Isolation and molecular characterization of a novel relapsing fever group Borrelia from the white-eared opossum Didelphis albiventris in Brazil.

This study aimed to detect, isolate and to characterize by molecular methods a relapsing fever group (RFG) Borrelia in white-eared opossums (Didelphis albiventris) from Brazil. During 2015-2018, when opossums (Didelphis spp.) were captured in six municipalities of the state of São Paulo, Brazil, molecular analyses revealed the presence of a novel RFG Borrelia sp. in the blood of seven opossums (Didelphis albiventris), out of 142 sampled opossums (4.9% infection rate). All seven infected opossums were from a single location (Ribeirão Preto municipality). In a subsequent field study in Ribeirão Preto during 2021, two new opossums (D. albiventris) were captured, of which one contained borrelial DNA in its blood. Macerated tissues from this infected opossum were inoculated into laboratory animals (rodents and rabbits) and two big-eared opossums (Didelphis aurita), which had blood samples examined daily via dark-field microscopy. No spirochetes were visualized in the blood of the laboratory animals. Contrastingly, spirochetes were visualized in the blood of the two D. aurita opossums between 12 and 25 days after inoculation. Blood samples from these opossums were used for a multi-locus sequencing typing (MLST) based on six borrelial loci. Phylogenies inferred from MLST genes positioned the sequenced Borrelia genotype into the RFG borreliae clade basally to borreliae of the Asian-African group, forming a monophyletic group with another Brazilian isolate, "Candidatus B. caatinga". Based on this concatenated phylogenetic analysis, which supports that the new borrelial isolate corresponds to a putative new species, we propose the name "Candidatus Borrelia mimona".

Molecular survey of tick-borne infectious agents in cricetid rodents (Rodentia: Cricetidae) in Central and Southern Chile.

Tick-borne infectious agents (TBIAs) include several bacteria and protozoa that can infect vertebrates, including humans. Some of these agents can cause important diseases from both a public health perspective, such as Lyme disease, and from an animal health and production viewpoint, such as Texas fever. In Chile, several studies have assessed the presence of tick-borne disease agents in vectors and mammal hosts, mainly in the northern regions, but few studies have assessed the presence of these agents in Central and Southern Chile. This study aimed to assess the presence of three groups of TBIAs-Borrelia, Anaplasmataceae, and Piroplasmida-in cricetid rodents of Central and Southern Chile. A total of 207 specimens from 13 localities between the O'Higgins and Los Lagos regions were captured. DNA was extracted from the liver and spleen, and subsequently underwent polymerase chain reaction (PCR) amplification targeting the 16S rRNA, flaB, and 18S rRNA genes to detect DNA from Borrelia, Anaplasmataceae, and Piroplasmida, respectively. Although no DNA from these TBIAs was detected, the DNA extraction process was validated by optimal DNA purity ratios (an A260/A280 ratio within the 1.6-2.0 range) and successful internal control amplification in all samples. These results, in addition to findings from previous reports, suggest a very low prevalence of these TBIAs in the rodent population studied. Further research into the factors influencing the presence of these agents and their vectors will provide insight into the reasons underlying this low prevalence.

Hepatozoon (Eucoccidiorida: Hepatozoidae) in wild mammals of the Americas: a systematic review.

BACKGROUND: The study of parasites provides insight into intricate ecological relationships in ecosystem dynamics, food web structures, and evolution on multiple scales. Hepatozoon Eucoccidiorida: Hepatozoidae) is a genus of protozoan hemoparasites with heteroxenous life cycles that switch infections between vertebrates and blood-feeding invertebrates. The most comprehensive review of the genus was published 26 years ago, and currently there are no harmonized data on the epizootiology, diagnostics, genotyping methods, evolutionary relationships, and genetic diversity of Hepatozoon in the Americas. METHODS: Here, we provide a comprehensive review based on the PRISMA method regarding Hepatozoon in wild mammals within the American continent, in order to generate a framework for future research. RESULTS: 11 out of the 35 countries of the Americas (31.4%) had data on Hepatozoon, with Carnivora and Rodentia orders having the most characterizations. Bats, ungulates, and shrews were the least affected groups. While Hepatozoon americanum, H. americanum-like, H. canis, H. didelphydis, H. felis, H. milleri, H. griseisciuri, and H. procyonis correspond to the identified species, a plethora of genospecies is pending for a formal description combining morphology and genetics. Most of the vectors of Hepatozoon in the Americas are unknown, but some flea, mite, and tick species have been confirmed. The detection of Hepatozoon has relied mostly on conventional polymerase chain reaction (PCR), and the implementation of specific real time PCR for the genus needs to be employed to improve its diagnosis in wild animals in the future. From a genetic perspective, the V4 region of the 18S rRNA gene has been widely sequenced for the identification of Hepatozoon in wild animals. However, mitochondrial and apicoplast markers should also be targeted to truly determine different species in the genus. A phylogenetic analysis of herein retrieved 18S ribosomal DNA (rDNA) sequences showed two main clades of Hepatozoon: Clade I associated with small mammals, birds, and herpetozoa, and Clade II associated with Carnivora. The topology of the tree is also reflected in the haplotype network. CONCLUSIONS: Finally, our review emphasizes Hepatozoon as a potential disease agent in threatened wild mammals and the role of wild canids as spreaders of Hepatozoon infections in the Americas.

Experimental transmission of a novel relapsing fever group Borrelia harbored by Ornithodoros octodontus (Ixodida: Argasidae) in Chile.

Tick-borne relapsing fever spirochetes of genus Borrelia thrive in enzootic cycles involving Ornithodoros spp. (Argasidae) mainly, and rodents. The isolation of these spirochetes usually involves a murine model in which ticks are fed and the spirochetes detected in blood several days later. Such an experiment also demonstrates that a given species of tick is competent in the transmission of the bacteria. Here, soft ticks Ornithodoros octodontus were collected in Northern Chile with the objective to experimentally determine its capacity to transmit a Borrelia sp. detected in a previous study. Two Guinea pigs (Cavia porcellus) were used to feed nymphs and adults of O. octodontus and the spirochetes in blood were inspected by dark-field microscopy and nested PCR. Although spirochetes were not seen in blood, DNA was detected in only one animal 11 days after the ticks were fed. Genetic sequences of Borrelia flaB, clpX, pepX, recG, rplB, and uvrA genes retrieved from DNA extraction of positive blood were employed to construct two phylogenetic analyses. On the one hand, the flaB tree showed the Borrelia sp. transmitted by O. octodontus clustering with Borrelia sp. Alcohuaz, which was previously detected in that same tick species. On the other hand, concatenated clpX-pepX-recG-rplB-uvrA demonstrated that the characterized spirochete branches together with "Candidatus Borrelia caatinga", a recently discovered species from Brazil. Based on the genetic profile presented in this study, the name "Candidatus Borrelia octodonta" is proposed for the species transmitted by O. octodontus. The fact that spirochetes were not observed in blood of guinea pigs, may reflect the occurrence of low spirochetemia, which could be explained because the susceptibility of infection varies depending on the rodent species that is used in experimental models. Although the vertebrate reservoir of "Ca. Borrelia octodonta" is still unknown, Octodon degus, a rodent species that is commonly parasitized by O. octodontus, should be a future target to elucidate this issue.

A novel Babesia sp. of the "Western Babesia group", detected in opossums from Guatemala.

Babesia spp. are tick-borne protozoans that involve birds and mammals in their transmission cycles and cause babesiosis, a severe hemolytic malaria-like disease. Opossums of the genus Didelphis are recognized hosts of tick-borne pathogens. Therefore, exploring tick-borne agents in Didelphis species is important to understand the circulation of pathogens in areas where opossums occur. In this study, we targeted Anaplasmataceae, Babesia, Borrelia and Hepatozoon DNA in ticks, blood and organ samples collected from three hunted Didelphis marsupialis specimens in eastern Guatemala. While the samples were negative for Hepatozoon and bacterial DNA, sequences of Babesia 18S rDNA, cox1 and cytb genes were retrieved from two opossums. Ticks collected on the animals included Amblyomma parvum and an undetermined Ornithodoros sp. The Babesia sp. detected in this study (Babesia sp. THB1-2) clusters phylogenetically within the "Western Babesia group", which includes pathogenic species such as Babesia conradae, Babesia duncani, and Babesia negevi. Our results represent the first record of a Babesia sp. in Guatemala and highlight the importance of D. marsupialis as potential spreaders of ticks and pathogens in Central America.

Survey and Molecular Characterization of Sarcocystidae protozoa in Wild Cricetid Rodents from Central and Southern Chile.

In Chile, studies of parasites from the family Sarcocystidae (Apicomplexa) have mostly been related to domestic animals. We aimed to assess the presence of Sarcocystidae taxa in cricetid rodents from Central and Southern Chile. We studied 207 rodents, encompassing six species, from 13 localities. We isolated DNA from tissue samples, amplified the Sarcocystidae 18S rRNA gene with polymerase chain reaction, and performed phylogenetic analyses using maximum likelihood and Bayesian inferences. In addition, we examined blood smears and performed histological studies in organs from Sarcocystidae DNA-positive animals. Three specimens were DNA-positive and three genotypes were retrieved and named: Sarcocystis sp. P61, related to Sarcocystis strixi, was detected in two Abrothrix olivacea. Toxoplasmatinae gen. sp. P99 was retrieved from those same two specimens, and was related to Toxoplasma and other genera, although it branched independently. Besnoitia sp. R34 was detected in one Abrothrix hirta, and was clustered with congeneric species associated with rodents. No protozoa were found during microscopic studies; thus, it was not possible to confirm parasitic interactions rather than accidental encounters. However, the close relatedness of the retrieved genotypes to parasites of rodents supports the hypothesis of host-parasite associations. All three genotypes are suggested as potential new taxa, including a putative new genus.

A Novel Relapsing Fever Group Borrelia Isolated from Ornithodoros Ticks of the Brazilian Caatinga.

Tick-borne relapsing fever group (RFG) borreliosis remains neglected as a human disease and little is known on its maintenance in ticks and vertebrates, especially in South America. Therefore, this study investigated borrelial infection in Ornithodoros ticks collected in rodent-inhabited rock formations in the Brazilian semiarid region, within the Caatinga biome. Collected ticks (Ornithodoros rietcorreai and Ornithodoros cf. tabajara) were allowed to feed under laboratory conditions on guinea pigs, which had blood samples examined daily by dark-field microscopy. No spirochetes were visualized in the blood of any of four O. rietcorreai-infested guinea pigs. Contrastingly, spirochetes were visualized between 9 and 39 days after tick feeding in the blood of three guinea pigs, each infested with O. cf. tabajara ticks from a different locality. Guinea pig infection was confirmed by passages into experimental animals and by generating DNA sequences of Borrelia spp. from the blood of spirochetemic guinea pigs. Three O. cf. tabajara populations were infected by the same borrelial organism, which was characterized as a novel RFG agent (named as 'Candidatus Borrelia caatinga') based on 10 Borrelia loci (rrs, flaB, glpQ, gyrB, clpX, pepX, pyrG, recG, rplB and uvrA). We demonstrated that O. cf. tabajara is a competent vector of the novel Borrelia sp. isolates, although none of the infected rodents developed clinical illness.

Wild deer (Pudu puda) from Chile harbor a novel ecotype of Anaplasma phagocytophilum.

BACKGROUND: Deer species play an important role in the enzootic cycles of several Anaplasma species. While in the Northern Hemisphere ticks of genus Ixodes are well recognized vectors of these intracellular bacteria, less is known regarding the biological cycles of Anaplasma spp. in South America. METHODS: Using PCR protocols and Sanger sequencing, we assessed the presence of Anaplasma spp. in blood and ticks collected on a native deer species (Pudu puda) from southern Chile. RESULTS: Based on phylogenetic analyses of the 16S rRNA, gltA and groEL genes and calculation of average sequence divergence for groEL, our results bring to light a novel genovariant of Anaplasma phagocytophilum (named strain "Patagonia"). The strain represents a novel ecotype within the A. phagocytophilum species complex and was detected in both P. puda and their ticks. Using a larger matrix, denser taxon sampling and outgroup, our maximum-likelihood- and Bayesian-inferred phylogenies for groEL provide an accurate picture of the topology of A. phagocytophilum ecotypes and their evolutionary relationships. CONCLUSIONS: This is the first report of an ecotype of A. phagocytophilum in South America. Our results provide novel insight into the genetic diversity and ecology of this complex of bacterial lineages. Further studies should elucidate the enzootic cycle of A. phagocytophilum strain "Patagonia" and assess its pathogenic potential for pudues, domestic animals and humans in the region.

A search for piroplasmids and spirochetes in threatened pudu (Pudu puda) and associated ticks from Southern Chile unveils a novel Babesia sp. and a variant of Borrelia chilensis.

Cervids are important hosts for ticks and although they are refractory to some tick-borne agents such as Borrelia, they do act as reservoirs for others such as Babesia. Babesia and Borrelia are commonly transmitted by Ixodes spp. associated with deer, and most of the knowledge on their biological cycles comes from northern latitudes of the globe. In this study, we performed genetic screenings to detect tick-borne agents in blood and Ixodes stilesi ticks collected from an insular population of threatened pudu (Pudu puda), a pygmy deer species that inhabits temperate rainforests of southern South America. Inferred by phylogenetic analyses for 18S rRNA, COI and cytb genes, our results unveiled a novel genospecies of Babesia (Babesia sp. pudui) genetically related to Babesia odocoilei, a species that infects Odocoileus virginianus deer in North America. Although blood of the deer was negative for Borrelia infection, multilocus sequencing typing performed in one I. stilesi tick revealed the occurrence of a novel genetic variant of Borrelia chilensis, differing 0.93% and 0.18% in flaB and pepX genes with the type of strain for the species, respectively. Such a genetic divergence could be the result of thousands of years of isolation because of recent glaciation events that separated pudus and their tick populations at Chiloé Island approximately 437,000 years ago. The finding of a Babesia sp. has no precedents for wild and domestic ungulates in Chile and shows a novel piroplasmid that must be considered now on in rehabilitation centres and zoos that attend pudu deer. Further research is now necessary to confirm pathogenic roles.

Historical overview and update on relapsing fever group Borrelia in Latin America.

Relapsing fever group Borrelia (RFGB) are motile spirochetes transmitted to mammalian or avian hosts through the bite of hematophagous arthropods, such as soft ticks (Argasidae), hard ticks (Ixodidae) and the human clothing lice. RFGB can infect pets such as dogs and cats, as well as birds, cattle and humans. Borrelia recurrentis, B. anserina and B. theileri are considered to have worldwide distribution, affecting humans, domestic birds and ruminants, respectively. Borrelia spp. associated with soft ticks are transmitted mainly by Ornithodoros ticks and thrive in endemic foci in tropical and subtropical latitudes. Nowadays, human cases of soft tick-borne relapsing fever remain neglected diseases in several countries, and the impact these spirochetes have on the health of wild and domestic animals is largely understudied. Human infection with RFGB is difficult to diagnose, given the lack of distinguishing clinical features (undifferentiated febrile illness). Clinically, soft tick or louse-borne relapsing fever is often confused with other etiologies, such as malaria, typhoid or dengue. In Latin America, during the first half of the twentieth century historical documents elaborated by enlightened physicians were seminal, and resulted in the identification of RFGB and their associated vectors in countries such as Mexico, Panama, Colombia, Venezuela, Peru and Argentina. Almost 80 years later, research on relapsing fever spirochetes is emerging once again in Latin America, with molecular characterizations and isolations of novel RFGB members in Panama, Bolivia, Brazil and Chile. In this review we summarize historical aspects of RFGB in Latin America and provide an update on the current scenario regarding these pathogens in the region. To accomplish this, we conducted an exhaustive search of all the published literature for the region, including old medical theses deposited in libraries of medical academies. RFGB were once common pathogens in Latin America, and although unnoticed for many years, they are currently the focus of interest among the scientific community. A One Health perspective should be adopted to tackle the diseases caused by RFGB, since these spirochetes have never disappeared and the maladies they cause may be confused with etiologies with similar symptoms that prevail in the region.

Morphological, Behavioral, and Molecular Characterization of Avian Schistosomes (Digenea: Schistosomatidae) in the Native Snail Chilina dombeyana (Chilinidae) from Southern Chile.

Avian schistosomes are blood flukes parasitizing aquatic birds and snails, which are responsible for a zoonotic disease known as cercarial dermatitis, a hypersensitive reaction associated to the cutaneous penetration of furcocercariae. Despite its worldwide distribution, its knowledge is fragmentary in the Neotropics, with most of data coming from Argentina and Brazil. In Chile, there are only two mentions of these parasites from birds, and one human outbreak was associated to the genus "Trichobilharzia". However, the identity of such parasites is pending. The aim of this study was to identify the furcocercariae of avian schistosomes from Southern Chile using an integrative approach. Thus, a total of 2283 freshwater snails from different families were collected from three different regions. All snails were stimulated for the shedding of furcocercariae, but only Chilina dombeyana (Chilinidae) from the Biobío region was found to be parasitized. The morphology and phylogenetic analyses of 28S and COI genes stated two lineages, different from Trichobilharzia, shared with Argentina. This study provides new information on Neotropical schistosomes, highlighting the need for major research on these neglected trematodes, which are considered to be emerging/re-emerging parasites in other parts of the globe as consequence of anthropogenic disturbances and climatic change. Highlights: 1. Two different lineages (Lineage I and II) were described and molecularly characterized (28S and COI genes); 2. Cercaria chilinae I y II are proposed as a synonymous of Lineage II. Thus, a total of four different lineages of avian schistosomes are related to Chilina spp.; 3. Chilina spp. represents an important intermediate host for avian schistosomes in South America, constituting a reservoir de schistosomes with zoonotic potential; 4. Coinfection between the two different lineages was found, a finding previously not reported for avian schistosomes; 5. Expansion in the geographic distribution of Nasusbilharzia melancorhypha from its original record in Argentina, with Chilina dombeyana as an additional intermediate host.

Apicomplexans in small mammals from Chile, with the first report of the Babesia microti group in South American rodents.

Small mammals play an essential role as disseminators of pathogens because they reach high population densities and have ubiquitous distributions. In the Northern Hemisphere rodents are well recognized as reservoirs for tick-borne bacteria of the Anaplasmataceae family and also apicomplexan protozoans. In contrast, South American rodents hosting these microorganisms have been rarely identified. In this study, we collected blood from rodents and marsupials in northern Chile and screened for Anaplasmataceae bacteria and apicomplexan protozoa. Overall, 14.7% of the samples were positive for Babesia, Hepatozoon, and Sarcocystidae using conventional PCR assays targeting the structural 18S rRNA locus (18S). Phylogenetic analyses performed with amplicons derived from 18S and cytochrome c oxidase (COI) gene provided evidence of a Babesia sp. belonging to the Babesia microti group in Phyllotis darwini, and a novel Babesia genotype in P. darwini and Abrothrix jelskii. Furthermore, four novel genotypes of Hepatozoon retrieved from Abrothrix olivacea, P. darwini, and Oligoryzomys longicaudatus, formed independent lineages within a clade that includes additional Hepatozoon spp. detected in South American rodents. Moreover, an incidental finding of a previously detected apicomplexan, herein designated as Sarcocystidae sp., was recorded in Thylamys opossums with a high prevalence, indicating a possible specific association with these mammals. Phylogenetic analysis of Sarcoystidae sp. clearly demonstrated its relatedness to apicomplexans detected in Australian marsupials. Our results expand the range of mammals hosting tick-borne apicomplexans in South America, highlight a novel clade consisting of South American babesias, and report for the first time the B. microti group infecting rodents in the region.

Trichinella spiralis in a cougar (Puma concolor) hunted by poachers in Chile.

Trichinellosis is a zoonosis caused by nematodes of the Trichinella genus, of which 10 species have been described. Species identification when larvae is found is only possible using molecular tools. Meat from pigs and game animals not subjected to veterinary inspection are the main sources of human infections. The hunting of native carnivores is prohibited in Chile due to conservation issues and the fact that those animals favor pest control. The illegal hunting of a cougar (Puma concolor) occurred in September 2020. Herein, the molecular identification of Trichinella larvae, by analyzing nuclear (expansion segment V) and mitochondrial (cytochrome C oxidase subunit I) sequences are described. Both the amplification of the expansion segment V region and the phylogenetic analysis of a segment of a fragment of the cytochrome c-oxidase subunit I sequence confirmed that the larvae belonged to T. spiralis. The case described herein represents the first evidence of illegal hunting of a protected mammal infected with Trichinella in Chile, highlighting the 'One Health' perspective to face this disease in the rural-sylvatic interphase.

Trichinella spiralis in a cougar (Puma concolor) hunted by poachers in Chile / Trichinella spiralis em um puma (Puma concolor) caçado por caçadores furtivos no Chile

Abstract Trichinellosis is a zoonosis caused by nematodes of the Trichinella genus, of which 10 species have been described. Species identification when larvae is found is only possible using molecular tools. Meat from pigs and game animals not subjected to veterinary inspection are the main sources of human infections. The hunting of native carnivores is prohibited in Chile due to conservation issues and the fact that those animals favor pest control. The illegal hunting of a cougar (Puma concolor) occurred in September 2020. Herein, the molecular identification of Trichinella larvae, by analyzing nuclear (expansion segment V) and mitochondrial (cytochrome C oxidase subunit I) sequences are described. Both the amplification of the expansion segment V region and the phylogenetic analysis of a segment of a fragment of the cytochrome c-oxidase subunit I sequence confirmed that the larvae belonged to T. spiralis. The case described herein represents the first evidence of illegal hunting of a protected mammal infected with Trichinella in Chile, highlighting the 'One Health' perspective to face this disease in the rural-sylvatic interphase.

Resumo A triquinelose é uma doença causada por nematoides do gênero Trichinella. Dez espécies foram descritas. A identificação das espécies a partir das larvas só foi possível com ferramentas moleculares. O abate doméstico de suínos e o consumo de animais de caça, sem inspeção veterinária, são as principais fontes de infecção humana. A caça de carnívoros nativos é proibida no Chile por questões de conservação e pelo fato de que esses animais favorecem o controle de pragas. A caça ilegal de um puma (Puma concolor) aconteceu em setembro de 2020. No Chile, a identificação molecular de larvas de Trichinella, obtida pelo exame de sequências nucleares (segmento de expansão V) e mitocondriais (citocromo C oxidase subunidade I) são descritas. Tanto a amplificação da região do segmento de expansão V quanto a análise filogenética de um segmento de um fragmento da sequência da subunidade I da Citocromo c-oxidase, confirmaram que as larvas pertenciam a T. spiralis. O caso aqui descrito representa a primeira evidência de caça ilegal de um mamífero protegido infectado com Trichinella no Chile, destacando a perspectiva de "Saúde Única" para enfrentar esta doença na interfase rural-silvestre.

Rodents as potential reservoirs for Borrelia spp. in northern Chile.

Small mammals play an essential role in the transmission and maintenance cycles of Borrelia spirochetes. In Chile, recent studies have characterized novel Borrelia genotypes in ticks collected from small mammals, a fact that suggests these vertebrates are hosts for spirochetes from this genus. Considering this evidence, the goal of this study was to determine the presence of Borrelia DNA in small mammals inhabiting northern Chile. In winter of 2018, 58 small mammals were captured in five localities. Blood samples were collected from rodents and DNA was extracted to determine the presence of Borrelia DNA by PCR targeting the flaB gene and rrs-rrlA intergenic spacer (IGS). From three individuals (5%), belonging to two rodent species of Cricetidae family (Phyllotis xanthopygus and Oligoryzomys longicaudatus), we retrieved three flaB and two IGS Borrelia genotypes. Phylogenetic analyses performed with both Maximum Likelihood and Bayesian inferences showed that our sequences grouped with homologous genotypes from the relapsing fever and Lyme borreliosis groups. Our findings suggest that P. xanthopygus and O. longicaudatus rodents may play a role as reservoirs for borrelial spirochetes in Chile.

Molecular detection of Rickettsia spp., Anaplasma platys and Theileria equi in ticks collected from horses in Tayrona National Park, Colombia.

Horses are among the domestic animals that closely interact with humans and are highly parasitized by ticks, which are the primary vectors of zoonoses. As horses in Tayrona National Natural Park (PNNT) are used as a means of transporting goods, luggage and people, they are in constant contact with wild animals, workers and tourists from different countries. These factors increase the transmission risk of hemoparasites. The purpose of this study was to determine the presence of Rickettsia sp., Anaplasma sp., and Theileria sp., in horse ticks in this protected area using conventional PCR. We collected 343 ticks of genera Amblyomma, Rhipicephalus and Dermacentor. Of the 61 samples analyzed by PCR, 18 (29.5%) individuals were positive for Rickettsia sp., 15 (24.5%) for Anaplasma sp. and 4 (6.6%) for Theileria sp. This is the first report of these hemoparasite genera in ticks associated with horses in this preserved natural area, demonstrating the importance of additional studies on the presence and epidemiology of hemoparasites and their vectors in domestic and wild animals in conserved areas with a high flow of tourists.

Hemogregarine and Rickettsial infection in ticks of toads from northeastern Colombia.

The toads Rhinella spp. are in constant contact with humans and domestic animals and are commonly parasitized by ticks, which are also potential vectors of pathogenic microorganisms, such as apicomplexans and rickettsia. However, little is known about microorganisms associated with toad ticks. In this work, we molecularly evaluated the presence of Rickettsia spp. and hemogregarines in ticks of Rhinella horribilis and R. humboldti in the Colombian Caribbean, finding two different species of Rickettsia: the colombianensi strain and one close to R. bellii. In the case of hemogregarines, since only 18S gene sequences are available, it is difficult to define species and place them correctly in a phylogeny, but most of our samples show a 99% identity with Hemolivia stellata, while others identical to each other seem to form another clade within this genre. All collected ticks were identified as Amblyomma dissimile, representing the first time that H. stellata was recorded in this tick. The prevalence of both microorganisms was very high, which makes it necessary to generate robust phylogenies to clarify their taxonomic diversity and to correctly define their ecological role and pathogenicity, which should be taken into account in amphibian conservation plans and veterinary medicine.

Rickettsial infection in ticks (Acari: Ixodidae) from reptiles in the Colombian Caribbean.

Although more reptiles are illegally traded in Colombia than any other group of animals, for both local and international markets, little is known about ticks associated with reptiles or pathogens associated with these ticks. In this study, ticks were collected from reptiles in Magdalena, Cesar and La Guajira regions in northern Colombia, and identified morphologically using taxonomic keys and molecularly by sequencing of the COI gene. In addition, Rickettsia spp. were detected by PCR amplifying the gltA, 16S rRNA, and sca1 genes. Ticks were identified as Amblyomma dissimile, representing the first record for this species in 17 of the 26 species of reptiles sampled. Additionally, our data expand the distribution range for this tick in Colombia. Rickettsia spp. DNA was detected in 18 ticks (3.6%) from 16 reptiles. Sequence analyses indicated that most of the Rickettsia were members of the R. monacensis clade, most closely related to Rickettsia sp. strain Colombianensi. We detected one tick infected with a Rickettsia with 99% identity to R. bellii.