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Fig. 5 in Redescription of Bicotyle reticulata (Monogenea: Heteraxinidae) from Pampus punctatissimus (Scombriformes: Stromateidae) in the Seto Inland Sea, Japan

Fig. 5. Bayesian inference (BI) tree for the Microcotylinea based on partial 28S rDNA data (840 bp) using two species of Plectanocotylidae as the outgroup. The species newly sequenced in this study is indicated in bold. The corresponding INSD accession numbers are shown. The tree includes results for Bayesian inference and ML with PP/BS branch support values.

Fig. 2 in Redescription of Bicotyle reticulata (Monogenea: Heteraxinidae) from Pampus punctatissimus (Scombriformes: Stromateidae) in the Seto Inland Sea, Japan

Fig. 2. Bicotyle reticulata (Goto, 1894) from Pampus punctatissimus (Temminck and Schlegel, 1845). A, Clamp on right side (closed, ventral view, MPM Coll.-No. 25262); B, clamp on left side (open, apical view, MPM Coll.-No. 25262); C, mouth (MPM Coll.-No. 25262); D, genital atrium (MPM Coll.-No. 25261); E, vaginal pore (MPM Coll.-No. 25261). Scale bars: A, B, 50 µm; C, 600 µm; D, E, 250 µm.

Fig. 1 in Redescription of Bicotyle reticulata (Monogenea: Heteraxinidae) from Pampus punctatissimus (Scombriformes: Stromateidae) in the Seto Inland Sea, Japan

Fig. 1. Bicotyle reticulata (Goto, 1894) from Pampus punctatissimus (Temminck and Schlegel, 1845). Whole body (ventral view, MPM Coll.-No. 25262). Scale bar: 10 mm.

Fig. 4 in Redescription of Bicotyle reticulata (Monogenea: Heteraxinidae) from Pampus punctatissimus (Scombriformes: Stromateidae) in the Seto Inland Sea, Japan

Fig. 4. Digestive tract of Bicotyle reticulata (Goto, 1894) from Pampus punctatissimus (Temminck and Schlegel, 1845) (ventral view, MPM Coll.-No. 25262). Scale bar: 5 mm.

Fig. 3 in Redescription of Bicotyle reticulata (Monogenea: Heteraxinidae) from Pampus punctatissimus (Scombriformes: Stromateidae) in the Seto Inland Sea, Japan

Fig. 3. Bicotyle reticulata (Goto, 1894) from Pampus punctatissimus (Temminck and Schlegel, 1845). Reproductive organs (ventral view, MPM Coll.-No. 25262). Scale bar: 500 µm.

Fig. 4. Chloeia parva chaetae. A–L in Pampus candidus

Fig. 4. Chloeia parva chaetae. A–L: specimen SWIMS-ANN-19-003. (A) bifurcate notochaetae, chaetiger 2, left side; (B) bifurcate notochaetae, chaetiger 4, left side; (C) bifurcate notochaetae, chaetiger 5, right side; (D) harpoon notochaetae, chaetiger 6, left side; (E) harpoon notochaetae, chaetiger 10, left side; (F) harpoon notochaetae, chaetiger 17, left side; (G) harpoon notochaetae, chaetiger 30, left side; (H) spinose notochaetae, chaetiger 5, 17 and 30, respectively; (I) bifurcate neurochaetae, chaetiger 2, left side; (J) bifurcate neurochaetae, chaetiger 10, left side; (K) bifurcate neurochaetae, chaetiger 17, left side; (L) bifurcate neurochaetae, chaetiger 30, left side. Scale bars: A–C, I–L = 50 μm; D–H = 100 μm.

Fig. 3. Chloeia parva. A, J in Pampus candidus

Fig. 3. Chloeia parva. A, J, specimen SWIMS-ANN-19-005; B–I, K, specimen SWIMS-ANN-19-003. (A) living specimen, dorsal view; (B) anterior part, dorsal view; (C) anterior part, ventral view; (D) chaetiger 7–9, ventral view, right side; (E) pygidium, ventral view, showing two finger-shaped pygidial cirri; (F) pygidium, dorsal view, showing anus (black arrow); (G) parapodium of chaetiger 2, right side, posterior view; (H) parapodium of chaetiger 10, right side, posterior view; (I) chaetiger 15–16, dorsal view, showing the pattern of dorsal pigmentation; (J) caruncle, dorsal view; (K) chaetiger 15–16, dorsal view, showing branchiae and the pattern of dorsal pigmentation. Abbreviations: an, anus; apl, anterior pigmented line; bc, branchial cirrus; br, branchia; car, carucle; gl, guard line; ipl, inner-posterior pigmented line; lAn, lateral antenna; lg, longitudinal groove; mAn, median antenna; mdm, mid-dorsal mark; nec, neuropodial cirrus; noc, notopodial cirrus; opl, outer-posterior pigmented line; pa, palp; pAn, palpal antenna; pc, pygidial cirrus. Scale bars: A = 10 mm; I = 5 mm; H = 2 mm; B–G, J, K = 1 mm.

Fig. 2 in Pampus candidus

Fig. 2. Chloeia bimaculata sp. nov. chaetae. A–L: holotype (SWIMS-ANN-19-001). (A) bifurcate notochaetae, chaetiger 2, right side; (B) bifurcate notochaetae, chaetiger 4, left side; (C) harpoon notochaetae, chaetiger 5, right side; (D) harpoon notochaetae, chaetiger 6, left side; (E) harpoon notochaetae, chaetiger 10, left side; (F) harpoon notochaetae, chaetiger 17, left side; (G) harpoon notochaetae, chaetiger 23, left side; (H) spinose notochaetae, chaetiger 5, 17 and 23, respectively; (I) bifurcate neurochaetae, chaetiger 2, right side; (J) bifurcate neurochaetae, chaetiger 10, right side; (K) bifurcate neurochaetae, chaetiger 17, left side; (L) bifurcate neurochaetae, chaetiger 23, left side. Scale bars: A–B, I–L = 50 μm; C–G = 200 μm; H = 100 μm.

Fig. 1 in Pampus candidus

Fig. 1. Chloeia bimaculata sp. nov. A–I, holotype (SWIMS-ANN-19-001); J–L, paratype (SWIMS-ANN-19-002). (A) living specimen, dorsal view; (B) anterior part, dorsal view; (C) anterior part, ventral view, showing two small pigment spots on chaetiger 2; (D) chaetiger 9–11, ventral view, left side; (E) pygidium, ventral view, showing two digitiform pygidial cirri; (F) pygidium, dorsal view, showing anus; (G) parapodium of chaetiger 2, left side, posterior view; (H) parapodium of chaetiger 10, left side, posterior view; (I) chaetiger 6–7, dorsal view, showing dorsal cirri and the pattern of dorsal pigmentation; (J) prostomium, showing antennae and palps; (K) anterior part, ventral view, showing black spots on chaetiger 2; (L) chaetiger 6, dorsal view, showing mid-dorsal pigmentation; (M) chaetiger 7–8, dorsal view, showing dorsal pigmentation pattern. Abbreviations: an, anus; apl, anterior pigmented line; bc, branchial cirrus; br, branchia; car, carucle; gl, guard line; ipl, inner-posterior pigmented line; lAn, lateral antenna; lg, longitudinal groove; mAn, median antenna; mds, mid-dorsal spots; nec, neuropodial cirrus; noc, notopodial cirrus; pa, palp; pAn, palpal antenna; pc, pygidial cirrus; opl, outer-posterior pigmented line; sp, spots. Scale bars: A = 10 mm; B–I, K–M = 1 mm; J = 250 μm.

Fig. 5 in Pampus candidus

Fig. 5. Maximum likelihood (ML) phylogenetic trees based on COI (A), 16S (B), 28S (C) and their concatenated sequences (D). Numbers on the branches represent ML bootstrap values (maximum: 100) based on 1000 replicates. GenBank accession numbers of the COI, 16S and 28S genes used are shown in parentheses. Scale bar corresponds to the estimated mean number of nucleotide substitutions per site.

Fig. 11 in Pampus candidus

Fig. 11. Corrolations between revealing mount behavior with the stimulus estrous female during partner preference and expression of ERα, as well as ERβ in mPOA in FC, FT and MC groups. mPOA: medial preoptic area; ERa = estrogen receptor-α; ERb = estrogen receptor β; 1 = FC; 2 = FT; 3 = MC; FRFMB = revealing mount with the stimulus awake estrous female during final partner preference test; A: Corrolations between ERa mPOA and FRFMB, r = -0.545, P = 0.002; B: Corrolations between ERb mPOA and FRFMB, r = 0.371, P = 0.044.

Fig. 10 in Pampus candidus

Fig. 10. Corrolations between received mount behavior with the stimulus awake intact male during partner preference and expression of ERα, as well as ERβ in mPOA in FC, FT and MC groups. mPOA: medial preoptic area; ERa = estrogen receptor-α; ERb = estrogen receptor β; 1 = FC; 2 = FT; 3 = MC; FRMMB = receiving mount with the stimulus awake intact male during final partner preference test; A: Corrolations between ERa mPOA and FRMMB, r = 0.645, P <0.001; B: Corrolations between ERb mPOA and FRMMB, r = -0.651, P <0.001.

Fig. 9 in Pampus candidus

Fig. 9. Corrolations behaviors between preferences scores of awake animals and ERα, as well as ERβ in mPOA in FC, FT and MC groups. mPOA: medial preoptic area; ERa = estrogen receptor-α; ERb = estrogen receptor β; 1 = FC; 2 = FT; 3 = MC; FPWEF = final preference awake estrous female; FPWIM = final preference awake intact male; A: Corrolations between ERa mPOA and FPWEF, r = -0.625, P <0.001; B: Corrolations between ERa mPOA and FPWIM, r = 0.864, P <0.001; C: Corrolations between ERb mPOA and FPWEF, r = 0.659, P <0.001; D: Corrolations between ERb mPOA and FPWIM, r = -0.744, P <0.001.

Fig. 8 in Pampus candidus

Fig. 8. Correlations behaviors between preferences scores of anesthetize animals and ERα, as well as ERβ in mPOA in FC, FT and MC groups. mPOA: medial preoptic area; ERa = estrogen receptor-α; ERb = estrogen receptor β; 1 = FC; 2 = FT; 3 = MC; FPAEF = final preference anesthetize estrous female; FPAIM = final preference anesthetize intact male; A: Correlations between ERa mPOA and FPAEF, r = -0.610, P <0.001; B: Correlations between ERa mPOA and FPAIM, r = 0.790, P <0.001; C: Correlations between ERb mPOA and FPAEF, r = 0.636, P <0.001; D: Correlations between ERb mPOA and FPAIM, r = -0.678, P <0.001.

Fig. 6 in Pampus candidus

Fig. 6. Mean (± SE) number of ERα-IRs in FC, FT, and MC groups. mPOA: medial preoptic area. BNST: bed nucleus of the stria terminalis. MeA: medial amygdaloid nucleus. VMH: ventromedial nucleus of the hypothalamus. 3V: third ventricle. OT: optic tract. Scale bar = 200 μm. *: P <0.05, **: P <0.01, ***: P <0.001.

Fig. 5 in Pampus candidus

Fig. 5. Frequency of proceptive behaviors shown by experimental females during the female sexual behavior test. FT females exhibited fewer proceptive behaviors than FC females. ***: Significantly different from control group, P <0.001. See text for details.

Fig. 7 in Pampus candidus

Fig. 7. Mean (± SE) number of ERβ-IRs in FC, FT, and MC groups. mPOA: medial preoptic area. BNST: bed nucleus of the stria terminalis. MeA: medial amygdaloid nucleus. VMH: ventromedial nucleus of the hypothalamus. 3V: third ventricle. LV: lateral ventricle. OT: optic tract. Scale bar = 200 μm. *: P <0.05, **: P <0.01, ***: P <0.001.

Fig. 4 in Pampus candidus

Fig. 4. Frequencies of mounts, intromissions, and ejaculations in females receiving sexual behavior tests. Males showed fewer behaviors when paired with FT females and MC males compared to FC females. **: Significantly different from control group, P <0.01. ***: Significantly different from control group, P <0.001. See text for details.

Fig. 3 in Pampus candidus

Fig. 3. Partner preference data (Behavioral Test 2). In Behavioral Test 2, experimental females were exposed to perinatal treatments and stimulus females or males were awake. 3A) There were no differences for the time spent in their own chamber for FC females, FT females and MC males. 3B) FT females and MC males showed a higher preference score than that of FC females in partner preference tests. Preference score is calculated as time spent with stimulus female minus the time spent with stimulus male. 3C) FT females and MC males spent more time with the stimulus awake estrous female than did FC females. 3D) FT females and MC males spent less time with the stimulus awake intact male than did FC females. *: Significantly different from control group, P <0.05; **: Significantly different from control group, P <0.01.

Fig. 2 in Pampus candidus

Fig. 2. Partner preference data (Behavioral Test 1). In Behavioral Test 1, experimental females were exposed to perinatal treatments and stimulus females or males were anesthetized. 2A) FT females and MC males spent more time with the stimulus anesthetized estrous female than the FC females did. 2B) FT females and MC males spent less time with the stimulus anesthetized intact male than did FC females. 2C) There was no difference in time spent in their own chamber for FC females, FT females, and MC males. 2D) FT females and MC males showed a higher preference score than that of FC females in the partner preference tests. Preference score is calculated as time spent with stimulus female minus the time spent with stimulus male. *: Significantly different from control group, P <0.05; **: Significantly different from control group, P <0.01.