An overview of the trypanosomatid (Kinetoplastida: Trypanosomatidae) parasites infecting several mammal species in Colombia

Abstract

Background Trypanosomatids are among the most critical parasites for public health due to their impact on human, animal, and plant health. Diseases associated with these pathogens manifest mainly in poor and vulnerable populations, where social, environmental, and biological factors modulate the case incidence and geographical distribution. Methods We used Sanger and amplicon-based next-generation sequencing (NGS) in samples from different mammals to identify trypanosomatid infections in several departments in Colombia. A total of 174 DNA samples (18 humans, 83 dogs, and 73 wild mammals) were analyzed by conventional PCR using a fragment of the heat shock protein 70 (Hsp70) gene and Sanger sequenced the positive samples. Twenty-seven samples were sent for amplicon-based NGS using the same gene fragment. Data obtained were used to perform diversity analyses. Results One hundred and thirteen samples were positive for PCR by Hsp70 fragment; these corresponded to 22.1% Leishmania spp., 18.6% L. amazonensis, 9.7% L. braziliensis, 14.2% L. infantum, 8% L. panamensis, and 27.4% Trypanosoma cruzi. Comparison of the identified species by the two sequencing technologies used resulted in 97% concordance. Alpha and beta diversity indices were significant, mainly for dogs; there was an interesting index of coinfection events in the analyzed samples: different Leishmania species and the simultaneous presence of T. cruzi and even T. rangeli in one of the samples analyzed. Moreover, a low presence of L. braziliensis was observed in samples from wild mammals. Interestingly, to our knowledge, this is the first report of Leishmania detection in Hydrochaeris hydrochaeris (capybara) in Colombia. Conclusions The Hsp70 fragment used in this study is an optimal molecular marker for trypanosomatid identification in many hosts and allows the identification of different species in the same sample when amplicon-based sequencing is used. However, the use of this fragment for molecular diagnosis through conventional PCR should be carefully interpreted because of this same capacity to identify several parasites. This point is of pivotal importance in highly endemic countries across South America because of the co-circulation of different genera from the Trypanosomatidae family. The findings show an interesting starting point for One Health approaches in which coevolution and vector-host interactions can be studied. Graphical Abstract

Validation of Oxford nanopore sequencing for improved New World Leishmania species identification via analysis of 70-kDA heat shock protein

“…Nonetheless, it is worth noting that this marker might not be efficacious in areas with a high incidence of Chagas Disease or in regions where both parasites coexist sympatrically. In such instances, the HSP70-Short marker could certainly provide higher resolution, given its capability to reliably differentiate among various trypanosomatids and consistently detect coinfection events within a single host [ 16 , 59 ].…”

Application of Next-Generation Sequencing (NGS) Techniques for Selected Companion Animals

“…have evaluated many coinfections of Trypanosomatidae in samples in which infection has been previously diagnosed by PCR [67]. Huggings et al developed and validated a longread metabarcoding platform for the detection of filarial worm pathogens of animals and humans [68].…”

“…NGS technologies could also be used to evaluate if the coinfection of few species of one pathogen occurs. Castillo-Castañeda et al have evaluated many coinfections of Trypanosomatidae in samples in which infection has been previously diagnosed by PCR [ 67 ]. Huggings et al developed and validated a long-read metabarcoding platform for the detection of filarial worm pathogens of animals and humans [ 68 ].…”

G‐Quadruplexes as Key Transcriptional Regulators in Neglected Trypanosomatid Parasites

“…They are spread worldwide but endemic in the poorest and most vulnerable populations in Central and South America, and in various regions of Africa, where treatment and prevention are often neglected [1] . These parasites are mostly transmitted by insect vectors to different hosts, including humans and both wild and domesticated animals, [2] thus presenting a serious threat to health, society, and economics [3] . Chagas disease (also known as American trypanosomiasis) is caused by the agent Trypanosoma cruzi , whilst human African trypanosomiasis (HAT; also known as African sleeping sickness) is caused by Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense , and visceral and cutaneous leishmaniasis are caused by parasites of the genus Leishmania [4] .…”