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Figure 3 in Revision of the genus Chordodes (Gordiida: Nematomorpha) from Africa IV. Ultrastructural redescription of Chordodes congolensis Sciacchitano, 1933, Chordodes ferox Camerano, 1897, Chordodes madagascariensis (Camerano, 1893), Chordodes mobensis Sciacchitano, 1958 and reinterpretation of Chordodes maculatus Sciacchitano, 1958 and Chordodes kakandensis Sciacchitano, 1958

Figure 3. Holotype of Chordodes madagascariensis. (A) Ventral view of the male posterior end, showing the cloacal opening; (B) detail of the male posterior end with the cloacal opening surrounded by circumcloacal bristles and two rows of bristles (bristlefields); (C) posterior end of the female; (D) cuticle of the lateral side of the body with simple, tubercle (arrow) and thorn areoles (double arrow); (E) cluster of crowned areoles on the male. Scale bars: 100 µm (A), 50 µm (B), 200 µm (C), 10 µm (D), 20 µm (E).

Figure 1 in Revision of the genus Chordodes (Gordiida: Nematomorpha) from Africa IV. Ultrastructural redescription of Chordodes congolensis Sciacchitano, 1933, Chordodes ferox Camerano, 1897, Chordodes madagascariensis (Camerano, 1893), Chordodes mobensis Sciacchitano, 1958 and reinterpretation of Chordodes maculatus Sciacchitano, 1958 and Chordodes kakandensis Sciacchitano, 1958

Figure 1. Holotype of Chordodes congolensis. (A) Overview of cuticle showing: simple (1), bulging (2), tubercle (3) and cluster of crowned (4) and circumcluster areoles (5); (B) detail of simple, bulging and tubercle areoles; (C) cluster of two crowned areoles with surrounding circumcluster areoles, arrow indicates thorn areole; (D) crowned areoles with long filaments along the ventral midline; (E) cuticle of female specimen (AMT 1401). Scale bars: 50 µm (A, D), 10 µm (B, E), 20 µm (C).

Figure 6 in Revision of the genus Chordodes (Gordiida: Nematomorpha) from Africa IV. Ultrastructural redescription of Chordodes congolensis Sciacchitano, 1933, Chordodes ferox Camerano, 1897, Chordodes madagascariensis (Camerano, 1893), Chordodes mobensis Sciacchitano, 1958 and reinterpretation of Chordodes maculatus Sciacchitano, 1958 and Chordodes kakandensis Sciacchitano, 1958

Figure 6. Chordodes maculatus (type series). (A) General view of the midbody cuticle; (B) higher magnification showing simple, thorn, bulging, crowned and circumcluster areoles; (C) areolar cluster with one crowned areole of the female (AMT 32428); (D) longitudinal furrow of the female with crowned areoles with long filaments. Scale bars: 50 µm (A); 20 µm (B, D), 10 µm (C).

Figure 7 in Revision of the genus Chordodes (Gordiida: Nematomorpha) from Africa IV. Ultrastructural redescription of Chordodes congolensis Sciacchitano, 1933, Chordodes ferox Camerano, 1897, Chordodes madagascariensis (Camerano, 1893), Chordodes mobensis Sciacchitano, 1958 and reinterpretation of Chordodes maculatus Sciacchitano, 1958 and Chordodes kakandensis Sciacchitano, 1958

Figure 7. Chordodes kakandensis (holotype). (A) Overview of the midbody cuticle; (B) thorn and tubercle areoles; (C) midline of the body showing crowned areoles with long filaments. Scale bars: 20 µm (A), 10 µm (B), 50 µm (C).

Figure 5 in Revision of the genus Chordodes (Gordiida: Nematomorpha) from Africa IV. Ultrastructural redescription of Chordodes congolensis Sciacchitano, 1933, Chordodes ferox Camerano, 1897, Chordodes madagascariensis (Camerano, 1893), Chordodes mobensis Sciacchitano, 1958 and reinterpretation of Chordodes maculatus Sciacchitano, 1958 and Chordodes kakandensis Sciacchitano, 1958

Figure 5. Holotype of Chordodes mobensis. (A) Cuticle showing simple, bulging and thorn areoles; (B) thorn areole (double arrow) and tubercle areole (white arrow); (C) clusters containing crowned areoles; (D) midline of the body showing crowned areoles with short filaments and thorn areoles. Scale bars: 20 µm (A), 10 µm (B, C).

FIG. 3 in Australian species of Chordodes (Nematomorpha) with a description of two new species, remarks on the genus Chordodes and its life history

FIG. 3. Chordodes queenslandi, female, light microscopy. (A, C) Ventral midline is visible as a dark line. Clusters including areoles of type 5 are distributed along each side of the midline (arrows in A). (B) Cuticle of the lateral side of the body with areoles of types 1, 2 and clusters containing types 4 and 6. (D) Areolar cluster containing types 4 and 5. (E) Areolar types 1–4 and 6 on lateral body.

FIG. 6 in Australian species of Chordodes (Nematomorpha) with a description of two new species, remarks on the genus Chordodes and its life history

FIG. 6. (A–D) Chordodes queenslandi, male, posterior end, SEM. (A) Complete view of the posterior end from the ventral side with bristles close to the cloacal opening (small arrows) and bristleelds anterolateral of the opening (large arrows). (B) Structure of the cuticle in the ventral midline anterior of the cloacal opening. (C) Structure of areoles on the posterior tip of the animal. (D) Structure of cuticle lateral of the cloacal opening with some bristles from the bristleelds (arrows). (E, F) Chordode s queenslandi, specimen from the Zoological Museum Hamburg (V 2231), earlier assigned to C. modiglianii. (E) Areoles type 6 on lateral side of body. (F) Areoles type 5 with long laments in the ventral midline.

FIG. 2 in Australian species of Chordodes (Nematomorpha) with a description of two new species, remarks on the genus Chordodes and its life history

FIG. 2. 'Chordodes ' annulatus (ZMB 4446). Microscopical preparation. (A) Overview of the cuticle. (B) Detail with clustered areoles.

FIG. 8 in Australian species of Chordodes (Nematomorpha) with a description of two new species, remarks on the genus Chordodes and its life history

FIG. 8. Chordodes brevipilus, male posterior end, SEM. (A) Whole posterior end with cloacal opening, lateral bristleŽelds and structurally diVerent regions anterior of the cloacal opening. (B) Ventral midline anterior of the cloacal opening. Note tubercle areoles bordering the midline. (C) BristleŽeld with long bristles (white arrow), spines (black arrow) and minute bristles in between. (D) Spines and cuticular structure in the region posterior of the cloacal opening. (E) Areoles in marginal regions of the bristleŽeld showing minute bristles and tubercle areoles.

FIG. 5 in Australian species of Chordodes (Nematomorpha) with a description of two new species, remarks on the genus Chordodes and its life history

FIG. 5. Chordodes queenslandi, male, light microscopy and SEM. (A) Ventral midline is not as clearly visible as in female. (B, C) Areoles types 2–4 and 6 with light microscopy (B) and SEM (C). (D) Areolar clusters with types 4 and 5. (E) Areoles of type 5. (F, G) Lateral view of type 6 areoles. A central canal is visible (arrows). (H) Areoles type 5.

FIGURE 6 in Novel techniques for biodiversity studies of gordiids and description of a new species of Chordodes (Gordiida, Nematomorpha) from Kenya, Africa

FIGURE 6. Developing and fully formed cysts of Chordodes kenyaensis n. sp. recovered from an experimentally infected Physa gyrina snail. (A) Larva in snail tissue 14 DPE in the process of folding (lateral view). Note the empty pseudointestine (black arrow). Scale bar = 12 µm. (B) Mature cysts (lateral view); note the clear halo surrounding the encysted larvae and single folding pattern. Scale bar = 15 µm.

FIGURE 3 in Novel techniques for biodiversity studies of gordiids and description of a new species of Chordodes (Gordiida, Nematomorpha) from Kenya, Africa

FIGURE 3. Adult and areole pattern of deformed Chordodes kenyaensis n. sp. (A) Adult male removed from a cricket while trying to emerge. Note the larval cuticle (arrow). Scale bar = 1.0 mm. (B) DIC photomicrograph of the mid-body cuticle of a female showing tubercle (2), bulging (4), circumcluster (5) and crowned (6) areoles scattered among simple (1) areoles. Note that simple areoles are smooth but contain short bristles making them appear bumpy. Scale bar = 30 µm. (C) DIC photomicrograph of the same region of the cuticle in a higher focal plane of the mid-body cuticle showing tubercle (2), bulging (4), circumcluster (5) and crowned (6) areoles scattered among simple areoles. Scale bar = 30 µm. (D) DIC photomicrograph of a thorn (3) areole among simple areoles of the mid-body region of an adult deformed female worm. Scale bar = 7 µm.

FIGURE 4 in Novel techniques for biodiversity studies of gordiids and description of a new species of Chordodes (Gordiida, Nematomorpha) from Kenya, Africa

FIGURE 4. Mating, oviposition behavior, egg strings, and eggs of Chordodes kenyaensis n. sp. (A) Male and female in a Gordian knot. Note the lighter color and thicker size of the female worm compared to the male. Scale bar = 2.0 mm. (B) Female in the process of depositing egg strings in a zigzag pattern on a stick. Scale bar = 5 mm. (C) Undeveloped egg. Scale bar = 15 µm.

FIGURE 5 in Novel techniques for biodiversity studies of gordiids and description of a new species of Chordodes (Gordiida, Nematomorpha) from Kenya, Africa

FIGURE 5. Larval characteristics of Chordodes kenyaensis n. sp. (A) DIC photomicrograph of a live larva (lateral view). Note two of the three stylets (black arrow) and v shaped pseudointestine (white arrow). Scale bar = 15 µm. (B) DIC photomicrograph of anterior end of larva (lateral view), note the everted proboscis (arrow) with three stylets. Scale bar = 13 µm. (C) SEM photomicrograph of an entire larva (lateral view). Note the distinct preseptum (Pre) and postseptum (Pos) Scale bar = 8.0 µm. (D) SEM photomicrograph of the posterior end (ventral side). Note the two anterior spines (AS), two posterior spines (PS) and pseudointestine gland opening (PSGO). Scale bar = 3.0 µm. (E) SEM photomicrograph of the anterior end (en face view). Note the orientation of the proboscis (P) with left lateral spines (LLS) to the ventral hooks (VH). Scale bar = 3.0 µm. (F) SEM photomicrograph of the preseptum (ventral view). Note the orientation of the proboscis (P) to the ventral hooks (VH); median hooks (MH). Scale bar = 3.3 µm. (G) Close up SEM photomicrograph of a ventral view of the proboscis (P). Note the arrangement of the five ventral spines (VS). Scale bar = 1.5 µm.

FIGURE 2 in Novel techniques for biodiversity studies of gordiids and description of a new species of Chordodes (Gordiida, Nematomorpha) from Kenya, Africa

FIGURE 2.Allotype of female Chordodes kenyaensis n. sp. (A) SEM photomicrograph showing the tapered anterior end and areole pattern. Note distinct thorn areoles (arrows) among simple areoles. Scale bar = 115 µm. (B) SEM photomicrograph showing higher magnification of the anterior region of the cuticle with simple (1) and tubercle (2) areoles. Note that interareolar furrows are arranged laterally across the cuticle. Scale bar = 10 µm. (C) SEM photomicrograph showing overview of the mid-body cuticle. Note the pattern of crowned areoles. Scale bar = 300 µm. (D) DIC photomicrograph showing two crowned (6) areoles surrounded by circumcluster (5) areoles in the mid-body region. Scale bar = 20 µm. (E) SEM photomicrograph showing simple (1), bulging (4), circumcluster (5) and crown (6) areoles in the mid-body region. Note the long filaments on crown areoles. Scale bar = 20 µm. (F) SEM photomicrograph showing higher magnification of simple (1), tubercle (2), and thorn (3) areoles in the mid-body region. Interareolar furrows are arranged laterally across the cuticle. Note the fine terminal bristles on the simple and tubercle areoles. Scale bar = 10 µm. (G) SEM photomicrograph showing posterior end and cloacal opening (arrow). Note the non-areolated region around the cloacal opening and bristles on the lateral sides of the non-areolated field. Scale bar = 150 µm. (H) SEM photomicrograph showing higher magnification of the posterior region of the cuticle with simple (1), tubercle (2), bulging (4), circumcluster (5) and crowned (6) areoles. Note the variation in simple areoles in this body region compared to the anterior and mid-body region of the cuticle. Scale bar = 40 µm.

FIGURE 1 in Novel techniques for biodiversity studies of gordiids and description of a new species of Chordodes (Gordiida, Nematomorpha) from Kenya, Africa

FIGURE 1. Holotype of male Chordodes kenyaensis n. sp. (A) SEM photomicrograph showing the tapered anterior end and areole pattern. Scale bar = 170 µm. (B) SEM photomicrograph showing higher magnification of the anterior region of the cuticle with simple (1), tubercle (2), and two types of thorn (3 and 3*) areoles. Note that interareolar furrows are arranged laterally across the cuticle. Scale bar = 15 µm. (C) SEM photomicrograph showing an overview of the mid-body cuticle. Note the pattern of crowned areoles. Scale bar = 190 µm. (D) SEM photomicrograph showing a higher magnification of the midbody cuticle containing simple (1), tubercle (2), thorn (3), bulging (4), circumcluster (5) and crowned (6) areoles. Interareolar furrows are arranged laterally across the cuticle. Note the long filaments on crowned areoles and short bristles on top of simple smooth areoles which are separated between neighbors by interareolar furrows. Scale bar = 25 µm. (E) SEM photomicrograph showing the posterior end and cloacal opening (arrow). Note the indistinct lobes (thick arrows). Scale bar = 115 µm. (F) SEM photomicrograph showing a higher magnification of the posterior region of the cloacal opening surrounded by circumcloacal spines. Note the non-areolated region and bristles on the lateral regions of the non-areolated field. Scale bar = 40 µm.

FIGURE 7 in Novel techniques for biodiversity studies of gordiids and description of a new species of Chordodes (Gordiida, Nematomorpha) from Kenya, Africa

FIGURE 7. Molecular phylogenetic analysis by maximum likelihood method based on partial cox1 gene from Chordodes kenyaensis n. sp. and 16 other nucleotide sequences from Genbank. Paragordius and Gordius were used as outgroups. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. Only bootstrap values with a value of 85% or higher are shown. Note that Chordodes kenyaensis n. sp. is placed within other species of Chordodes.