Molecular survey of Rickettsia spp. in ticks infesting wild animals in six departments in Colombia.

Ticks are a globally distributed group of hematophagous ectoparasites that parasitize terrestrial vertebrates such as amphibians, reptiles, birds, and mammals. Ticks are vectors and reservoirs of pathogens that play an important role in wildlife and human health. Rickettsia is one of the bacteria transmitted by ticks, which some pathogenic species can cause rickettsiosis, a zoonotic disease that can cause serious harm to humans and animals. More information is necessary on the interactions between ticks and wildlife despite the fifty-seven ticks species already identified in Colombia. The objective of the present study was to determine the associations between ticks parasitizing wildlife and bacteria of the genus Rickettsia in six departments of Colombia. One hundred eighty-five ticks (80 larvae, 78 nymphs, and 27 adults) were collected from 55 wildlife species (amphibians, birds, mammals, and reptiles). Nine tick species were identified, and Rickettsia bellii, Rickettsia felis, 'Candidatus Rickettsia colombianensi' and Rickettsia parkeri were detected. Our results contribute to the current knowledge of tick-associated rickettsiae and the role of wildlife in their transmission dynamics.

Pathogens associated with ticks (Acari: Ixodidae) and mammals in the Orinoquia region of Colombia: An approach to understanding vector-pathogen-host interactions.

The hard tick clade (Ixodidae) currently comprises 762 species worldwide (266 Prostriata and 496 Metastriata). A quarter of hard ticks are found in the Neotropical region, and 42 species have been documented in Colombia. Ixodidae species are important vectors of pathogens such as bacteria, helminths, protozoa, and viruses. In tick-borne diseases, vertebrate hosts perform an important role in the transmission, maintenance, and spread of pathogens. Colombia ranks sixth among countries with the highest mammal biodiversity, with a total of 548 species, where some of these species may be involved in pathogen transmission cycles with ticks as vectors. This research evaluated the presence of two genera of bacteria (Borrelia and Rickettsia) and the protozoan (Babesia) in ticks and mammals in the Orinoquia region of Colombia, establishing interaction networks. The information comes from 734 mammals (655 wild and 79 domestic), belonging to 59 species. Tick infestation (n = 1,805) was found with 14.85 % (n = 109) of the examined mammals and corresponds to nine tick species confirmed morphologically and molecularly. To detect pathogens 272 ticks were collected while feeding on 96 mammals; samples from 93 mammals were analyzed. The presence of borreliae from the relapsing fever group (RFG) and the Lyme disease group (LDG) were detected. Rickettsia spp. was detected in ticks and mammals, while Babesia bigemina was only detected in ticks. This research is the first to address the prevalence of zoonotic pathogens in domestic and wild mammals infested with hard ticks in the Department of Arauca, Colombia. Considering that reporting cases of infections with Babesia, Borrelia, and Rickettsia in Colombia is not mandatory, their impact on public health cannot be estimated. This highlights the importance of continuously detecting, confirming, and identifying these and other important pathogens within the "One Health" framework, as they have a significant economic and medical-veterinary impact globally.

Environmental and ecological factors affecting tick infestation in wild birds of the Americas.

The Americas hold the greatest bird diversity worldwide. Likewise, ectoparasite diversity is remarkable, including ticks of the Argasidae and Ixodidae families - commonly associated with birds. Considering that ticks have potential health implications for humans, animals, and ecosystems, we conducted a systematic review to evaluate the effects of bioclimatic, geographic variables, and bird species richness on tick infestation on wild birds across the Americas. We identified 72 articles that met our inclusion criteria and provided data on tick prevalence in wild birds. Using Generalized Additive Models, we assessed the effect of environmental factors, such as habitat type, climatic conditions, bird species richness, and geographic location, on tick infestation. Our findings show that most bird infestation case studies involved immature ticks, such as larvae or nymphs, while adult ticks represented only 13% of case studies. We found birds infested by ticks of the genera Amblyomma (68%), Ixodes (22%), Haemaphysalis (5%), Dermacentor (1%), and Rhipicephalus (0.8%) in twelve countries across the Americas. Our findings revealed that temperature variation and bird species richness were negatively associated with tick infestation, which also varied with geographic location, increasing in mid-latitudes but declining in extreme latitudes. Our results highlight the importance of understanding how environmental and bird community factors influence tick infestation in wild birds across the Americas and the dynamics of tick-borne diseases and their impact on biodiversity.

Avian haemosporidians of the genera Plasmodium and Haemoproteus from resident and Neotropical migrant birds in Colombia.

Avian haemosporidians of the genera Plasmodium and Haemoproteus are a group of widely distributed blood parasites that can negatively affect the fitness of their hosts. Colombia contains the greatest diversity of birds on the planet, but knowledge about the associations between haemosporidian and its avifauna is scarce and fragmented. We collected blood samples from 255 birds (203 residents and 52 neotropical migrants) belonging to 27 families and 108 species. The study was conducted in six localities in the inter-Andean valleys of the Cauca and Magdalena rivers. Parasites of the genera Plasmodium and Haemoproteus were identified in the samples by morphological and molecular analysis of a fragment of the mitochondrial gene cyt b. Among the samples, 9.3% (n = 24) were positive for Plasmodium or Haemoproteus. Co-infection with Plasmodium and Haemoproteus was found in Red-eyed Vireo. Seventeen haemosporidian lineages were identified, five of which were reported for the first time in resident birds (Common Ground Dove, Checker-throated Stipplethroat, Tropical Kingbird, Pale-breasted Thrush, and Ruddy-breasted Seedeater) and one in the Summer Tanager (neotropical migrant). The research results confirm the wide diversity of haemosporidian present in tropical lowlands and the possible role of neotropical migratory birds in dissemination on haemosporidian along their migratory routes.

Bacterial community of ticks (Acari: Ixodidae) and mammals from Arauca, Colombian Orinoquia.

Ticks are obligate hematophagous ectoparasites of vertebrates and are relevant worldwide due to the number of bacterial and other pathogens they can transmit. To date, the knowledge about the microorganisms that ticks harbor and transmit to their hosts is incipient. In this study, 24 samples of mammals belonging to four taxonomic orders and ticks of the genera Amblyomma and Rhipicephalus from the Orinoco region of Colombia were analyzed to described and compare the bacterial microbiome. Genetic extraction was performed, and the V3-V4 region of the 16S rRNA gene was amplified by PCR. Libraries were created, and those samples with adequate quality indices were sequenced using Illumina MiSeq technology. Bacterial taxonomic assignment analyses were conducted through Amplicon Sequence Variants (ASVs) and Operational Taxonomic Units (OTUs). The results correspond to 16 samples that passed the quality filters, with 3218 OTUs (415 families). Although a considerable number of unknown bacteria was found, Enterobacteriaceae, Beijerinckiaceae, Moraxellaceae, and Burkholderiaceae are the most prevalent families, and the presence of the genera Coxiella, Escherichia-Shigella, Enterobacter, which can harbor pathogenic species was confirmed. In individuals of Amblyomma mixtum found actively feeding on Hydrochoerus hydrochaeris, bacteria of the genera Escherichia-Shigella and Enterobacter were documented. Similarly, Rhipicephalus microplus found actively feeding on Odocoileus virginianus cariacou shared Escherichia-Shigella. Ralstonia was shared among the blood samples of H. hydrochaeris, while Anaplasma and Eubacterium were shared in blood and liver samples of O. v. cariacou. Shared bacteria between A. mixtum and R. microplus included Bacillus, Coxiella, and Escherichia-Shigella. The results highlight the need of additional studies in other natural regions of Colombia and other American countries where tick-borne diseases have been detected. Likewise, the recorded data are the first at the level of bacterial communities in ticks of the family Ixodidae and provide valuable knowledge for the understanding host-tick and pathogen interactions.

Ticks (Acari: Ixodidae) on resident and migratory wild birds in Orinoquia region, Colombia.

Several species of hard ticks, including those of the genera Ixodes, Haemaphysalis, Amblyomma, and Rhipicephalus, are of medical and veterinary importance and have been reported in association with Neotropical wild birds. Colombia, known for its great bird diversity, has 57 confirmed tick species. However, there are few studies on the association between wild birds and ticks in Colombia. The Orinoquia region, a migratory center in Colombia, provides a unique opportunity to study wild bird-tick associations and their implications for tick-borne disease dynamics. Our study, conducted between October and December 2021, aimed to identify hard ticks infesting resident and migratory wild birds in the department of Arauca and to assess the presence of bacteria from the genera Anaplasma, Borrelia, Ehrlichia, Rickettsia, and piroplasms. A total of 383 birds were examined, of which 21 were infested. We collected 147 ticks, including Amblyomma dissimile (larvae), Amblyomma longirostre (nymphs), Amblyomma mixtum (adults), and Amblyomma nodosum (larvae and nymphs). We did not detect bacterial DNA in the tested ticks; however, piroplasm DNA was detected in ticks from three of the infested birds. Of the 21 bird-tick associations, six are new to the Americas, and interesting documentation of piroplasm DNA in A. longirostre, A. nodosum, and A. dissimile ticks from wild birds in the region. This study provides valuable insights into the ticks associated with wild birds and their role in the dispersal of ticks and pathogens in Colombia, enhancing our understanding of tick life cycles and tick-borne disease dynamics.

A new species of terrestrial toad of the Rhinellafestae group (Anura, Bufonidae) from the highlands of the Central Cordillera of the Andes of Colombia.

The genus Rhinella (Bufonidae) comprises 92 species of Neotropical toads. In Colombia, Rhinella is represented by 22 recognized species, of which nine belong to the Rhinellafestae group. Over the past decade, there has been increasing evidence of cryptic diversity within this group, particularly in the context of Andean forms. Specimens of Rhinella collected in high Andean forests on both slopes of the Central Cordillera in Colombia belong to an undescribed species, Rhinellakumandaysp. nov. Genetic analyses using the mitochondrial 16S rRNA gene indicated that the individuals belong to the festae species group. However, they can be distinguished from other closely related species such as Rhinellaparaguas and Rhinellatenrec by a combination of morphological traits including the presence of tarsal fold, a moderate body size, and substantial genetic divergence in the 16S rRNA gene (> 5%). Through this integrative approach, the specimens from the Central Cordillera of Colombia are considered an evolutionary divergent lineage that is sister to R.paraguas, and described as a new species. Rhinellakumandaysp. nov. is restricted to the Central Cordillera of Colombia inhabiting both slopes in the departments of Caldas and Tolima, in an elevational range between 2420 and 3758 m. With the recognition of this new species, the genus Rhinella now comprises 93 species with 23 of them found in Colombia, and ten species endemic to the country.

New insights into the molecular phylogeny, biogeographical history, and diversification of Amblyomma ticks (Acari: Ixodidae) based on mitogenomes and nuclear sequences.

BACKGROUND: Amblyomma is the third most diversified genus of Ixodidae that is distributed across the Indomalayan, Afrotropical, Australasian (IAA), Nearctic and Neotropical biogeographic ecoregions, reaching in the Neotropic its highest diversity. There have been hints in previously published phylogenetic trees from mitochondrial genome, nuclear rRNA, from combinations of both and morphology that the Australasian Amblyomma or the Australasian Amblyomma plus the Amblyomma species from the southern cone of South America, might be sister-group to the Amblyomma of the rest of the world. However, a stable phylogenetic framework of Amblyomma for a better understanding of the biogeographic patterns underpinning its diversification is lacking. METHODS: We used genomic techniques to sequence complete and nearly complete mitochondrial genomes -ca. 15 kbp- as well as the nuclear ribosomal cluster -ca. 8 kbp- for 17 Amblyomma ticks in order to study the phylogeny and biogeographic pattern of the genus Amblyomma, with particular emphasis on the Neotropical region. The new genomic information generated here together with genomic information available on 43 ticks (22 other Amblyomma species and 21 other hard ticks-as outgroup-) were used to perform probabilistic methods of phylogenetic and biogeographic inferences and time-tree estimation using biogeographic dates. RESULTS: In the present paper, we present the strongest evidence yet that Australasian Amblyomma may indeed be the sister-group to the Amblyomma of the rest of the world (species that occur mainly in the Neotropical and Afrotropical zoogeographic regions). Our results showed that all Amblyomma subgenera (Cernyomma, Anastosiella, Xiphiastor, Adenopleura, Aponomma and Dermiomma) are not monophyletic, except for Walkeriana and Amblyomma. Likewise, our best biogeographic scenario supports the origin of Amblyomma and its posterior diversification in the southern hemisphere at 47.8 and 36.8 Mya, respectively. This diversification could be associated with the end of the connection of Australasia and Neotropical ecoregions by the Antarctic land bridge. Also, the biogeographic analyses let us see the colonization patterns of some neotropical Amblyomma species to the Nearctic. CONCLUSIONS: We found strong evidence that the main theater of diversification of Amblyomma was the southern hemisphere, potentially driven by the Antarctic Bridge's intermittent connection in the late Eocene. In addition, the subgeneric classification of Amblyomma lacks evolutionary support. Future studies using denser taxonomic sampling may lead to new findings on the phylogenetic relationships and biogeographic history of Amblyomma genus.

Exploring the relationship between bats (Mammalia, Chiroptera) and ectoparasitic flies (Diptera, Hippoboscoidea) of the Orinoquia Region in South America.

Bat flies (Nycteribiidae and Streblidae) have been used to study co-evolutionary patterns between ectoparasites and bats. In the world, Nycteribiidae and Streblidae are represented by approximately 276 and 237 species, respectively. In regions such as the Orinoquia located in the north of South America (Colombia and Venezuela), the richness of bats is high (more than 100 documented species), but studies on Nycteribiidae and Streblidae are scarce and discontinuous. To contribute to the knowledge of ectoparasitic flies in the Orinoquia, records of flies and their interactions with bats were reviewed, including new records and associations using interaction networks. We documented 124 species of Streblidae and only 12 of Nycteribiidae for the Orinoquia in approximately 102 bat species reported in Colombia and Venezuela. New records for six species of bat flies in Colombia were found (Mastopteraguimaraesi, Noctiliostreblamaai, Paradyschiriaparvuloides, Trichobiusjubatus, Trichobiusparasiticus, and Basiliaferrisi) associated with six species of bats (Cynomopsplanirostris, Desmodusrotundus, Myotishandleyi, Molossusrufus, Noctilioalbiventris, and Phyllostomushastatus). The bat-ectoparasite interaction networks in the Orinoquia revealed a pattern of antagonistic relationships, with high specialization, modularity, and low connectivity and nesting. The identified networks are between bat fly species belonging to different ecomorphological groups with unique host species. This supports the idea of ecological niche partitioning among ectoparasitic bat flies and hosts. Our study expanded the knowledge of the distribution of some fly species and the associations with bat hosts in Colombia, by presenting morphological descriptions and new observations, which are key to understanding the ecology, diversity, and distribution of these species.

Phylogenetic relationships of the Amblyomma cajennense complex (Acari: Ixodidae) at mitogenomic resolution.

The genus Amblyomma is the third most diverse in the number of species within the Ixodidae, with practically half of its species distributed in the Americas, though there are also species occurring in Africa, Asia, and Australia. Within the genus, there are several species complexes with veterinary and public health importance. The Amblyomma cajennense complex, in the Americas, is represented by six species with a wide distribution, from Texas to northern Argentina. We combined two sequencing techniques to generate complete mitogenomes of species belonging to the Amblyomma cajennense complex: genome skimming and long-range PCRs sequencing methods. Thus, we generated seven new mitochondrial genomes for all species of the Amblyomma cajennense complex, except for Amblyomma interandinum. Genetic distances between the mitogenomes corroborate the clear differentiation between the five species of the Amblyomma cajennense complex. The phylogenetic relationships of these species had previously been evaluated by combining partial nuclear and mitochondrial genes and here these relationships are corroborated with a more robust framework of data, which demonstrates that the conjunction of mitochondrial and nuclear partial genes can resolve close relationships when entire genes or genomes are unavailable. The gene order, structure, composition, and length are stable across these mitogenomes, and they share the general characteristics of Metastriata. Future studies should increase the number of available mitogenomes for this genus, especially for those species from the Indo-Pacific region and Africa, by means of a better understanding of their relationships and evolutionary process.

Morphological and molecular confirmation of Ornithodoros hasei (Schulze, 1935) (Acari: Argasidae) in Colombia.

A large number of tick species are proven vectors for the transmission of bacteria, protozoa, and viruses. Soft ticks (Acari: Argasidae) in South America have been found to be the most frequent carriers of borreliae of the relapsing fever group (RFG); however, there are several information gaps specially on the taxonomy and distribution of some tick species. Here, we used light microscopy, scanning electron microscopy, and PCR amplification of a fragment of the mitochondrial 16S rRNA gene to evaluate 174 larvae of Ornithodoros (Argasidae) collected from three bat species (Eptesicus orinocensis, Molossus rufus and Noctilio albiventris) in the Orinoquia Region of Colombia. The morphological and molecular results confirmed that all the analyzed larvae corresponded to Ornithodoros hasei. Comparisons of mitochondrial 16S rDNA sequences showed low genetic divergence (0% - 0.3%) between larvae of the Department of Arauca in the Orinoquia Region and higher genetic divergence (3.4 - 4.7%) in sequences from other American countries. Our work represents the most recent collection of this species in Colombia and provides a molecular evaluation for the first time. Moreover, a new association of O. hasei with bats such as E. orinocensis is documented. Considering the wide distribution of O. hasei in the American Continent, and its putative role as vector for Borrelia, integrative studies that involve morphological, morphometric, molecular data and experimental crosses are needed to determine if the higher genetic distances are associated with cryptic speciation, as detected in other tick complexes, or represent genetic divergences among geographically different populations of O. hasei.

Confirming the presence of Lasiurusfrantzii (Peters, 1870) (Chiroptera, Vespertilionidae) in South America: more questions than answers.

The western or desert red bat, Lasiurusfrantzii, is a cryptic insectivore species distributed in the Neotropics from Mexico south through Central America to Panama. L.frantzii was long considered a subspecies of the red bat, Lasiurusblossevillii, but recently it was elevated to full-species status based on genetic information. Here we present evidence of the presence of L.frantzii in the Andean Region of Colombia, confirming the species' presence in South America; the new record, from 3836 m a.s.l., is also the highest elevation known for the species. We suggest that L.frantzii might be widely distributed in trans-Andean areas of Colombia, Ecuador, Venezuela, and perhaps Peru and Bolivia. However, a review and exploration of additional morphological traits to identify the species are necessary because of the uncertainty of the distribution of L.frantzii.

Revising the diversity within the Dwarf Dog-faced Bat, Molossopstemminckii (Chiroptera, Molossidae), with the revalidation of the endangered Molossopsgriseiventer.

The genus Molossops includes two monotypic species of insectivore bats distributed in South America: Molossopsneglectus and Molossopstemminckii. Both can be differentiated, based on sizes, M.temminckii being smaller (forearm less than 33 mm). Despite being monotypic, at least two additional subspecies have been described for M.temminckii, of which M.temminckiigriseiventer from the inter-Andean Valley of the Magdalena River in Colombia might represent a valid taxon. To test the taxonomic status of M.t.griseiventer, we reviewed specimens of M.temminckii from cis- and trans-Andean localities in Colombia. We used Cytochrome-b and Cytochrome Oxidase I comparisons to test the phylogenetic position of cis- and trans-Andean samples and compared qualitative morphology, morphometric and bioacoustics. Our results show that M.t.griseiventer is differentiated from cis-Andean specimens, providing further evidence of its validity at the species level. Furthermore, M.temminckii (sensu stricto) is also distributed in Colombia, but both M.griseiventer and M.temminckii are allopatric, with the Andes acting as a barrier. The specific identity of the specimens from the Caribbean Region of Colombia needs a new evaluation, but our results clearly show that the diversity of Molossops is underestimated.

Is Borrelia burgdorferi Sensu Stricto in South America? First Molecular Evidence of Its Presence in Colombia.

The genus Borrelia encompasses spirochetal species that are part of three well-defined groups. Two of these groups contain pathogens that affect humans: the group causing Lyme disease (LDG) and the relapsing fever group (RFG). Lyme disease is caused by Borrelia burgdorferi s.l., which is distributed in the Northern Hemisphere, and relapsing fevers are caused by Borrelia spp., which are found in temperate and tropical countries and are an emerging but neglected pathogens. In some departments of Colombia, there are records of the presence of Borrelia sp. in humans and bats. However, little is known about the impact and circulation of Borrelia spp. in the country, especially in wildlife, which can act as a reservoir and/or amplifying host. In this context, the objective of our research was to detect and identify the Borrelia species present in wild mammals in the departments of Caldas and Risaralda in Colombia. For morphological detection, blood smears and organ imprints were performed, and molecular identification was carried out through a nested PCR directed on the flagellin B (flaB) gene. A total of 105 mammals belonging to three orders (Chiroptera, Didelphimorphia and Rodentia) were analyzed, of which 15.24% (n = 16) were positive for Borrelia. Molecularly, the presence of Borrelia burgdorferi s.s. in lung tissues of Thomasomys aureus and blood of Mus musculus (Rodentia) was detected, with 99.64 and 100% identity, respectively. Borrelia sp. genospecies from a clade branch of a bat-associated LDG sister group were identified in seven individuals of bat species, such as Artibeus lituratus, Carollia brevicauda, Sturnira erythromos, and Glossophaga soricina. Furthermore, two Borrelia genospecies from the RFG in seven individuals of bats (A. lituratus, Artibeus jamaicensis, Platyrrhinus helleri, Mesophylla macconnelli, Rhynchonycteris naso) and rodents (Coendou rufescens, Microryzomys altissimus) were documented. Additionally, the presence of a spirochete was detected by microscopy in the liver of a Sturnira erythromos bat specimen. These results contain the first molecular evidence of the presence of B. burgdorferi s.s. in South America, which merits the need for comprehensive studies involving arthropods and vertebrates (including humans) in other departments of Colombia, as well as neighboring countries, to understand the current status of the circulation of Borrelia spp. in South America.

Infestation, histology, and molecular confirmation of Sarcoptes scabiei in an Andean porcupine (Coendou quichua) from the Central Andes of Colombia.

Sarcoptic mange is a highly contagious, worldwide disease that affects the skin of mammals, including humans. It is caused by the mite Sarcoptes scabiei, however, the information available in wild mammal populations in the world, and particularly in Colombia is limited. Here, we document a new case of sarcoptic mange in an Andean porcupine (Coendou quichua) from the Andean region of Colombia. We morphologically and molecularly confirmed the mite as S. scabiei and documented the histopathology associated with scabies, and show the different stages of the life cycle of S. scabiei from the Andean porcupine skin samples. Our review of reports of additional cases of scabies in wild mammal species in South America showed 15 species, mostly carnivores, artiodactyls, and rodents. Considering the limited information in Colombia, it is urgent to evaluate the risk of this condition on mammals which would contribute to the epidemiological knowledge and the potential implications of sarcoptic mange in the ecology and conservation of wild mammals in the country.

Bacteria related to tick-borne pathogen assemblages in Ornithodoros cf. hasei (Acari: Argasidae) and blood of the wild mammal hosts in the Orinoquia region, Colombia.

Interest in research on soft ticks has increased in recent decades, leading to valuable insight into their role as disease vectors. The use of metagenomics-based analyses have helped to elucidate ecological factors involved in pathogen, vector, and host dynamics. To understand the main bacterial assemblages present in Ornithodoros cf. hasei and its mammalian hosts, 84 ticks and 13 blood samples from bat hosts (Chiroptera) were selected, and the 16S rRNA gene V4 region was sequenced in five pools (each one related to each host-tick pairing). Bacterial taxonomic assignment analyses were performed by comparing operational taxonomic units (OTUs) shared between ticks and their host blood. This analysis showed the presence of Proteobacteria (38.8%), Enterobacteriaceae (25%), Firmicutes (12.3%), and Actinobacteria (10.9%) within blood samples, and Rickettsiaceae (39%), Firmicutes (25%), Actinobacteria (13.1%), and Proteobacteria (9%) within ticks. Species related to potentially pathogenic genera were detected in ticks, such as Borrelia sp., Bartonella tamiae, Ehrlichia sp. and Rickettsia-like endosymbiont, and the presence of these organisms was found in all analyzed bat species (Cynomops planirostris, Molossus pretiosus, Noctilio albiventris), and O. cf. hasei. About 41-48.6% of bacterial OTUs (genera and species) were shared between ticks and the blood of bat hosts. Targeted metagenomic screening techniques allowed the detection of tick-associated pathogens for O. cf. hasei and small mammals for the first time, enabling future research on many of these pathogens.

Prevalence of rickettsias in ticks (Acari: Ixodidae) collected in domestic animals of the northern region of Caldas department, Colombia / Prevalencia de rickettsias en garrapatas (Acari: Ixodidae) colectadas en animales domésticos del norte del departamento de Caldas, Colombia

ABSTRACT The northern region of the Caldas department in Colombia is considered an endemic area for murine typhus. Recent studies in patients with acute febrile disease demonstrated infection with the spotted fever group's rickettsiae due to an increase in the IgG titer by indirect immunofluorescence in paired sera obtained from these patients. The objective of the current research was to identify the species of ticks present in domestic animals in the northern region of Caldas and establish the presence of rickettsial genomic material in the collected ticks. Ticks were obtained from bovines, horses, and dogs in seven municipalities in the north of Caldas. Ticks were stored in 90 % ethanol until processing and were identified using taxonomic keys, DNA was extracted using commercial techniques, and the gltA gene was amplified by conventional chain reaction polymerase (PCR). Seven hundred thirteen ticks were obtained from 593 domestic animals. The highest infestation occurred in cattle, followed by canines and horses. Ticks found corresponded to the species Riphicephalus microplus, Dermacentor nitens, Amblyomma sp., and Riphicephalus sanguineus s.l. In none of the tick samples, Rickettsia-specific gltA gene DNA was found. It can be inferred that the ticks obtained are not a source of rickettsial infection for people in this department region, despite finding different species associated with the transmission of this disease.

RESUMEN La región norte del departamento de Caldas, Colombia es considerada como una zona endémica de tifo murino. Estudios recientes realizados en pacientes con enfermedad febril aguda, demostraron la infección con rickettsias, del grupo de las fiebres manchadas, debido al aumento en el título de IgG, por inmunofluorescencia indirecta (IFI), en sueros pareados, obtenidos de dichos pacientes. El objetivo de la investigación fue el de identificar las especies de garrapatas presentes en animales domésticos, de la región norte de Caldas y establecer la presencia de material genómico de rickettsias, en las garrapatas recolectadas. En siete municipios, se recolectaron garrapatas de bovinos, de equinos y de caninos. Las garrapatas, se almacenaron en etanol al 90 %, hasta su identificación taxonómica. Se extrajo el ADN, utilizando técnicas comerciales y se amplificó por reacción de cadena de la polimerasa (PCR) convencional el gen gltA. Se obtuvieron 713 garrapatas de 593 animales domésticos. La más alta infestación se presentó en bovinos, seguido de los caninos y equinos. Las garrapatas encontradas correspondieron a las especies Riphicephalus microplus, Dermacentor nitens, Amblyomma sp. y Riphicephalus sanguineus s.l. En ninguna de las muestras, se comprobó la presencia de ADN del gen gltA específico de Rickettsia. Se puede inferir que las garrapatas obtenidas no serían una fuente de infección rickettsial para las personas, en esta región del departamento; sin embargo, su presencia es un factor de riesgo para la adquisición de rickettsiosis asociadas con las fiebres manchadas.

Seroprevalence and detection of Rickettsia spp. in wild birds of Arauca, Orinoquia region, Colombia.

Wild birds have an important role as hosts of ticks infected by rickettsiae. However, the role of birds as reservoirs of tick-borne rickettsiae is unknown and poorly understood. This is particularly relevant in several tropical and subtropical areas, where migration influences the global spread of ectoparasites and pathogens of public health importance. This research aimed to detect and evaluate the exposure to spotted fever group rickettsiae in wild birds that could represent reservoirs in the Department of Arauca in the Colombian Orinoquia region. Sampling was conducted in three municipalities of the Department of Arauca (Colombia). Blood samples were collected from 255 birds and processed to obtain serum (n = 155) and DNA (n = 255) samples. The serum samples were processed for indirect immunofluorescence assays (IFA) for the detection of antibodies to Rickettsia rickettsii, Rickettsia parkeri, Rickettsia amblyommatis, Rickettsia rhipicephali, and Rickettsia bellii. Additionally, we investigated rickettsiae DNA in blood samples by amplification of the citrate synthase gene (gltA). The IFA results revealed seropositivity in 97 samples from 54 species of resident and migratory birds. No sample was positive for rickettsial DNA. The presence of antibodies in 62.5% of the sera indicates previous exposure of these birds to rickettsiae. The null detection of rickettsiae in the blood of seropositive birds is possibly due to a short period of bacteremia. Experimental studies are required to improve our understanding of the role of wild birds as sources of rickettsial infections in ticks.

Avian haemosporidian (Haemosporida: Plasmodium and Haemoproteus) in the department of Arauca, Colombian Orinoquia region.

Avian haemosporidian from the genera Haemoproteus and Plasmodium are a diverse and widely distributed group of vector-borne blood parasites. These parasites can have negative effects on bird survival by influencing several aspects of their life cycle, causing different clinical signs and even death. Colombia has the widest range of bird richness throughout the globe; however, the associations between haemosporidian parasites and wild birds in different ecosystems remain poorly explored. Within this frame of reference, the objective of the present study was to identify and understand haemosporidian associated with resident and migratory wild birds and their lineages in northeast Orinoquia region, Colombia. Birds were captured in 8 localities and blood samples were collected, identifying the presence of haemosporidian parasites through morphological and molecular analysis of the mitochondrial cytochrome b gene. Blood samples from 233 wild birds (86 species) were analyzed for haemosporidian parasites. Sixteen individuals (6.9%) from 15 resident and migratory species were positive for Haemoproteus or Plasmodium. Fourteen haemosporidian lineages were identified, five of them reported for the first time. These new lineages are reported in four resident birds and one boreal migratory bird (Parkesia noveboracensis). This study is the first developed in the department of Arauca and contributes to the knowledge of haemosporidian lineages and their interaction with wild birds in the Colombian Orinoquia region and South America.

Extension of the distribution of Amblyomma triste Koch, 1844: Morphological and molecular confirmation of Morphotype I in Colombia.

Ticks of the Amblyomma maculatum (Acari: Ixodidae) group are represented by three species: Amblyomma maculatum, Amblyomma tigrinum, and Amblyomma triste. This group is widely distributed throughout the Nearctic and Neotropical regions, from the United States of America (USA) to Argentina. They parasitize several vertebrates, including domestic mammals and humans. Nonetheless, the identification and distribution of the species within the group have been controversial but can be solved with integrative studies on their taxonomy and ecology. In Colombia, a Neotropical country from which up to 58 tick species have been reported, only Amblyomma maculatum Koch, 1844 (sensu stricto - s.s.) - morphotype II - has been molecularly confirmed. Here, specimens of the A. maculatum group from three departments of Colombia were examined by morphological and molecular analyses. The results confirmed the wide distribution of A. maculatum s.s. in Colombia. Furthermore, the presence of A. triste s.s. - morphotype I was confirmed for the first time at the molecular level, which broadens the distribution of this species in the American continent. Considering that both taxa are vectors of pathogens such as Rickettsia parkeri s.s., additional taxonomic, reproductive compatibility and niche distribution studies are needed, as well as the application of new molecular markers due to the controversial taxonomy. This will allow us to explore cryptic diversity within the recently established putative groups.