In vitro production of Trypanosoma equiperdum antigen and its evaluation for use in serodiagnosis of dourine

Bassarak, Björn; Moser, Irmgard; Menge, Christian

doi:10.1016/j.vetpar.2016.04.032

Cited by 4 publications

(5 citation statements)

References 34 publications

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“…Other serological tests use crude preparations derived from bloodstream form trypomastigotes grown in vivo (mice or rats) or propagated in vitro [67, 68]. The CFT test for dourine, using whole cell extracts from a T. equiperdum strain, has the advantage that it, to a certain level, also reacts with antibodies against T. evansi ; however, this also entails the risk of false positive reactions [67].…”

Section: Serological Diagnosismentioning

confidence: 99%

“…Among scientist members of the OIE Non-Tsetse Transmitted Animal Trypanosomoses Network [71], the consensus is that the Onderstepoort Veterinary Institute (OVI) strain can be adopted as reference strain for T. equiperdum for the purpose of CFT antigen preparation. It is this strain that is now used by the European Reference Laboratory for Equine Diseases in France, the National Reference Laboratory for Dourine in Germany and the USDA National Veterinary Services Laboratories [67, 68]. New Mongolian T. equiperdum isolates have not been fully typed; whether they may serve as reference strains for the Asian region remains to be investigated [16].…”

Section: Serological Diagnosismentioning

confidence: 99%

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Equine trypanosomosis: enigmas and diagnostic challenges

et al. 2019

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Equine trypanosomosis is a complex of infectious diseases called dourine, nagana and surra. It is caused by several species of the genus Trypanosoma that are transmitted cyclically by tsetse flies, mechanically by other haematophagous flies, or sexually. Trypanosoma congolense (subgenus Nannomonas ) and T. vivax (subgenus Dutonella ) are genetically and morphologically distinct from T. brucei , T. equiperdum and T. evansi (subgenus Trypanozoon ). It remains controversial whether the three latter taxa should be considered distinct species. Recent outbreaks of surra and dourine in Europe illustrate the risk and consequences of importation of equine trypanosomosis with infected animals into non-endemic countries. Knowledge on the epidemiological situation is fragmentary since many endemic countries do not report the diseases to the World Organisation for Animal Health, OIE. Other major obstacles to the control of equine trypanosomosis are the lack of vaccines, the inability of drugs to cure the neurological stage of the disease, the inconsistent case definition and the limitations of current diagnostics. Especially in view of the ever-increasing movement of horses around the globe, there is not only the obvious need for reliable curative and prophylactic drugs but also for accurate diagnostic tests and algorithms. Unfortunately, clinical signs are not pathognomonic, parasitological tests are not sufficiently sensitive, serological tests miss sensitivity or specificity, and molecular tests cannot distinguish the taxa within the Trypanozoon subgenus. To address the limitations of the current diagnostics for equine trypanosomosis, we recommend studies into improved molecular and serological tests with the highest possible sensitivity and specificity. We realise that this is an ambitious goal, but it is dictated by needs at the point of care. However, depending on available treatment options, it may not always be necessary to identify which trypanosome taxon is responsible for a given infection.

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Section: Serological Diagnosismentioning

confidence: 99%

Section: Serological Diagnosismentioning

confidence: 99%

Equine trypanosomosis: enigmas and diagnostic challenges

et al. 2019

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“…The peak trypanosome concentration with 1% FCeM supplementation (Day 4, 3.95 ± 0.43 × 10 6 cells/mL) was significantly higher in comparison to the peak concentrations with the supplementation of other concentrations of FCeM on day 5 (0% FCeM, 1.54 ± 0.32 × 10 6 ; 0.01% FCeM, 1.57 ± 0.34 × 10 6 ; 0.1%, 2.19 ± 0.23 × 10 6 ; and 10%, 2.23 ± 0.93 × 10 6 cells/mL) (Fig. 1), and was almost the same as the cell concentration reported in a previous study (Bassarak et al., 2016). These results suggested that 1% FCeM supplementation in HMI-9 media was also the optimum media for the liquid culture of T. equiperdum .…”

Section: Discussionmentioning

confidence: 86%

“…Bassarak et al. succeeded in establishing an efficient in vitro T. equiperdum culture system in a method that focused on the trypanosome metabolism by supplementation with a trace additive (Bassarak et al., 2016). Recently, a new T. equiperdum strain, T. equiperdum IVM-t1, was established using a soft agarose culture system that was described in our previous study (Suganuma et al., 2016).…”

Section: Discussionmentioning

confidence: 99%

The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum

Suganuma

Yamasaki

Molefe

et al. 2017

International Journal for Parasitology: Drugs and Drug Resistan

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Dourine is caused by Trypanosoma equiperdum via coitus with an infected horse. Although dourine is distributed in Equidae worldwide and is listed as an internationally important animal disease by the World Organization for Animal Health (OIE), no effective treatment strategies have been established. In addition, there are no reports on drug discovery, because no drug screening system exists for this parasite. A new T. equiperdum strain was recently isolated from the genital organ of a stallion that showed typical symptoms of dourine. In the present study, we adapted T. equiperdum IVM-t1 from soft agarose media to HMI-9 liquid media to develop a drug screening assay for T. equiperdum. An intracellular ATP-based luciferase assay using CellTiter-Glo reagent and an intracellular dehydrogenase activity-based colorimetric assay using WTS-8 tetrazolium salt (CCK-8 reagent) were used in order to examine the trypanocidal effects of each compound. In addition, the IC50 values of 4 reference trypanocidal compounds (pentamidine, diminazene, suramin and melarsomine) were evaluated and compared using established assays. The IC50 values of these reference compounds corresponded well to previous studies involving other strains of T. equiperdum. The luciferase assay would be suitable for the mass screening of chemical libraries against T. equiperdum because it allows for the simple and rapid-evaluation of the trypanocidal activities of test compounds, while a simple, inexpensive colorimetric assay will be applicable in developing countries for the evaluation of the drug sensitivity of epidemic trypanosome strains.

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“…Significant improvements in Dourine sero-diagnosis will require the development of more specific T. equiperdum-subunit antigens. A recent publication reported the successful in vitro cultivation of T. equiperdum OVI parasites that can be used in complement fixation tests (Bassarak et al, 2016). This might be of great help in obtaining a specific diagnosis for T. equiperdum, since the OVI strain is one of the few genuine T. equiperdum strains generally available in reference laboratories (Claes et al, 2003b).…”

Section: Serology: Indirect Diagnosismentioning

confidence: 99%

Trypanozoma equiperdum bij het paard – een onbekende bedreiging?

Ahmed¹,

Hagos²,

Merga³

et al. 2018

Vlaams Diergeneeskundig Tijdschrift

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Dourine is a contagious disease caused by Trypanosoma equiperdum that is transmitted directly from animal to animal during coitus. Dourine is known as an important disease in many countries, and it threatens equidae worldwide. It is reported to be widespread in South America, Eastern Europe, Russia, Mongolia, Namibia and Ethiopia. The disease can be carried to various parts of the world through the transportation of infected animals and semen. Since knowledge of the prepatent infectiousness of a recently infected animal is lacking, introduction of the disease is in principle an ever-present threat. Definitive diagnosis depends on the identification of the parasite by means of direct microscopy. This is rarely possible in practice and therefore, diagnosis in the field is based on the observation of typical clinical signs, together with serological tests. This paper is an endeavour to review briefly and compile information on the appearance and importance of Dourine in terms of its epidemiological and clinical features, as well as on its diagnosis, treatment and prognosis.

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In vitro production of Trypanosoma equiperdum antigen and its evaluation for use in serodiagnosis of dourine

Cited by 4 publications

References 34 publications

Equine trypanosomosis: enigmas and diagnostic challenges

Equine trypanosomosis: enigmas and diagnostic challenges

The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum

Trypanozoma equiperdum bij het paard – een onbekende bedreiging?

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