pair of long posthypostomal setae, and a slitlike transverse fissure at the capsule opening of the Haller's organ, are characters of C. rondonensis that are also found in species of Carios and Ornithodoros, but not in Antricola species. Molecular analyses inferred from a portion of the 16S rRNA mitochondrial gene indicate that C. rondoniensis is phylogenetically closest to species of Carios, followed by species of Antricola, and then Ornithodoros. Because the highest bootstrap value linking C. rondoniensis to Carios spp. was 62%, further phylogenetic studies are needed to better evaluate the taxonomic status of the former species.

Ultrastructure of normal and JHA-treated eggs of the soft tick Argas persicus (Oken).

The ultrastructure of normal development and JHA-treated egg was examined with the transmission electron microscope. In newly oviposited egg, the chorion consists of exo- and endochorion. Between the chorion and plasma membrane lies vitelline (envelope) membrane, a thin non cellular membrane. The periplasm is free of the yolk spheres with contains vesicles of various sizes and lipid inclusions. The endoplasm is rich in yolk spheres, mitochondria, lipid inclusions, dense vesicles and vacuoles. In 48 h-old egg, cleavage nuclei was seen migrating to the periphery of the egg. The ooplasm around the nuclei contains vesicles, endoplasmic reticulum and few mitochondria. The blastoderm cells contain many organelles and inclusions. Secondary vitellophages and pole cells were observed. In 96 h-old egg, the germ band is formed. It consists of two layers, ectoderm and mesoderm. Amnion and serosa were visible. At (0-1 hour) eggs post-treatment with JHA (Admiral), no real effects was observed except for vacuolation of endoplasm; while at 48 h & 96 h-old post-treatment of eggs, great deterioration were seen which ended by degeneration and lysis of cell components leaving cell debris, vacuolation of nuclei, iregular shape of nuclear membrane and dispersed chromatin material.

A reassessment of argasid tick salivary gland ultrastructure from an immuno-cytochemical perspective.

Previous morphological and histochemical studies of argasid tick salivary glands indicated that they were less complex than ixodid salivary glands, with only three granular cell types. The present study shows that there exist at least four different granular cell types in the salivary glands of the argasid tick Ornithodoros savignyi, based on immuno-localization of the anti-hemostatic factors, apyrase and savignygrin. Both anti-hemostatic factors were localized to dense core granule type 'a' and to granule type 'b', that shares a similar homogenous morphology with non-labeled granule type 'd'. Furthermore, the major tick salivary gland proteins (TSGPs), previously implicated in granule biogenesis, were localized to all the granular cell types. This indicates that granular cell types with different morphologies can express the same proteins, while cell types that show similar morphologies may not express the same proteins. Argasid tick salivary glands seem to be more complex than previously thought and might not be amenable to morphological classification alone. Alternative classification methodologies that rely on physical expression patterns of the salivary gland proteome might be more reliable as markers for a specific granular cell type.

Three new species of Antricola (Acari: Argasidae) from Brazil, with a key to the known species in the genus.

Three new species of Antricola (Acari: Argasidae) are described from adult specimens collected on bat guano in different caves in Brazil. The female of Antricola guglielmonei n. sp. is easily determined by the presence of 2 smooth, depressed areas lacking setae in the posterolateral portions of the dorsum of the idiosoma, together with the partial fusion of the tubercles in the anteromedian portion of the idiosoma. The male of this species has a small spiracular plate surrounded by a pattern of tubercles disposed concentrically in its dorsal portion. Both sexes have cervical grooves very slightly marked. Antricola delacruzi n. sp. represents the only Antricola species with the dorsum of the idiosoma devoid of tubercles in both sexes and with scarce and minute setae placed over the smooth cuticle. The female of A. inexpectata n. sp. is known only from a few specimens. In this species, lines of smooth cuticle lacking setae separate the tubercles of the dorsum. In addition, there are 3 clumps of plumose setae close to the spiracular plate, in the ventrolateral portions of the idiosoma, in 3 well-delimited regions over cuticular thickenings. These 3 species share the peculiarity of a Haller's organ with the anterior pit bearing only 7 + 2 setae. The collection of these new species in Brazilian caves greatly expands the known range of the genus. A key to the adults of all known species of the genus is provided.