Successful storage of Trichomonas gallinae on Whatman FTA cards following culture

Abstract

Logistical constraints concerning parasite sample storage can hinder progress with the discovery of genetic variation on a global scale. New storage methods are being developed to address this, but require testing in order to understand how widely applicable these methods are. Whatman FTA cards have been tested previously under laboratory conditions for storing low-concentration Trichomonas gallinae isolates with the conclusion that they are not suitable, but have not been tested under field conditions. Here, we conducted a field-test, comparing FTA cards with storage in ethanol for T. gallinae samples collected and cultured from wild Columbiformes in Africa using standard field methods, before transportation to the UK. After 6 months storage, both methods resulted in an overall prevalence of 100% following PCR amplification (n = 59), suggesting that FTA cards are suitable for estimation of T. gallinae prevalence. However, samples stored in ethanol produced more, and longer, sequences than those stored on FTA cards. These data suggest storage in ethanol is preferable for the acquisition of high quality genetic strain data, but that FTA cards can be used successfully to ascertain infection prevalence and identify parasite strains under field conditions.

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“…Trichomonas gallinae samples collected in Senegal in 2014 were immediately isolated from the media after incubation, as described below. Each T. gallinae sample collected in Senegal in 2015 was split, and half stored with equal amounts of ethanol in an Eppendorf and half on Whatman FTA Classic cards (GE Healthcare Life Sciences), as part of a separate project and as described in Thomas et al (2022), before being imported to the UK where parasites were isolated from the media.…”