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Background Amblyomma cajennense F. is one of the best known and studied ticks in the New World because of its very wide distribution, its economical importance as pest of domestic ungulates, and its association with a variety of animal and human pathogens. Recent observations, however, have challenged the taxonomic status of this tick and indicated that intraspecific cryptic speciation might be occurring. In the present study, we investigate the evolutionary and demographic history of this tick and examine its genetic structure based on the analyses of three mitochondrial (12SrDNA, d-loop, and COII) and one nuclear (ITS2) genes. Because A. cajennense is characterized by a typical trans-Amazonian distribution, lineage divergence dating is also performed to establish whether genetic diversity can be linked to dated vicariant events which shaped the topology of the Neotropics. Results Total evidence analyses of the concatenated mtDNA and nuclear + mtDNA datasets resulted in well-resolved and fully congruent reconstructions of the relationships within A. cajennense . The phylogenetic analyses consistently found A. cajennense to be monophyletic and to be separated into six genetic units defined by mutually exclusive haplotype compositions and habitat associations. Also, genetic divergence values showed that these lineages are as distinct from each other as recognized separate species of the same genus. The six clades are deeply split and node dating indicates that they started diverging in the middle-late Miocene. Conclusions Behavioral differences and the results of laboratory cross-breeding experiments had already indicated that A. cajennense might be a complex of distinct taxonomic units. The combined and congruent mitochondrial and nuclear genetic evidence from this study reveals that A. cajennense is an assembly of six distinct species which have evolved separately from each other since at least 13.2 million years ago (Mya) in the earliest and 3.3 Mya in the latest lineages. The temporal and spatial diversification modes of the six lineages overlap the phylogeographical history of other organisms with similar extant trans-Amazonian distributions and are consistent with the present prevailing hypothesis that Neotropical diversity often finds its origins in the Miocene, after the Andean uplift changed the topology and consequently the climate and ecology of the Neotropics.

The Chamela Biological Station (ChBS) is located in the Pacific Coast of Mexico in the State of Jalisco. This represents one of the core areas of the Chamela-Cuixmala Biosphere Reserve, characterized by one of the most threatened ecosystems, the tropical dry forest. Although accumulated knowledge of biological diversity, only few studies have been focused on ectoparasites or ectodytes associated with mammals, only 23 arthropod taxa had been recorded. In order to increase knowledge about arthropods associated with Mexican mammals, the objective of this work was to record the richness of arthropods (mites, ticks, lice, and fleas) associated with small and medium-sized mammals in the ChBS. A total of 81 hosts belonging to four orders, six families and nine species were captured. From these hosts, 4,946 arthropods were recovered: 4,007 mites, 673 ticks, 230 lice, and 36 fleas. Among medium-sized mammals, Nasua narica (L.) and Didelphis virginiana Kerr showed the highest levels of richness, with six arthropod taxa; among rodents, Heteromys pictus (Thomas) had the highest number of associated species (five). Within the 22 arthropod taxa registered in the present work, 12 represent new records for the reserve, and 3 represent new records for Mexico. With this study, the arthropod fauna associated with mammals in the ChBS has been raised to 38 taxa. In terms of biological conservation, knowledge of the species that inhabit natural reserves must be a priority, since this represents the baseline for species protected, not only in Mexico but around the world.

Based on tick specimens collected recently in Mexico, Nicaragua, Panama and Brazil, we provide morphological descriptions of the nymph and adults of Ornithodoros clarki Jones & Clifford, 1972 from the first three countries, and the larva and nymph of Ornithodoros rondoniensis (Labruna, Terassini, Camargo, Brandão, Ribeiro & Estrada-Peña, 2008) from Brazil. Also, an analysis of mitochondrial 16S rDNA sequences was performed to analyze the phylogenetic relationships of these tick species. Adults and nymphs of O. clarki and O. rondoniensis are unique among the Argasidae family by presenting exceptionally large spiracular plates with small goblets, and an integument with smooth polygonal mammillae. However, these two species are morphologically distinct based on specific patterns of coxal folds, idiosomal mammillae and pilosity, and female genital flap. In contrast, the larvae of O. clarki and O. rondoniensis are morphologically identical, except for a general larger size of the former species; this slight difference is corroborated by Principal Component Analysis (PCA) by using 40 morphometric variables. Phylogenetic analyses including 16S rDNA partial sequences of different Ornithodoros taxa from Central and South America indicate that O. rondoniensis from Brazil diverges from O. clarki from Mexico, Nicaragua and Panama. However, phylogenetic distance separating both alleged species is similar or slightly lower than the distances depicted for conspecific populations of a few other Ornithodoros species. Nonetheless, our primary criterion to maintain O. rondoniensis as a valid species is because its adult and nymphal stages do present distinct morphological traits that easily distinguish these postlarval stages from O. clarki.

Ehrlichia chaffeensis is a bacterium belonging to the Anaplasmataceae family. In Mexico, only 2 species have been recorded in association with tick species and humans. The objective of the present study was to detect the presence of bacteria of the genus Ehrlichia in ticks collected from the Chamela-Cuixmala Biosphere Reserve, Jalisco, Mexico. The collected ticks were identified and analyzed individually by polymerase chain reaction to amplify a fragment of the Anaplasmataceae 16S rRNA gene and the Ehrlichia-specific dsb gene. A total of 204 ticks, corresponding to 5 species of Ixodidae and 1 of Argasidae, were collected from 147 mammals of 6 species and 4 orders; 57 ticks collected from vegetation were also included. Among the total ticks collected, 1.47% (3/204) was positive for Ehrlichia sp. DNA was obtained using the primers EHR 16SD and EHR 16SR for 16S rRNA and DSB-330 and DSB-728 for dsb. The positive samples corresponded to a larva (Amblyomma sp.) associated with Didelphis virginiana and 2 nymphs (Amblyomma cf. oblongoguttatum) infesting Nasua narica. None of the ticks collected from the vegetation tested positive for Ehrlichia sp. DNA on the basis of the 16S rRNA and dsb genes. The sequences from the larvae of Amblyomma sp. and the nymphs of A. cf. oblongoguttatum were similar to those of E. chaffeensis. The phylogenetic analysis inferred with maximum likelihood corroborated the identity as E. chaffeensis. Although the role of these tick species as vectors of E. chaffeensis is still undetermined, the presence of infected ticks in the area indicates a potential zoonotic risk.

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.

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%.

Ehrlichia chaffeensis is a bacterium belonging to the Anaplasmataceae family. In Mexico, only 2 species have been recorded in association with tick species and humans. The objective of the present study was to detect the presence of bacteria of the genus Ehrlichia in ticks collected from the Chamela-Cuixmala Biosphere Reserve, Jalisco, Mexico. The collected ticks were identified and analyzed individually by polymerase chain reaction to amplify a fragment of the Anaplasmataceae 16S rRNA gene and the Ehrlichia-specific dsb gene. A total of 204 ticks, corresponding to 5 species of Ixodidae and 1 of Argasidae, were collected from 147 mammals of 6 species and 4 orders; 57 ticks collected from vegetation were also included. Among the total ticks collected, 1.47% (3/204) was positive for Ehrlichia sp. DNA was obtained using the primers EHR 16SD and EHR 16SR for 16S rRNA and DSB-330 and DSB-728 for dsb. The positive samples corresponded to a larva (Amblyomma sp.) associated with Didelphis virginiana and 2 nymphs (Amblyomma cf. oblongoguttatum) infesting Nasua narica. None of the ticks collected from the vegetation tested positive for Ehrlichia sp. DNA on the basis of the 16S rRNA and dsb genes. The sequences from the larvae of Amblyomma sp. and the nymphs of A. cf. oblongoguttatum were similar to those of E. chaffeensis. The phylogenetic analysis inferred with maximum likelihood corroborated the identity as E. chaffeensis. Although the role of these tick species as vectors of E. chaffeensis is still undetermined, the presence of infected ticks in the area indicates a potential zoonotic risk. Key words: bacteria, mammal, reserve, tick, disease

Ehrlichia chaffeensis is a bacterium belonging to the Anaplasmataceae family. In Mexico, only 2 species have been recorded in association with tick species and humans. The objective of the present study was to detect the presence of bacteria of the genus Ehrlichia in ticks collected from the Chamela-Cuixmala Biosphere Reserve, Jalisco, Mexico. The collected ticks were identified and analyzed individually by polymerase chain reaction to amplify a fragment of the Anaplasmataceae 16S rRNA gene and the Ehrlichia-specific dsb gene. A total of 204 ticks, corresponding to 5 species of Ixodidae and 1 of Argasidae, were collected from 147 mammals of 6 species and 4 orders; 57 ticks collected from vegetation were also included. Among the total ticks collected, 1.47% (3/204) was positive for Ehrlichia sp. DNA was obtained using the primers EHR 16SD and EHR 16SR for 16S rRNA and DSB-330 and DSB-728 for dsb. The positive samples corresponded to a larva (Amblyomma sp.) associated with Didelphis virginiana and 2 nymphs (Amblyomma cf. oblongoguttatum) infesting Nasua narica. None of the ticks collected from the vegetation tested positive for Ehrlichia sp. DNA on the basis of the 16S rRNA and dsb genes. The sequences from the larvae of Amblyomma sp. and the nymphs of A. cf. oblongoguttatum were similar to those of E. chaffeensis. The phylogenetic analysis inferred with maximum likelihood corroborated the identity as E. chaffeensis. Although the role of these tick species as vectors of E. chaffeensis is still undetermined, the presence of infected ticks in the area indicates a potential zoonotic risk.