preparation and inoculation of trypanosoma evansi in mouse to evaluate allodynia and pain

Assessing Pain and Allodynia in T. evansi-Infected Animals

Trypanosoma evansi is the etiological agent of the disease called &#34;surra&#34; or &#34;mal-das-cadeiras&#34; in South America1,2. It commonly affects equines and cattle, but also wild fauna, being transmitted by hematophagous bats, tabanids, and stomoxes fly bites1,3. Surra is a major disease in domestic animals, which can be fatal in the absence of the correct treatment, exhibiting nonspecific clinical signs such as anemia, loss of appetite and weight, muscle weakness, and abortion, which may vary according to the host and geographic region2,4,5,6.
The expression of allodynia and pain in infected animals during the course of the disease is still a new topic2,7. The urge to understand the pathogenesis behind these signs is an important step to improve and refine the currently used treatment by adding effective analgesic drugs to the classic trypanocidal protocol5,6. In this scenario, the possibility of replicating the disease using a murine model represents an advantage, as mice can be easily kept under controlled environments in the laboratory and therefore, yield results that are more consistent than field experiments using livestock.
A mechanical threshold is commonly obtained using an electronic von Frey (EvF) apparatus for the evaluation of allodynia in a vast number of experiments2,8,9. This device is used to assess tissue sensitivity to mechanical stimulation: once the apparatus touches the paw of the animal, through a 20-200 &#181;L tip attached to the device, the force with which the animal withdraws the paw is recorded.
Rodents are usually used as standard animals in these experiments, and most results are extrapolated to other species, as most researched diseases come from other species in which it would be difficult to conduct a controlled study. Furthermore, allodynia and pain are intimately related. The use of a specific facial scale to evaluate pain in infected mice plays an important role as a confirming adjuvant to affirm the presence of pain during the course of T. evansi infection2,10.
In this protocol, we demonstrate a novel model to evaluate allodynia and pain in mice experimentally infected with T. evansi, showing a high correlation between variables, thus proving to be a strong model. Furthermore, it requires a small number of researchers to carry out the entire procedure, decreasing the chance of human interference during the experiment. It also enables the investigator to replicate the target disease during an acute course and add several different treatment drugs in the experimental design, thereby obtaining consistent results in a short period.

Author Spotlight: Assessing Pain and Promoting Animal Welfare in Laboratory Animals Infected with Trypanosoma evansi

Electronic von Frey forbundet med musegrimasseskalaen til vurdering af allodyni og smerte hos Trypanosoma evansi-inficerede mus

Forberedelse og podning af Trypanosoma evansi i mus for at evaluere allodyni og smerte

For at begynde skal du tænde for vandbadsudstyret og indstille det til 37 grader Celsius, fjerne et mikrocentrifugerør indeholdende det konserverede blodprøve fra ultrafryseren og placere det i vandbadet, indtil blodet tøer op. Udfør seriel fortynding ved hjælp af PBS med 60% glukose for at fortynde trypanosomerne. Tæl antallet af parasitceller ved hjælp af et neubauerkammer under et mikroskop med en 100 gange objektiv linse.

Research

JoVE Journal

Immunology and Infection

Preparation and Inoculation of Trypanosoma evansi in Mouse to Evaluate Allodynia and Pain

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 ...

Electronic Von Frey (EvF) Apparatus Set up for the Evaluation of Allodynia and Pain in Trypanosoma evansi Infected Animals

Elektronisk von Frey (EvF) apparat sat op til evaluering af allodyni og smerte hos Trypanosoma evansi-inficerede dyr

Mechanical Threshold Assessment of Mice Infected with Trypanosoma evansi

Mekanisk tærskelvurdering af mus inficeret med Trypanosoma evansi

Grimace Scale Assessment of Mice Infected with Trypanosoma evansi

Grimasseskalavurdering af mus inficeret med Trypanosoma evansi