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. 2020 Nov 26;10:20657. doi: 10.1038/s41598-020-77687-7

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Immunolocalization of F. hepatica KT in infective NEJs in the first 48 h of infection. (A–D) show a plane within the interior of the NEJ, whereas (E–H) show a plane at the surface of the same individual NEJs. Following excystment, NEJs were maintained in culture and sample parasites taken at 3 h post-excystment (B,F), 24 h post-excystment (C,G) and 48 h post-excystment (D,H). Parasites were probed with rabbit pre-immune serum (A,E) or anti-FhKT1 antibodies followed by FITC-labelled secondary antibodies. The presence of FhKT within F. hepatica-specific structures are indicated by arrows; parenchymal cell bodies, white arrows; NEJ digestive tract, grey arrows; network of channels below surface tegument, black arrows. All specimens were counter-stained with phalloidin-tetramethylrhodamine isothiocyanate (TRITC) to stain muscle tissue (red fluorescence) and provide structure. OS oral sucker, VS ventral sucker. Scale bars 20 μM.