Precision mapping: An innovative tool and way forward to shrink the map, better target interventions, and accelerate toward the elimination of schistosomiasis - Fig 1

<p><b>Comparison of precision and conventional maps of schistosomiasis prevalence in the health districts of Edea (A) and Ndikinimeki (B), Cameroon.</b> The precision maps (A1 and B1) provide more accurate information on the distribution of schistosomiasis within districts and a clear precision on subdistricts or communities requiring preventive chemotherapy. Differences of subdistrict prevalence between 1985 and 2010 mappings further illustrate the limitations and uncertainties of the conventional mapping (A2 versus A3 and B2 versus B3). Produced with Esri ArcGIS Pro 2.0.</p

Map showing the location of the survey sites across Senegal.

<p>The survey sties are numbered in red. GPS coordinates: 1. Richard Toll = 16°27′40.71″N, 15°41′15.29″W, 2. Nder = 16°15′33.50″N, 15°53′2.13″W, 3. Linguiere = 15°23′34.33″N, 15°6′55.87″W, 4. Barkedji =  =  15°16′38.46″N, 14°51′55.48″W, 5. Tambacounda = 13°46′8.00″N, 13°40′2.00″W, 6. Kolda = 12°53′58.27″N, 14°56′39.37″W. The regions of Senegal are also shown on the map.</p

Experimental laboratory animal infections.

*<p>Hamsters were used as <i>S. haematobium</i> does not develop well in mice.</p>**<p>Genetic profiles and number of hybrid miracidia resulting from the heterospecific crosses. In total 96 miracidia from each animal cross were genetically analysed.</p><p><i>S. c</i> = <i>S. curassoni</i>, <i>S. b</i> = <i>S. bovis</i>, <i>S. h</i> = <i>S. haematobium</i>, M = male and F = female.</p

Western blot analysis demonstrating antibody specificity and activation and inhibition of <i>S. mamsoni</i> PKA-C.

<p>(A) Adult <i>S. mansoni</i> protein extracts (from one worm pair) were processed for western blotting with anti-phospho-PKA-C antibodies as described in Materials and Methods; the immunizing peptide and lambda phosphatase were employed to confirm that the antibody reacted with phosphorylated PKA-C and not the non-phosphorylated form. (B) Live adult <i>S. mansoni</i> were incubated in 50 µM or 100 µM forskolin, 25 µM or 50 µM KT5720, vehicle (DMSO), or DMEM alone (Control) for 1 h at 28°C and worm protein extracts processed for western blotting with anti-phospho-PKA-C antibodies. Anti-actin antibodies were used to assess any differences in protein loading between samples. The results shown in each panel are representative of data from two independent experiments.</p

Activated PKA associates with the nervous system of adult male <i>S. mansoni</i>.

<p>Schistosomes were fixed and stained with anti-phospho-PKA-C antibodies (green) and rhodamine phalloidin (red) to localize activated PKA and actin, respectively, as detailed in Materials and Methods. Images show z-axis projections in maximum pixel brightness mode with further z, x and y, z projections in D. Intense PKA activation is particularly associated with: (A, anterior view) ventral nerve cords (VNC), connecting cerebral commissures (CC), and anterior ganglia (AG); (A, B) complex nerve plexus (PL) associated with the oral sucker (OS) and extending nerve processes (NP); (C, enlarged from boxed region shown in A) and (D) nerve endings (NE) emanating from the PL evident over the body surface including on the tegument and oral sucker; (E, F) peripheral nervous system including PL extending from the lateral nerve cord (LNC), main nerve cord (MNC), complex PL extending to the underlying musculature (red), surface proximal NEs extending from the PL, nerve fibres (NF); peripheral ganglia (PG) and cell bodies (CB) which connect with the MNC; (F) the ganglion shown was close to the surface of the gut. Additionally, diffuse PKA activation is associated with the (A, G) testicular lobes (TL) of the testes (TT), which are surrounded by the testicular capsules stained red. Bar: A = 100 µm; B–D, F, G = 20 µm; E = 40 µm.</p

<i>In situ</i> distribution of phosphorylated (activated) PKA (green) in adult male <i>S. mansoni</i> revealed using anti-phospho-PKA-C antibodies.

<p>Schistosomes were fixed and stained as detailed in Materials and Methods. Images show z-axis projections in maximum pixel brightness mode except (A) which shows selected serial optical z-sections; images I, J, L, N, and P are overlays of activated PKA with F-actin stained by rhodamine phalloidin (red). (A–C) Negative control worms incubated without primary antibodies but with Alexa Fluor 488 secondary antibodies and rhodamine phalloidin. In male worms, prominent PKA activation is particularly associated with: (D, anterior dorsal surface scan) the tegument (TE); (E, deeper body scan) gynaecophoric canal muscles (GCM) of the gynaecophoric canal (GC), and seminal vesicle (SV); (F, anterior ventral surface scan) ventral sucker (VS), oral sucker (OS) and oesophagus (OE); (G–J) tubercles (TU) of the tegument, areas proximal to the centres of the tubercle spines (SP) and at surface-proximal nerve endings (NE) are also evident in (P); (K, L, N–P) tubular structures (TS) of unknown function close to the SV (K, L) also in the anterior of the worm (N–P, with boxed region shown in O, enlarged for clarity in P); (M) posterior region displaying activated PKA associated with the collecting duct (CD). Bar: B, C = 300 µm; D–F, M = 100 µm; G–L, P = 20 µm; N,O = 50 µm.</p

Activation of PKA in the nervous system of adult male (M) and female (F) <i>S. mansoni</i> upon pair separation.

<p>Schistosomes were fixed (A–C) 15 min, (D–F) 30 min, or (G–I) 60 min after separation and were stained with anti-phospho-PKA-C antibodies (green) to localize activated PKA as detailed in Materials and Methods; schistosomes that remained in copula (pair; C,F,I) for these durations were also processed for comparison with separated males (A,D,G) or females (B,E,H). Images show z-axis projections in maximum pixel brightness mode. The arrows highlight regions of PKA activation within the nervous system; the putative surface-proximal nerve endings (NE) are also indicated.</p

Immunolocalization of activated PKA in forskolin-treated adult male (M) and female (F) <i>S. mansoni in copula</i>.

<p>Worm couples were incubated in 50 µM forskolin for 1 h and fixed and stained with anti-phospho-PKA-C antibodies (green) and rhodamine phalloidin (red) to localize activated PKA, and actin, respectively, as detailed in Materials and Methods. Intense PKA activation is seen in the tegument (TE) musculature and the nervous system including the longitudinal nerve cords (LNC) and nerve endings (NE) covering the TE and oral sucker (OS). The uncharacterized tubular structures (TS) previously show in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0001988#pntd-0001988-g002" target="_blank">Figure 2</a> also display activated PKA. Image shows a z-axis projection in maximum pixel brightness mode. Bar = 100 µm.</p

The origin of microRNAs of <i>Schistosoma mansoni</i>.

<p>A) MicroRNAs characterized from small RNA libraries and their evolutionary origin based on homology searches. B) Example of a newly discovered microRNA in <i>S. mansoni</i> that is also conserved in <i>S. japonicum</i>. Conserved nucleotides between the two species are shown with a black background. The number of uniquely mapped reads for each sequence in <i>S. mansoni</i> is shown in the right, for both SOLiD and MiSeq datasets.</p

Transmission site in Mbane, Senegal.

<p>Hybrid parasites were recovered from snails (<i>Bulinus truncatus</i>) collected at this habitat.</p