Hyperparasitism in bat flies (Diptera: Streblidae): new records and interaction networks in the Neotropics.

Hyperparasitism is defined as the interaction where one parasite is infected by another parasite. In bat flies (Streblidae and Nycteribiidae), both hyperparasites and microparasites (bacteria, viruses, fungi, and arthropods such as mites) have been documented. Fungi belonging to the order Laboulbeniales are microscopic parasites of a wide diversity of arthropod hosts. Three genera exclusively target bat flies: Arthrorhynchus, which parasitizes species within Nycteribiidae in the Eastern Hemisphere, while genus Gloeandromyces and Nycteromyces parasitize Streblidae in the Western Hemisphere. Among the hyperparasitic arthropods, mites of family Neothrombidiidae, particularly the monospecific genus Monunguis, are known to parasitize bat flies. Here we present the first records of the hyperparasites Monunguis streblida and Gloeandromyces pageanus f. polymorphus parasitizing Streblidae bat flies in Colombia and a summary of these hyperparasitic interactions in the Neotropics. We detected fungi and mites parasitizing bat flies that were collected in the Magdalena River Basin, Colombia, in field expeditions in 2018, 2022, and 2023. We identified 17 bat flies and two species of hyperparasites, specifically M. streblida and the fungi Gloeandromyces. Our search for reports of these interactions in the Neotropics revealed that seven species of Trichobius (Streblidae) are parasitized by M. streblida, whereas Paratrichobius longicrus (Streblidae) is parasitized by Gloeandromyces pageanus f. polymorphus. These interactions have been reported in 11 countries, but our records are the first of M. streblida and Laboulbeniales fungi parasitizing bat flies in Colombia. So far, a total of 14 species of fungi and one species of mite have been associated with 19 species of bat flies, which in turn, are linked to 15 species of Neotropical bats.

Serologic Evidence of Orientia Infection among Rural Population, Cauca Department, Colombia.

We assessed serum samples collected in Cauca Department, Colombia, from 486 persons for Orientia seroreactivity. Overall, 13.8% showed reactive IgG by indirect immunofluorescence antibody assay and ELISA. Of those samples, 30% (20/67) were confirmed to be positive by Western blot, showing >1 reactive band to Orientia 56-kD or 47-kD antigens.

Emerging Tickborne Bacteria in Cattle from Colombia.

Ehrlichia minasensis is a new pathogenic bacterial species that infects cattle, and Borrelia theileri causes bovine borreliosis. We detected E. minasensis and B. theileri DNA in cattle from southwestern Colombia by using PCR. E. minasensis and B. theileri should be considered potential etiologies of febrile syndrome in cattle from Colombia.

Endothelial Cell Proteomic Response to Rickettsia conorii Infection Reveals Activation of the Janus Kinase (JAK)-Signal Transducer and Activator of Transcription (STAT)-Inferferon Stimulated Gene (ISG)15 Pathway and Reprogramming Plasma Membrane Integrin/Cadherin Signaling.

Rickettsia conorii is the etiologic agent of Mediterranean spotted fever, a re-emerging infectious disease with significant mortality. This Gram-negative, obligately intracellular pathogen is transmitted via tick bites, resulting in disseminated vascular endothelial cell infection with vascular leakage. In the infected human, Rickettsia conorii infects endothelial cells, stimulating expression of cytokines and pro-coagulant factors. However, the integrated proteomic response of human endothelial cells to R. conorii infection is not known. In this study, we performed quantitative proteomic profiling of primary human umbilical vein endothelial cells (HUVECs) with established R conorii infection versus those stimulated with endotoxin (LPS) alone. We observed differential expression of 55 proteins in HUVEC whole cell lysates. Of these, we observed induction of signal transducer and activator of transcription (STAT)1, MX dynamin-like GTPase (MX1), and ISG15 ubiquitin-like modifier, indicating activation of the JAK-STAT signaling pathway occurs in R. conorii-infected HUVECs. The down-regulated proteins included those involved in the pyrimidine and arginine biosynthetic pathways. A highly specific biotinylated cross-linking enrichment protocol was performed to identify dysregulation of 11 integral plasma membrane proteins that included up-regulated expression of a sodium/potassium transporter and down-regulation of α-actin 1. Analysis of Golgi and soluble Golgi fractions identified up-regulated proteins involved in platelet-endothelial adhesion, phospholipase activity, and IFN activity. Thirty four rickettsial proteins were identified with high confidence in the Golgi, plasma membrane, or secreted protein fractions. The host proteins associated with rickettsial infections indicate activation of interferon-STAT signaling pathways; the disruption of cellular adhesion and alteration of antigen presentation pathways in response to rickettsial infections are distinct from those produced by nonspecific LPS stimulation. These patterns of differentially expressed proteins suggest mechanisms of pathogenesis as well as methods for diagnosis and monitoring Rickettsia infections.

Bartonella quintana and Typhus Group Rickettsiae Exposure among Homeless Persons, Bogotá, Colombia.

In 2015, we investigated Bartonella quintana and typhus group rickettsiae in body lice from homeless persons in Bogotá, Colombia. We found B. quintana-infected body lice and seroprevalence of this microorganism in 19% of homeless persons and typhus group rickettsiae in 56%. Public health professionals should start preemptive measures and active vector control.

Genetic diversity of P1/pathogenic Leptospira species hosted by bats worldwide.

INTRODUCTION: Bats are a diverse group of mammals that have unique features allowing them to act as reservoir hosts for several zoonotic pathogens such as Leptospira. Leptospires have been classified into pathogenic, intermediate, and saprophytic groups and more recently into clades P1, P2, S1, and S2, being all the most important pathogenic species related to leptospirosis included within the P1/pathogenic clade. Leptospira has been detected from bats in several regions worldwide; however, the diversity of leptospires harboured by bats is still unknown. AIM: The aim of the present study was to determine the genetic diversity of Leptospira spp. harboured by bats worldwide. METHODS: A systematic review was conducted on four databases to retrieve studies in which Leptospira was detected from bats. All studies were screened to retrieve all available Leptospira spp. 16S rRNA sequences from the GenBank database and data regarding their origin. Sequences obtained were compared with each other and reference sequences of Leptospira species and analysed through phylogenetic analysis. RESULTS: A total of 418 Leptospira spp. 16S rRNA sequences isolated from 55 bat species from 14 countries were retrieved from 15 selected manuscripts. From these, 417 sequences clustered within the P1/pathogenic group, and only one sequence clustered within the P2/intermediate group. Six major clades of P1/pathogenic Leptospira spp. were identified, three of them composed exclusively of sequences obtained from bats. CONCLUSION: We identified that bats harbour a great genetic diversity of Leptospira spp. that form part of the P1/pathogenic clade, some of which are closely related to leptospirosis-associated species. This finding contributes to the knowledge of the diversity of leptospires hosted by bats worldwide and reinforces the role of bats as reservoirs of P1/pathogenic Leptospira spp.

Genetic diversity of Bartonella spp. among cave-dwelling bats from Colombia.

Bartonella is a bacterial genus that comprises arthropod-borne microorganisms. Several Bartonella isolates have been detected from bats worldwide, which are thought to be undescribed species. We aimed to test the presence of Bartonella spp. among bats from Colombia, and evaluate the genetic diversity of bat-associated Bartonella spp. through phylogenetic analyses. A total of 108 bat blood samples were collected from three bat species (Carollia perspicillata, Mormoops megalophylla, and Natalus tumidirostris) that inhabit the Macaregua cave. The Bartonella ssrA gene was targeted through real-time and end-point PCR; additionally, the gltA and rpoB genes were detected by end-point PCR. All obtained amplicons were purified and bidirectionally sequenced for phylogenetic analysis using a concatenated supermatrix and a supertree approaches. A detection frequency of 49.1 % (53/108) for Bartonella spp. was evidenced among bat blood samples, of which 59.1 % (26/44), 54.3 % (19/35) and 27.6 % (8/29) were identified in Carollia perspicillata, Natalus tumidirostris and Mormoops megalophylla respectively. A total of 35 ssrA, 5 gltA and 4 rpoB good-quality sequences were obtained which were used for phylogenetic analysis. All obtained bat sequences clustered together with sequences obtained from Neotropical bat species into two bat-restricted clades namely clade A and clade N. We detected the presence of Bartonella spp. that clustered within two different bat-associated Bartonella clades, giving the first data of the genetic diversity of these bacteria among bats from Colombia.

Molecular Characterization of Leptospira Species among Patients with Acute Undifferentiated Febrile Illness from the Municipality of Villeta, Colombia.

Leptospira is a bacterial genus that includes several pathogenic species related to leptospirosis. In Colombia, leptospirosis is a mandatorily reported disease, widely distributed across the country. In the Villeta municipality, leptospirosis has been identified as an important cause of febrile illness; however, to date, no studies have been performed to identify the circulating species. A genus-specific qualitative qPCR was performed on DNA extracted from febrile patients' acute-phase whole-blood samples targeting a fragment of the rrs gene. Positive qPCR samples were further amplified for the adk, icdA, LipL32, LipL41, rrs, and secY genes through conventional PCR for sequencing. All high-quality obtained sequences were further assessed through concatenated phylogenetic analysis. A total of 25% (14/56) of febrile patients' acute blood samples were positive for Leptospira spp. High-quality sequences were obtained for only five genes, and analysis through concatenated phylogeny identified that all sequences clustered within the P1/pathogenic clade; some of them formed a robustly supported clade with Leptospira santarosai, and others were closely related with other Leptospira species but exhibited considerable genetic divergence. We describe the presence of pathogenic Leptospira species among febrile patients from the Villeta municipality and identify L. santarosai and other Leptospira species as causative agents of leptospirosis in the region.

Establishment of a multisite umbrella cohort study protocol to describe the epidemiology and aetiologies of acute undifferentiated febrile illness in Latin America.

INTRODUCTION: Acute undifferentiated febrile illnesses (AUFIs) impose a large burden in the tropics. Understanding of AUFI's epidemiology is limited. Insufficient diagnostic capacity hinders the detection of outbreaks. The lack of interconnection in healthcare systems hinders timely response. We describe a protocol to study the epidemiology and aetiologies of AUFI and pathogen discovery in strategic areas of Latin America (LA). METHODS AND ANALYSIS: Global Infectious Diseases Network investigators comprising institutions in Colombia, Dominican Republic, México, Perú and the USA, developed a common cohort study protocol. The primary objective is to determine the aetiologies of AUFI at healthcare facilities in high-risk areas. Data collection and laboratory testing for viral, bacterial and parasitic agents are performed in rural and urban healthcare facilities and partner laboratories. Centralised laboratory and data management cores deploy diagnostic tests and data management tools. Subjects >6 years with fever for <8 days without localised infection are included in the cohort. They are evaluated during the acute and convalescent phases of illness. Study personnel collect clinical and epidemiological information. Blood, urine, nasal or pharyngeal swabs and saliva are collected in the acute phase and blood in convalescent phase. Specimens are banked at -80°C. Malaria, dengue and COVID-19 are tested onsite in the acute phase. The acute-phase serum is PCR tested for dengue, chikungunya, Venezuelan equine encephalitis, Mayaro, Oropouche, Zika, and yellow fever viruses. Paired convalescent and acute serum antibody titters are tested for arbovirus, Leptospira spp, and Rickettsia spp. Serum is used for viral cultures and next-generation sequencing for pathogen discovery. Analysis includes variable distributions, risk factors and regression models. Laboratory results are shared with health authorities and network members. ETHICS AND DISSEMINATION: The protocol was approved by local ethics committees and health authorities. The results will be published in peer-reviewed journals. All study results are shared with local and regional health authorities.

Etiologies of Zoonotic Tropical Febrile Illnesses That Are Not Part of the Notifiable Diseases in Colombia.

In Colombia, tropical febrile illnesses represent one of the most important causes of clinical attention. Febrile illnesses in the tropics are mainly zoonotic and have a broad etiology. The Colombian surveillance system monitors some notifiable diseases. However, several etiologies are not monitored by this system. In the present review, we describe eleven different etiologies of zoonotic tropical febrile illnesses that are not monitored by the Colombian surveillance system but have scientific, historical, and contemporary data that confirm or suggest their presence in different regions of the country: Anaplasma, Arenavirus, Bartonella, relapsing fever group Borrelia, Coxiella burnetii, Ehrlichia, Hantavirus, Mayaro virus, Orientia, Oropouche virus, and Rickettsia. These could generate a risk for the local population, travelers, and immigrants, due to which they should be included in the mandatory notification system, considering their importance for Colombian public health.

Zoonotic febrile illnesses misdiagnosed as COVID-19: a review of reported clinical cases.

COVID-19 is a zoonotic coronavirus disease caused by SARS-CoV-2. Its fast spreading by aerosol transmission has made it a highly contagious disease, causing the most recent 2020 pandemic. Although it mainly affects the respiratory system, atypical forms of the disease have been described, including developing an undifferentiated febrile illness without respiratory symptoms, that can represent a diagnostic challenge, mainly in tropical areas where several zoonotic febrile diseases are circulating. Thus, despite the broad clinical spectrum of COVID-19, in the tropics, other zoonotic etiologies should always be considered as differential diagnoses. According to our case reports review, eight different zoonotic febrile diseases misdiagnosed as COVID-19 have been reported in the available scientific literature of four databases. These were only suspected due to the epidemiological history. Thus, making a complete and detailed clinical history of a febrile patient in the tropics is essential to suspect the etiology and request the necessary confirmatory tests. Therefore, COVID-19 must be included as a differential diagnosis of undifferentiated febrile illness in the tropics, but other zoonotic infectious diseases must not be ruled out.

Etiological characterization of acute undifferentiated febrile illness in Apartadó and Villeta municipalities, Colombia, during COVID-19 pandemic.

Background: Acute undifferentiated febrile illness (AUFI) is one of the leading causes of illness in tropical regions. Although malaria is the most important cause, other pathogens such as Dengue (DENV), Leptospira and recently, Coronavirus Disease 2019 (COVID-19) have gained importance. In Colombia, few studies aimed to identify the etiology of AUFI. Most of them performed in Apartadó and Villeta municipalities, identifying the active circulation of several pathogens. Thus, we conducted a cross-sectional study in these municipalities to characterize the etiologies of AUFI during COVID-19 pandemic. Methods: An active surveillance was conducted between September and December 2021 in local hospitals of Apartadó and Villeta municipalities. Febrile patients were enrolled after voluntarily agreeing to participate in the study. Ten different etiologies were evaluated through direct, serological, molecular and rapid diagnostic methods. Results: In Apartadó a confirmed etiology was found in 60% of subjects, DENV (25%) being the most frequent, followed by leptospirosis (16.7%), malaria (10%), COVID-19 (8.3%), spotted fever group (SFG) rickettsiosis (6.7%) and Chikungunya (1.7%). In Villeta, a specific etiology was confirmed in 55.4% of patients, of which SFG rickettsiosis (39.3%) was the most frequent, followed by leptospirosis (21.4%), DENV (3.6%) and malaria (1.8%). No cases due to Mayaro, Yellow Fever, Oropouche and Venezuelan Equine Encephalitis viruses were detected. Conclusion: We confirm the relevance of dengue fever, leptospirosis, SFG rickettsiosis, COVID-19 and malaria as causes of AUFI in the municipality of Apartadó, and highlight the great importance of SFG rickettsiosis as the main cause of AUFI in the municipality of Villeta.

Molecular detection of tick-borne rickettsial pathogens in ticks collected from domestic animals from Cauca, Colombia.

Some hard ticks' species can act as vectors of a wide variety of pathogens of human and animal importance such as Anaplasma, Ehrlichia and Rickettsia spp. In Colombia, a total of forty-six tick species have been described, and some of them have been implicated as vectors of some infectious agents. The department of Cauca is one of the thirty-two departments of Colombia. Most of its population lives in rural areas and depends on agriculture as the main economic activity, favoring exposure to ticks and tick-borne pathogens. Thus, the present study aimed to determine the tick species and tick-borne pathogens circulating in this region. From August to November 2017, ticks were collected from dogs, horses and cattle from eight rural areas of four municipalities in the department of Cauca. All collected ticks were classified according to taxonomic keys and organized in pools. DNA was extracted from all tick pools for molecular confirmation of tick species and detection of Anaplasma, Ehrlichia and Rickettsia spp. A total of 2809 ticks were collected which were grouped in 602 pools. Ticks were morphologically identified as Amblyomma cajennense sensu lato, Dermacentor nitens, Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato. The molecular identity of A. cajennense s.l. was confirmed as Amblyomma patinoi. A total of 95% of the pools scored positive for members of the Anaplasmataceae family, of which, 7.8% and 7.3% were positive to Anaplasma and Ehrlichia spp., respectively, being identified as Anaplasma marginale, Ehrlichia minasensis and Ehrlichia canis; and 16.1% were positive for Rickettsia spp. with high identity for Rickettsia asembonensis, Rickettsia felis and Candidatus Rickettsia senegalensis. This is the first report describing the natural infection of ticks with rickettsial pathogens and the occurrence of A. patinoi ticks in Cauca department, Colombia.

First molecular evidence of Coxiella burnetii in bats from Colombia.

Coxiella burnetii is the etiologic agent of Q fever, a zoonotic infectious disease of worldwide distribution that has a wide clinical spectrum. Transmission of C. burnetii occurs by inhalation of contaminated secretions and excreta of infected animal species, particularly goats, cattle and sheep. Activities associated with livestock contact represent the principal risk factor, however participation of wildlife reservoirs is underestimated. Thus, the aim of this study was to evaluate the presence of C. burnetii DNA in blood from bats. Molecular analyses using a qPCR targeting the IS1111 specific gene to detect DNA of C. burnetii in blood samples from 126 bats captured in the Macaregua cave, Colombia, between 2014, 2015 and 2018 were performed. Molecular evidence of C. burnetii was found in 6.3%. Results obtained in the present study represent the first detection of C. burnetii among bats in Colombia, suggesting that more studies need to be done in order to determine the role of these animals in the eco-epidemiology of Q fever.

Amblyomma mixtum free-living stages: Inferences on dry and wet seasons use, preference, and niche width in an agroecosystem (Yopal, Casanare, Colombia).

The formulation of effective control strategies for any pest species generally involves the study of habitat use and preference and niche width in anthropogenically transformed natural landscapes. We evaluated whether the use, habitat preference, and niche range of the Amblyomma mixtum tick changed between stages, habitats, and seasonality (dry-wet seasons 2019) on a farm in Yopal (Casanare, Colombia). The presence and relative abundance of free-living larvae, nymphs, and adults was quantified in four different habitats according to the type of vegetation cover (Riparian Forest, Cocoa Crop, King Grass Crop, and Star Grass Paddock). Habitat availability was estimated, environmental variables were analyzed, and various indices of habitat use and preference, and niche width were calculated. A. mixtum's habitat use and preference, and niche width changed between stages, habitat types, and time of the year. The total abundance of A. mixtum was an order of magnitude greater in the dry season than the wet season. In the dry season, all stages used all habitats, while A. mixtum adults used all the habitats in both seasons. In the dry season, nymphs and larvae preferred three out of the four habitats, while adults preferred the King Grass Crop. In the wet season, nymphs and larvae preferred two habitats, whereas the adults preferred the King Grass Crop. The value of the niche width index was high for larvae, nymphs, and adults in the dry season, while it was high only for adults in the wet season. Thus, A. mixtum's vast environmental tolerance and niche breadth allows the species to use and colonize changing habitats (unstable or temporary) with fluctuating environmental conditions (e.g., King Grass Crop), potentially keeping a stable population over time and making it an extremely resistant species. However, the wet flooding season in Yopal may exceed A. mixtum's stages' tolerances.

Pathogenic Leptospira Species in Bats: Molecular Detection in a Colombian Cave.

Leptospirosis is caused by pathogenic Leptospira spp., which can be found in nature among domestic and wild animals. In Colombia, the Macaregua cave is known for its bat richness; thus, because bats are reservoir hosts of human microbiological pathogens, we determined if the Macaregua cave bats harbored Leptospira in the wild. A total of 85 kidney samples were collected from three bat species (Carollia perspicillata, Mormoops megalophylla, and Natalus tumidirostris) to detect Leptospira spp. The 16S rRNA gene was targeted through conventional PCR and qPCR; in addition, the LipL32 gene was detected using conventional PCR. Obtained amplicons were purified and sequenced for phylogenetic analysis. The Leptospira spp. 16S rRNA gene was detected in 51.8% bat kidneys, of which 35 sequences were obtained, all clustering within the pathogenic group. Moreover, 11 sequences presented high-identity-values with Leptospiranoguchii, Leptospiraalexanderi, Leptospiraborgpetersenii, Leptospirakirschneri, and Leptospiramayottensis. From the 16S rRNALeptospira spp.-positive population samples, 28 amplified for the LipL32 gene, and 23 sequences clustered in five different phylogenetic groups. In conclusion, we detected the circulation of different groups of Leptospira spp. sequences among cave bats in the wild; some sequences were detected in more than one bat specimen from the same species, suggesting a conspecific transmission within the cave.

Arbovirus Antibody Seroprevalence in the Human Population from Cauca, Colombia.

Several arboviruses have emerged or reemerged into the New World during the past several decades, causing outbreaks of significant proportion. In particular, the outbreaks of Dengue virus (DENV), Zika virus, and Chikungunya virus (CHIKV) have been explosive and unpredictable, and have led to significant adverse health effects. These viruses are considered the leading cause of acute undifferentiated febrile illnesses in Colombia. However, Venezuelan equine encephalitis virus (VEEV) is endemic in Colombia, and arboviruses such as the Mayaro virus (MAYV) and the Oropouche virus (OROV) cause febrile illnesses in neighboring countries. Yet, evidence of human exposure to MAYV and OROV in Colombia is scarce. In this study, we conducted a serosurvey study in healthy individuals from the Cauca Department in Colombia. We assessed the seroprevalence of antibodies against multiple arboviruses, including DENV serotype 2, CHIKV, VEEV, MAYV, and OROV. Based on serological analyses, we found that the overall seroprevalence for DENV serotype 2 was 30%, 1% for MAYV, 2.6% for CHIKV, 4.4% for VEEV, and 2% for OROV. This study provides evidence about the circulation of MAYV and OROV in Colombia, and suggests that they-along with VEEV and CHIKV-might be responsible for cases of acute undifferentiated febrile illnesses that remain undiagnosed in the region. The study results also highlight the need to strengthen surveillance programs to identify outbreaks caused by these and other vector-borne pathogens.

Outbreak of Rocky Mountain spotted fever in Córdoba, Colombia.

Rocky Mountain spotted fever (RMSF) is a tick-borne disease caused by the obligate intracellular bacterium Rickettsia rickettsii. Although RMSF was first reported in Colombia in 1937, it remains a neglected disease. Herein, we describe the investigation of a large cluster of cases of spotted fever rickettsiosis in a new area of Colombia.

Clinical, epidemiological, and laboratory features of Rickettsia parkeri rickettsiosis: A systematic review.

Rickettsia parkeri rickettsiosis is recognized as the second most prevalent tick-borne disease caused by spotted fever group rickettsiae in the Americas, where two pathogenic strains (R. parkeri sensu stricto and R. parkeri strain Atlantic rainforest) have been related to human infections and transmitted by Amblyomma spp. ticks. We developed a systematic review that evaluated all available evidence in the literature regarding clinical, epidemiological, and laboratory features of R. parkeri rickettsiosis, including confirmed and probable cases. We followed the recommendations made by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guide. We excluded papers that contained missing information of some variables and publications in which it was not possible to separate data for confirmed and probable cases. A total of 77 clinical cases (32 confirmed cases and 45 probable cases) were considered for this review. Overall, our results show that R. parkeri rickettsiosis is more frequent in males in the age group of 18-64 years and that a history of tick exposure was frequent (>90%). Cases were described in the United States, Argentina, Brazil, Uruguay and Colombia. Clinically, more than 60% of the cases had fever (mean of 93%), eschar (mean of 87%), and rash (mean of 68%). Headache and myalgia were predominant nonspecific symptoms (mean of 67% and 61%, respectively). Our results show that at least 60% of R. parkeri cases had altered laboratory parameters, most often showing an increase in transaminases and leukopenia. Tetracyclines-class antibiotics were used in most (>85%) of the patients. Overall, only 9% of cases required hospitalization and there was a 100% rate of clinical recovery in all of cases.

Experimental infection and vector competence of Amblyomma patinoi, a member of the Amblyomma cajennense species complex, for the human pathogen Rickettsia rickettsii.

Amblyomma patinoi ticks infected with Rickettsia rickettsii are present in Colombia, but its vector competence is unknown. Hence, we evaluated the vector competence of A. patinoi with R. rickettsii under laboratory conditions. Experimental guinea pigs and rabbits (males and females) were separated in the infected group (IG) and the control group (CG). In the IG, the filial 1 (F1) larvae (R. rickettsii-free) from Colombian A. patinoi engorged female specimens were exposed to R. rickettsii (ITU strain) by feeding on infected guinea pigs. Next, F1 nymphs and adults, and F2 larvae were allowed to feed on uninfected guinea pigs or rabbits and tested by qPCR targeting the gltA rickettsial gene. All animals used to feed the IG F1 ticks became febrile and had R. rickettsii infection (89% fatality rate) detected through serological or molecular techniques. After the F1 larvae ticks became R. rickettsii infected, subsequent IG tick stages were able to maintain the rickettsial infection by transstadial maintenance to all infested animals, indicating A. patinoi vector competence. Subsequently, almost 31% of the F1 female egg masses and only 42% of their F2 larvae were infected. Less than 50% of the infected females transmitted R. rickettsii transovarially, and only a part of the offspring were infected. This study demonstrated that A. patinoi might not be able to sustain R. rickettsii infection by transovarial transmission for successive tick generations without horizontal transmission via rickettsemic hosts. This condition might result in low R. rickettsii-infection rates of A. patinoi under natural conditions.