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Fig. 19. Light micrographs through gill glands.A in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 19. Light micrographs through gill glands.A, male Attonitus bounites, SL 43.0 mm (USNM 349701), longitudinal section through gill gland showing the ventral epithelium (arrows) that results in the formation of gill gland chambers (c); arrowhead indicates an unmodified gill filament; bar, 200 µm. B, same specimen showing tall columnar cells (arrows) in between reduced secondary lamellae; s, secretory material; bar, 50 µm. C, male A. irisae, SL 46.1 mm (USNM 349698), longitudinal section through immature gill gland showing the epithelium (arrows) covering the gland chambers (c); bar, 200 µm.

Fig. 17 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 17. Scanning electron micrograph of spermatozoon in testis of male Bryconadenos tanaothoros (MCP 30333); arrow indicates point of exit of the flagellum from the cytoplasmic collar; bar, 1 µm.

Fig. 18 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 18. Transmission electron micrographs through spermatozoa from mature male Bryconadenos tanaothoros, SL 50.0 mm (USNM 352061). A-D, successively more posterior transverse sections; E-F, longitudinal sections. Symbols: n, nucleus; m, mitochondrion; p, proximal centriole; d, distal centriole; f, flagellum, arrows, accessory microtubules; arrowheads, cytoplasmic canal of collar. Bars: A-C, 0.2 µm; D, 0.1 µm; E-F, 0.5 µm.

Fig. 14 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 14. Knodus sp., MZUSP 38004. Inseminating species. Caudal-fin squamation. Male, SL 34.0 mm, lateral view left side, portraying scales moderately extended out onto caudalfin rays. Note: lateral line incomplete and not present in region shown. Brazil: Minas Gerais, riacho near Fazenda Poderosa, tributary to ribeirão Jaboticatubas, tributary to rio das Velhas, Campo Alegre.

Fig. 16 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 16. Light micrographs through mature ovaries of the three species of Attonitus; arrows, spermatozoa; bar, 10 µm. A, Attonitus ephimeros, SL 33.1 mm (USNM 349696). B, A. irisae, SL 36.6 mm (USNM 349704). C, A. bounites, SL 43.0 mm (USNM 349701).

Fig. 12. Bryconamericus iheringii, USNM 285884 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 12. Bryconamericus iheringii, USNM 285884. Non-inseminating species. Caudal-fin squamation. Male, SL 53.2 mm, lateral view left side, portraying scales confined to caudal-fin base. Brazil: Rio Grande do Sul, Arroio Garupá, tributary to rio Quaraí, tributary of rio Uruguay. Road: Uruguaiana-Quaraí, locality between Uruguaiana and Quaraí.

Fig. 7 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 7. Bryconadenos tanaothoros, new species, USNM 352061, paratype, SL 36.6 mm, adult male. Lateral view of left side of anterior region of anal fin illustrating glandular organ. Brazil: Mato Grosso: tributary to rio Suiazinho, near Ribeirão Cascalheira.

Fig. 1 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 1. Bryconadenos tanaothoros, two live adult male specimens in aquaria. Originally collected in Brazil: Mato Grosso; tributary to rio Suiazinho near Ribeirão Cascalheira. The reflective green shown especially by the lower specimen of the two sexually active males displays only weakly this color of the body sides. The position of the anal-fin gland can be seen as a pale gray-white oval area just posterior to the proximal end of the white pigment along the anterior border of the anal fin, especially in the upper male in the picture. Both males display the lateral-line pores and tubes outlined in black as occurs in live specimens.

Fig. 5 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 5. Bryconadenos tanaothoros, new species, paratype. USNM 352061, SL 34.7 mm, adult male. Lateral view of left side of anterior region of anal fin showing fusion of pterygiophores and respective medial radials of anterior five branched rays. Note that the pterygiophores lack anterior and posterior median blades for the origin of the erectors and depressors anales muscles. Brazil: Mato Grosso; tributary to rio Suiazinho near Ribeirão Cascalheira.

Fig. 4 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 4. Bryconadenos tanaothoros, new species, MZUSP 85852, SL 41.3 mm, adult male, holotype. Brazil: Mato Grosso, Serra do Roncador, tributary of rio Suiá-Missu, rio Xingu drainage.

Fig. 9 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 9. Bryconadenos tanaothoros, new species, USNM 352061, paratype, SL 33.7 mm, adult male. Jaws and dentition lateral view, left side. Brazil: Mato Grosso: tributary to rio Suiazinho, near Ribeirão Cascalheira.

Fig. 3 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 3. Bryconadenos tanaothoros, new species, MCP 30516, SL 32.0 mm, adult female. Brazil: Mato Grosso, córrego Tatu, on the road MT-423, GPS coordinates 11°28'36"S 54°58'47"W. Adult female freshly fixed in formalin. The dark chromatophores of this specimen associated with the lateral line are displayed almost evenly throughout the lateral line's length. The anal fin shows no evidence of a gland.

Fig. 2 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 2. Bryconadenos tanaothoros, new species, MCP 29476, SL 43.5 mm, adult male. Brazil: Mato Grosso, rio Ferro on the road between Novo Mato Grosso and Nova Ubiratã, about 25 km SW of Novo Mato Grosso, upper rio Xingu basin, GPS coordinates 13°3'32"S 55°2'12"W. Sexually active male specimen freshly fixed in formalin. The dark chromatophores of this specimen associated with the lateral line are contracted and barely visible anteriorly and do not show at all posteriorly. The position of the anal-fin gland can be seen as a yellowish brown oval area just posterior to the proximal end of the white pigment along the anterior border of the anal fin.

Fig. 10 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 10. Pelvic fin length as a function of SL by sex for males = S and females = z of Bryconadenos tanaothoros. For explanation see text under sexual dimorphism.

Fig. 8 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 8. Bryconadenos tanaothoros, new species, USNM 352061, paratype, SL 34.4 mm, adult female. Lateral view of left side of anterior region of anal fin illustrating absence of glandular organ. Brazil: Mato Grosso: tributary to rio Suiazinho, near Ribeirão Cascalheira.

Fig. 6 in Putative relationships among inseminating and externally fertilizing characids, with a description of a new genus and species of Brazilian inseminating fish bearing an anal-fin gland in males (Characiformes: Characidae)

Fig. 6. Attonitus irisae, Vari & Ortega USNM 349698, SL 45.5 mm, adult male. Lateral view of left side of entire anal fin showing fusion of pterygiophores and respective medial radials of anterior two branched rays. Note that the pterygiophores bear anterior and posterior median blades for the origin of the large erectors and depressors anales muscles. Peru: Huanuco; Pachitea, río Huambo at mouth into río Pachitea.

Fig. 4 in Resolution of the identity of three species of Diplostomum (Digenea: Diplostomidae) parasitising freshwater fishes in South Africa, combining molecular and morphological evidence

Fig. 4. Metacercariae of Diplostomum spp. from eye lenses of different fish hosts; (a) Diplostomum sp. from Tilipia sparrmanii, live, ventral view (b) Diplostomum sp. from Tilipia sparrmanii, fixed, ventral view (c) Diplostomum sp. from Tilipia sparrmanii, live, sunken pseudosuckers (arrowhead) (hologenophore, GenBank MN813526, MN813534 and MN808616) (d) Diplostomum sp. 14 sensu Locke et al. (2015) from Synodontis zambezensis, live, ventral view (hologenophore, GenBank MN813541) (e) Diplostomum sp. 14 sensu Locke et al. (2015) from Oreochromis mossambicus, fixed, ventral view, small excretory granules (arrowhead) (hologenophore, GenBank MN813531, MN813539 and MN808621) (f) Diplostomum sp. 14 sensu Locke et al. (2015) from Synodontis zambezensis, fixed, ventral view, large excretory granules (arrowhead) (hologenophore, GenBank MN813541) (g) Diplostomum sp. 16 sensu Locke et al. (2015) from Pseudocrenilabrus philander, fixed, ventral view, everted pseudosuckers (arrowhead) (hologenophore, GenBank MN813532, MN813547 and MN808627) (h) Diplostomum sp. 16 sensu Locke et al. (2015) from Pseudocrenilabrus philander, fixed, ventral view, inverted pseudosuckers (arrowhead) (hologenophore, GenBank MN813533, MN813548 and MN808628) (i) Diplostomum sp. 16 sensu Locke et al. (2015) from Pseudocrenilabrus philander, live metacercariae inside of fish lens. Scale bars: a–h = 100 μm; i = 700 μm.

Fig. 3 in Resolution of the identity of three species of Diplostomum (Digenea: Diplostomidae) parasitising freshwater fishes in South Africa, combining molecular and morphological evidence

Fig. 3. Bayesian inference (BI) and maximum likelihood (ML) phylogram reconstructed using cox1 sequences for species of Diplostomum. Nodal support from BI and ML analyses indicated as BI/ML; only values> 0.90 (BI) and> 70 (ML) are displayed. Scale-bar indicates the expected number of substitution per site. Sequences generated in this study are in bold and indicated by blue rectangles. Codes with isolate information for newly generated sequences are provided in Table 3. Sequences derived from Africa are highlighted in blue, from Asia in purple, from Europe in orange, from North America in green (according to the map) and sequences reported from more than one continent are highlighted in black. Black arrows on the map demonstrate distribution of Diplostomum spathaceum and 'D. mergi Lineage 2' in both, Asia and Europe, and Diplostomum sp. 14 and Diplostomum sp. 16 in both, Africa and Asia. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 2 in Resolution of the identity of three species of Diplostomum (Digenea: Diplostomidae) parasitising freshwater fishes in South Africa, combining molecular and morphological evidence

Fig. 2. Bayesian inference (BI) and maximum likelihood (ML) phylograms reconstructed using (a) partial 28S rDNA sequences (b) ITS1-5.8S-ITS2 sequences for species of Diplostomum. Nodal support from BI and ML analyses indicated as BI/ML; only values> 0.90 (BI) and> 70 (ML) are displayed. Scale-bar indicates the expected number of substitution per site. Sequences generated in this study are in bold and indicated by blue rectangles. Codes with isolate information for newly generated sequences are provided in Table 3. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 1 in Resolution of the identity of three species of Diplostomum (Digenea: Diplostomidae) parasitising freshwater fishes in South Africa, combining molecular and morphological evidence

Fig. 1. Map illustrating the sampling localities on (a) River Riet in Mokala National Park (b) River Phongolo (Site 1, Site 2 and Nyamithi Lake) and the River Usuthu (Shokwe Pan) in Ndumo Game Reserve and (c) River Mooi (Boskop Dam) in Boskop Dam Nature Reserve, South Africa. The illustration was compiled in ArcGIS 10.6 (Available from https://support.esri.com/en/downloads).