Morphological and molecular descriptors of the developmental cycle of Babesia divergens parasites in human erythrocytes

Show simple item record Rossouw, Ingrid Maritz-Olivier, Christine Niemand, Jandeli Van Biljon, Riette Smit, Annel Olivier, Nicholas Abraham Birkholtz, Lyn-Marie
dc.contributor.editor Small, Pamela L.C. 2015-09-04T11:53:59Z 2015-09-04T11:53:59Z 2015-05-08
dc.description S1 Text. Explanation of the development of a fluorescent cell biological evaluation technique to detect intra-erythrocytic B. divergens parasites. en_ZA
dc.description S1 Fig. Flow cytometric analysis of intra-erythrocytic B. divergens parasites. Uninfected erythrocytes were analysed in parallel to B. divergens infected erythrocytes, either fixed (0.025% glutaraldehyde for 45 min) or unfixed. Cells were subsequently stained with either 1:100 and 1:1000 SYBR Green I (30 min, dark, room temperature). (A) Dotblot analysis and (B) histograms of 1) uninfected, unfixed, stained erythrocytes; 2–5) B. divergens infected erythrocytes either unfixed (2 & 3) or fixed (4 & 5). In panels 2 and 4 cells were stained with 1:100 SYBR Green I and in panels 3 and 5 with 1:1000 SYBR Green I. Erythrocytes infected with P. falciparum parasites were comparatively analysed in panel 6 (glutaraldehyde fixed and stained with 1:1000 SYBR Green I. (C) Effect of SYBR Green I concentrations on parasitemia determined from both unfixed and fixed B. divergens infected erythrocytes. Results are the mean of three independent experiments each performed in triplicate (± S.E.). Significance is indicated at P<0.001 ( ) (unpaired Student-t test). (D) Linear correlation analysis (R2 value of 0.98) of B. divergens parasitemia detection between light microscopy and flow cytometry. Data are the mean of three independent experiments each performed in triplicate (± S.E.). Confidence levels (95%) indicated by dashed lines. en_ZA
dc.description S1 Table. Differentially affected transcripts identified in the initiate culture before culture adaptation. en_ZA
dc.description.abstract Human babesiosis, especially caused by the cattle derived Babesia divergens parasite, is on the increase, resulting in renewed attentiveness to this potentially life threatening emerging zoonotic disease. The molecular mechanisms underlying the pathophysiology and intraerythrocytic development of these parasites are poorly understood. This impedes concerted efforts aimed at the discovery of novel anti-babesiacidal agents. By applying sensitive cell biological and molecular functional genomics tools, we describe the intra-erythrocytic development cycle of B. divergens parasites from immature, mono-nucleated ring forms to bi-nucleated paired piriforms and ultimately multi-nucleated tetrads that characterizes zoonotic Babesia spp. This is further correlated for the first time to nuclear content increases during intra-erythrocytic development progression, providing insight into the part of the life cycle that occurs during human infection. High-content temporal evaluation elucidated the contribution of the different stages to life cycle progression. Moreover, molecular descriptors indicate that B. divergens parasites employ physiological adaptation to in vitro cultivation. Additionally, differential expression is observed as the parasite equilibrates its developmental stages during its life cycle. Together, this information provides the first temporal evaluation of the functional transcriptome of B. divergens parasites, information that could be useful in identifying biological processes essential to parasite survival for future antibabesiacidal discoveries. en_ZA
dc.description.librarian am2015 en_ZA
dc.description.sponsorship A South African National Research Foundation ( za) (FA2007050300003 & UID 84627) and Medical Research Council ( grants to LMB as well as the South African National Research Foundation Technology and Human Resources for Industry Programme (THRIP, grant number TP2010072300035) awarded to CMO. IR was supported by a grant holder’s bursary from the National Research Foundation (grant number 73659). en_ZA
dc.description.uri en_ZA
dc.identifier.citation Rossouw I, Maritz-Olivier C, Niemand J, Van Biljon R, Smit A, Olivier NA & Birkholtz, L-M (2015) Morphological and Molecular Descriptors of the Developmental Cycle of Babesia divergens Parasites in Human Erythrocytes. PLoS Neglected Tropical Diseases 9(5): e0003711. DOI:10.1371/journal.pntd.0003711 en_ZA
dc.identifier.issn 1932-6203
dc.identifier.other 10.1371/journal.pntd.0003711
dc.language.iso en en_ZA
dc.publisher Public Library of Science en_ZA
dc.rights © 2015 Rossouw et al. This is an open access article distributed under the terms of the Creative Commons Attribution License. en_ZA
dc.subject Babesia en_ZA
dc.subject Divergens en_ZA
dc.subject Parasites en_ZA
dc.subject Intra-erythrocytic en_ZA
dc.subject Human babesiosis en_ZA
dc.subject Zoonotic disease en_ZA
dc.title Morphological and molecular descriptors of the developmental cycle of Babesia divergens parasites in human erythrocytes en_ZA
dc.type Article en_ZA

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