Abstract
The control and elimination of malaria caused by Plasmodium vivax both represent a great challenge due to the biological aspects of the species. Gametocytes are the forms responsible for the transmission of the parasite to the vector and the search for new strategies for blocking transmission are essential in a scenario of control and elimination The challenges in this search in regard to P. vivax mainly stem from the lack of a long-term culture and the limitation of studies of gametocytes. This study evaluated the viability and infectivity of P. vivax gametocytes in short-term culture. The samples enriched in gametocytes using Percoll (i), using magnetic-activated cell sorting (MACS®) (ii), and using non-enriched samples (iii) were evaluated. After the procedures, gametocytes were cultured in IMDM medium for up to 48 h. Cultured P. vivax gametocytes were viable and infectious for up to 48 h, however differences in viability and infectivity were observed in the samples after 12 h of culture in relation to 0 h. Percoll-enriched samples were shown to be viable in culture for longer intervals than those purified using MACS®. Gametocyte viability after enrichment procedures and short-term culture may provide new avenues in the development of methods for evaluating P. vivax TB.
Highlights
Plasmodium vivax is the leading malaria-causing species worldwide (World Health Organization, 2020) and some severe cases and mortality are reported in endemic are as in tropical and subtropical regions (Alexandre et al, 2010; Howes et al, 2016; Kotepui et al, 2020)
Experimental Infection in Anopheles aquasalis Using a Membrane Feeding Assay (MFA) Cultured gametocytes (4–14 wells for each time period) were centrifuged (400g/5 min/37°C), the medium was removed and the pellet obtained for samples enriched with magnetic cell sorting (MACS)® or Percoll was resuspended in 400 μl of uninfected fresh O+ blood to a final 40% hematocrit with heat-inactivated human AB serum
The gametocyte counts decreased at the intervals of 24 and 48 h, when compared to 0 h (Figure 1A)
Summary
Plasmodium vivax is the leading malaria-causing species worldwide (World Health Organization, 2020) and some severe cases and mortality are reported in endemic are as in tropical and subtropical regions (Alexandre et al, 2010; Howes et al, 2016; Kotepui et al, 2020). P. vivax has unique biological characteristics, such as rapid gametocytogenesis that facilitates transmission and the development of latent forms, known as hypnozoites, which are responsible for relapses months after treatment (Bermúdez et al, 2018; Dhiman, 2019). These features present a great challenge in the control and elimination strategies of vivax malaria. Since P. vivax cannot be cultivated for long periods, ex vivo studies have provided several possibilities for analysis, their focus is mainly on asexual stages, and few results on viability and infectivity are available regarding gametocytes. This study evaluates the viability and infectivity of gametocytes in a short-term ex vivo P. vivax culture
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