Extracellular vesicles released by Trypanosoma evansi: induction analysis and proteomics.

Trypanosoma evansi is a unicellular protozoan responsible for causing a disease known as "surra," which is found in different regions of the world and primarily affects horses and camels. Few information is known about virulence factors released from the parasite within the animals. The organism can secrete extracellular vesicles (EVs), which transport a variety of molecules, including proteins. Before being considered exclusively as a means for eliminating unwanted substances, extracellular vesicles (EVs) have emerged as key players in intercellular communication, facilitating interactions between cells, host cells, and parasites, and even between parasites themselves. Thus, they may be used as potential biomarkers. This study aimed to assess the induction of EVs production by Ca+2, conduct a proteomic analysis of the EVs released by T. evansi, and identify epitopes that could serve as biomarkers. The findings indicated that Ca+2 is not an effective promoter of vesiculation in T. evansi. Furthermore, the proteomic analysis has identified multiple proteins that have been investigated as biomarkers or vaccine antigens, previously. A total of 442 proteins were identified, with 7 of them specifically recognizing 9 epitopes that are unique to T. evansi. At least one of these epitopes of TevSTIB805.9.11580 have been previously identified, which increases the possibility of further investigating its potential as a biomarker.

Electronic von Frey associated with the Mouse Grimace Scale to Assess Allodynia and Pain in Trypanosoma evansi-Infected Mice.

Infectious disease pathogenesis is still a complex field to study. The course of several clinical signs, such as allodynia and pain, may be observed in domestic animals. However, the knowledge of their pathways and correct treatment need controlled experiments, many of them using laboratory animals. Measuring changes in mechanical thresholds of the hind paw and viscera is a useful technique to observe changes in pain perception in rodents. Withdrawal response can be measured first in baseline tests, which creates better control of experimental groups. Subsequent tests can be performed after inducing infection and adding drugs to the protocol. The use of an electronic von Frey apparatus associated with the use of a facial scale to observe pain-like changes allows a simple, precise, and consistent assessment to evaluate allodynia and pain in mice. Thus, experiments using the present methodology for Trypanosoma evansi infection represent a useful method to evaluate allodynia and pain in laboratory-infected animals, which can be applied to the conventional treatment for livestock animals.

Comparative study of three novel ion exchange resins with DEAE-cellulose for the purification of Trypanosoma evansi.

Ion exchange chromatography is a method that uses the different surface charges of trypanosomes and blood cells to separate them. This makes it possible to use molecular and immunological methods to diagnose or study these protozoans. DEAE-cellulose resin is commonly used to perform this method. The goal of this study was to compare three novel chromatographic resins designated as PURIFICA™ (Y-C2N®, Y-HONOH®, and Y-CNC3®). The resins were evaluated based on their ability to isolate the parasite, purification time, examination of parasite viability and morphology, and trypanosome recovery potential after passing through the columns. In terms of the evaluated parameters, there was no significant difference between DEAE-cellulose and the three tested resins in most experiments. However, PURIFICA™ (Y-C2N®, Y-HONOH®, and Y-CNC3®) resins are less expensive and easier to prepare than DEAE-Cellulose, making them an alternative for the purification of Trypanosoma evansi.