Fig 1.
In vitro motility assay with adult Trichuris muris.
The motility score is given as arithmetic means (± SD) of three individual studies where worms (n = 30) were incubated in RPMI medium with (+) or without (-) glucose (glu) and/or amino acids (aa) for 17 (Fig 1a) or 41 (Fig 1b) hours. ***: p < 0.001. No variation in motility was observed for worms incubated in +aa, +glu.
Fig 2.
In vitro Alamar Blue assay with adult Trichuris muris.
Emission of the reduced dye resorufin is given as arithmetic means (± SD) of three individual studies where worms (n = 4) were incubated in RPMI medium with (+) or without (-) glucose (glu) and/or amino acids (aa) for 17 (Fig 2a) or 41 (Fig 2b) hours.
Table 1.
Estimated effects of glucose, amino acids and time on the viability of Trichuris muris using a linear mixed model (with replicate as a random effect).
Fig 3.
Confocal image showing the ventral side of the thin anterior part of an adult Trichuris muris.
The bacillary band was labelled with Alexa Fluor 633 (100 μg/mL) for 30 minutes and can be seen as red circles or dots. Scale bar: 100 μm.
Fig 4.
Confocal image of the ventral side of an adult Trichuris muris showing the pores of the bacillary band (Fig 4a) and accumulated 6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-6-Deoxyglucose (6-NBDG) (Fig 4b) in the pores of the bacillary band (Fig 4c) after incubation in agarose (1%) with 6-NBDG (200 μM) for approximately 5 minutes.
Fluorescence from 6-NBDG and Alexa Fluor 633 is shown in green and red respectively. Scale bar: 30 μm.
Fig 5.
Transmission images (Fig 5a and d) and confocal section through the thin anterior part (Fig 5b and c) and oral cavity (Fig 5e and f) of an adult Trichuris muris not exposed to 6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-6-Deoxyglucose (6-NBDG) is shown for emission wavelengths between 500–600 (6-NBDG, Fig 5c and f) and 642–700 nm (Alexa Fluor 633, Fig 5b and e).
Fig 6.
Confocal section through the thin anterior part of an adult Trichuris muris after incubation in agarose (1%) with 6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-6-Deoxyglucose (6-NBDG) (200 μM) for 6 (Fig 6a), 15 (Fig 6b) and 30 (Fig 6c) minutes.
The glucose analogue 6-NBDG was absorbed by the bacillary band and accumulated within the stichocytes in which the mean grey value increased from 8.7 (Fig 6a) to 12.8 (Fig 6b) and 13.5 (Fig 6c). Scale bar: 30 μm.
Fig 7.
Confocal section through the thin anterior part of an adult Trichuris muris showing the accumulation of 6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-6-Deoxyglucose (6-NBDG) within the bacillary band, the stichocytes and the connecting membranes after incubation in agarose (1%) with 6-NBDG (200 μM) for approximately 30 minutes.
Scale bar: 50 μm.
Fig 8.
Transmission image (Fig 8a and 8c) and confocal section (Fig 8b and 8d) through an adult Trichuris muris at the junction between the thin anterior and the thick posterior part of the worm after incubation in agarose (1%) with 6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-6-Deoxyglucose (6-NBDG) (200 μM) for approximately 40 minutes.
Note the eggs are located ventrally near the vulva, which is itself located at the border between the thin anterior and the thick posterior part of the worm (Fig 8a and 8b). The bacillary band was not visible in this area and the stichocytes and the connective membranes were devoid of 6-NBDG. Scale bar: 50 μm. High magnification of the stichocytes and the connective membranes devoid of 6-NBDG (Fig 8c and 8d). Scale bar: 30 μm.
Fig 9.
Confocal section through the thin anterior part of an adult Trichuris muris with sealed oral cavity after incubation in agarose (1%) with 6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-6-Deoxyglucose (6-NBDG) (200 μM) for 29 (Fig 9a), 44 (Fig 9b) and 56 (Fig 9c) minutes.
Despite the sealed oral cavity, 6-NBDG accumulated within the stichocytes with mean grey values increasing from 5.8 to 8.1 and 9.4. Scale bar: 50 μm.
Fig 10.
Confocal section through the thin anterior part of an adult worm of Trichuris muris with sealed oral cavity after incubation in agarose (1%) with 6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-6-Deoxyglucose (6-NBDG) (200 μM) for approximately 40 minutes.
Despite the sealed oral cavity, 6-NBDG accumulated within the stichocytes and the connective membranes. Scale bar: 50 μm.