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.

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.

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.

Seroprevalence and Genotypic Analysis of Ehrlichia canis Infection in Dogs and Humans in Cauca, Colombia.

Ehrlichia canis infections have been reported in humans in Venezuela and Costa Rica. In this study, 506 healthy residents and 114 dogs from four municipalities (Cauca, Colombia) were surveyed and blood samples collected. Antibodies to E. canis in human and canine sera were evaluated using the Tandem repeat protein 19 (TRP19) peptide ELISA and indirect immunofluorescence assay (IFA). Ehrlichia canis TRP19 antibodies were detected in only 1/506 human sera, but the single positive sample was negative by IFA. The majority (75/114; 66%) of dogs surveyed had antibodies to the E. canis TRP19 peptide by ELISA, and eight randomly selected sera were further confirmed by E. canis IFA. Genomic DNA samples obtained from 73 E. canis TRP19 ELISA-positive dog blood samples were examined by PCR targeting the 16S ribosomal ribonucleic acid (rRNA) gene. Ehrlichia canis 16S rRNA was amplified in 30 (41%) of the dogs, and 16 amplicons were selected for DNA sequencing, which confirmed that all were E. canis. A second PCR was performed on the 16 confirmed E. canis 16S rRNA PCR-positive samples to determine the TRP36 genotype by amplifying the trp36 gene. TRP36 PCR amplicon sequencing identified nine dogs infected with the U.S. E. canis TRP36 genotype (56%), one dog with the Brazilian genotype (6%), and six dogs with the Costa Rican genotype (38%). Moreover, these molecular genotype signatures were consistent with serologic analysis using TRP36 genotype-specific peptides. Notably, there was no serologic evidence of E. canis infection in humans, suggesting that E. canis infection in dogs in Cauca is not associated with zoonotic human infection.

Candidatus Rickettsia senegalensis in Cat Fleas (Siphonaptera: Pulicidae) Collected from Dogs and Cats in Cauca, Colombia.

Rickettsia typhi and Rickettsia felis (Rickettsiales: Rickettsiaceae) are flea-transmitted pathogens. They are important causes of acute febrile illness throughout the world. We, therefore, sought to identify the rickettsial species present in the fleas of dogs and cats in the department of Cauca, Colombia. In this study, we collected 1,242 fleas from 132 dogs and 43 fleas from 11 cats. All fleas were morphologically identified as Ctenocephalides felis (Bouché) adults and organized in pools for DNA extraction (234 pools from dogs and 11 from cats). The gltA gene from rickettsiae was targeted for screening amplification using conventional PCR. In total, 144 of the 245 pools (58.7%) were positive. The positive samples were then processed for the amplification of the 17kDa antigen gene (144/144; 100% positive) and sca5 gene (140/144; 97.2% positive). In addition, restriction enzyme length polymorphism analysis using NlaIV on the amplified product of the sca5 gene demonstrated several organisms: 21/140 (15%) were R. felis, 118/140 (84.3%) were Rickettsia asemboensis, and 1/140 (0.7%) were Candidatus Rickettsia senegalensis. Subsequent sequencing confirmed Candidatus Rickettsia senegalensis in C. felis collected from dogs the first reported from Colombia.

Assessing Financial Impacts of Subclinical Mastitis on Colombian Dairy Farms.

Bovine mastitis is a dairy cattle disease with high economic impact. Subclinical mastitis (SCM) contributes to most of the financial losses. Colombia dairy sector accounts for 2.3% of the gross domestic product (GDP) and 24.3% of the livestock GDP. Milk production reaches 6,500 million liters/year from nearly 500,000 cattle farms and is mainly based on small-scale production systems. This study evaluates the financial impact of SCM and the potential for its control in three dairy farm strata in a region in Colombia. The objectives of the study were 1) to determine the perception of farmers about the SCM problem on their farms, 2) to assess prevalence and financial impact of SCM on farms and in the "Area five" sanitary region of the Bogota plateau, and 3) to assess costs and effectiveness of control methods of SCM. Information about disease management and decision-making process was obtained through a participatory epidemiology workshop and applying a semi-structured survey. A two-stage stratified cross sectional epidemiological study was conducted on dairy cattle from a region with approximately 400 farms and 12,000 cows, with a sample size of 55 farms. Prevalence of SCM was calculated by defining a cow as positive for the disease when any quarter had a somatic cell count (SCC) higher than 250 × 103 cells/ml. The prevalence of SCM in cows was 55.2%; significant differences were found between strata. Assessment of the financial impact of SCM in terms of milk losses was conducted using spreadsheet models. Milk production losses per farm ranged from 1.3% to 13.5%, and the economic impact in the region was estimated over USD $800.000 per year. The financial impact was greater in small- and medium-sized farms than large farms, and it was associated with the severity of SCC per quarter. Principal component analysis showed interactions, irrespective of the individual effect, and suggested three main groups of control interventions: application of basic milking hygiene practices, increase in the level of hygiene practices and veterinary advice, and SCM diagnosis and dry-cow treatment. Lack of information on management and production at farms promotes intuitive decision-making. Further research for the deeper understanding of intervention costs and effectiveness is suggested.