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FIGURE 5 in A new frog species from rapidly dwindling cloud forest streams of Sri Lanka-Lankanectes pera (Anura, Nyctibatrachidae)

FIGURE 5. Cleared and stained adult specimen of Lankanectes corrugatus (DZ1397, SVL 45.02 mm). (A) Cranium, dorsal view. (B) Vertebral column, with the pelvic girdle attached to the sacral diapophysis via the cartilaginous epiphysis of ilia, dorsal view. (C) Close-up of vertebrae (D) Hyobranchial skeleton, ventral view. (E) Pectoral girdle and forelimbs, ventral view. Abbreviations: CR, coracoid; CT, cleithrum; CV, clavicle; EX, exoccipital; FP, frontoparietal; HP, hyoid plate; HY, hyale; IL, ilium; IS, ischium; MX, maxillae; OS, omosternum; PM, premaxilla; PP, posteromedial process; PR, prezygapophysis; PZ, postzygapophysis; PT, pterygoid; SC, suprascapula; SQ, squamosal; SL, scapula; ST, sternum; PV, vomer.

FIGURE 2 in A new frog species from rapidly dwindling cloud forest streams of Sri Lanka-Lankanectes pera (Anura, Nyctibatrachidae)

FIGURE 2. Lankanectes pera sp. nov. in life, SVL 22.40 mm: (A) Dorsolateral view; (B) Dorsal view; (C) Habitat—clear water stream habitat under montane forest canopy cover.

FIGURE 4 in A new frog species from rapidly dwindling cloud forest streams of Sri Lanka-Lankanectes pera (Anura, Nyctibatrachidae)

FIGURE 4. Lankanectes pera sp. nov., SVL 68.69 mm, DZ1307 (A–D), in preservation. (A) ventral view; (B) right ventral foot; (C) ventral view of the head; (D) lateral view. Lankanectes corrugatus, SVL 55.28 mm, DZ1396 (E–H), in preservation. (A) ventral view; (B) right ventral foot; (C) ventral view of the head; (D) lateral view.

FIGURE 1 in A new frog species from rapidly dwindling cloud forest streams of Sri Lanka-Lankanectes pera (Anura, Nyctibatrachidae)

FIGURE 1. (A) 16S rRNA Maximum-likelihood phylogram for 41 taxa with node support (posterior probabilities above nodes, with values>95 indicated by a *; MP bootstrap values below nodes, with values>50) showing two clearly-defined clades including L. corrugatus and L. pera (B) The uncorrected pairwise genetic distances range for 16S rRNA 3.5–3.7%; Haplotype network analysis using the full dataset with 485 bp, shows all haplotypes shared across L. corrugatus; L. pera sp. nov. is different by more than 16 mutational steps from the L. corrugatus populations. (C) The predicted distributions for L. corrugatus is 360 km2 and L. pera sp. nov. is 14120 km2 on the northern hills of Sri Lanka. (D) Component loadings of the first two principal component axes (PC1 vs PC2) shows clear separation of the males, but with slight overlap of females; L. pera sp. nov. indicated in red, L. corrugatus in blue (filled circles denote females and open circles denote males).

FIGURE 3 in A new frog species from rapidly dwindling cloud forest streams of Sri Lanka-Lankanectes pera (Anura, Nyctibatrachidae)

FIGURE 3. Illustrations of (A) Lankanectes corrugatus, ZMB4897, mature male, SVL 43.70 mm and (B) L. pera sp. nov., DZ1858, mature male, SVL 66.0 mm, anal region (top); manus (middle) and maxilla (lower).

Figure 4 in A new ancient lineage of frog (Anura: Nyctibatrachidae: Astrobatrachinae subfam. nov.) endemic to the Western Ghats of Peninsular India

Figure 4 Paratype of Astrobatrachus kurichiyana- ZSI/WRC/A/2133 (female): Ventral head (A), dorsal head (B), ventral (C), ventral fingers (D), lateral head (E), ventral toes (F). Drawing Credit: Achyuthan N. Srikanthan Full-size DOI: 10.7717/peerj.6457/fig-4

Figure 3 in A new ancient lineage of frog (Anura: Nyctibatrachidae: Astrobatrachinae subfam. nov.) endemic to the Western Ghats of Peninsular India

Figure 3 Live images of Astrobatrachus kurichiyana. Profile (A), close-up of head (B), ventral (C), dorsal (D), side-profile (E). Photo Credit: S.P. Vijayakumar (A and B; reference collection CESF 1567), K.P. Dinesh (C, D and E; ZSI/WRC/A/2131) Full-size DOI: 10.7717/peerj.6457/fig-3

Figure 1 in A new ancient lineage of frog (Anura: Nyctibatrachidae: Astrobatrachinae subfam. nov.) endemic to the Western Ghats of Peninsular India

Figure 1 Geographical range (A) of the three genera, Nyctibatrachus (Nyctibatrachinae), Lankanectes (Lankanectinae) and the new genus Astrobatrachus (Astrobatrachinae subfam. nov.). Inset maps show the type locality (B) and the narrow range (C) of Astrobatrachus kurichiyana gen et. sp. nov. Photo Credit: S. P. Vijayakumar. Full-size DOI: 10.7717/peerj.6457/fig-1

Figure 6 in A new ancient lineage of frog (Anura: Nyctibatrachidae: Astrobatrachinae subfam. nov.) endemic to the Western Ghats of Peninsular India

Figure 6 Comparisons of male paratype (ZSI/WRC/2132) of Astrobatrachus kurichiyana to representatives of other WG endemics. Exemplars from Nyctibatrachidae, Micrixalidae, and Ranixalidae in different views ((A–F) entire skeleton in dorsal view; (G–L) dorsal view of skull; (M–R) ventral view of skull; (S–X) ventral view of pectoral girdle). Abbreviations of anatomical terms are as follows: av, anterior vomer; clav, clavicle; np, neopalatine; omo, omosternum; pv, posterior vomer. Full-size DOI: 10.7717/peerj.6457/fig-6

FIGURE 11 in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 11. Female and Male SVL (Mean±Sd, in mm) and clutch size (Mean±Sd) in three allopatric (N. humayuni, N. petraeus and N. jog) and three sympatric (N. jog, N. kempholeyensis and Nyctibatrachus kumbara sp. nov.) species of Nyctibatrachus.

FIGURE 8 in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 8. Nyctibatrachus kumbara sp. nov. in axillary amplexus. Adult male (Holotype ZSI/WGFRS/V/A/860) on top of adult female in swampy evergreen forest of Kathalekan, Uttara Kannada district, Karnataka, India. Female not collected.

FIGURE 10 in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 10. Co-occurring congeneric (COCG) species of Nyctibatrachus in Kathalekan. a. Nyctibatrachus kempholeyensis, b. N. jog and c. Nyctibatrachus kumbara sp. nov.

FIGURE 7. Maximum likelihood tree for all 28 in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 7. Maximum likelihood tree for all 28 nominal species of Nyctibatrachus and an outgroup (Indirana sp.) based on mitochondrial 16S rRNA. Number at the branches indicate bootstrap values. Bootstrap values less than 50 are indicated with an asterisk. Area marked with grey belong to N. sanctipalustris clade.

FIGURE 6. a in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 6. a. Male approaching the eggs; b. Male standing upright locating the egg clutch; c. Male covering the egg clutch with sand gravels and mud from the stream bed; d. Male leaving the place after covering the egg mass with sand and mud.

FIGURE 5. a in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 5. a. Male and female in courtship (male and female are marked with symbols); b. Axillary amplexus in the pair; c. Amplected pair standing upright on their hindlimb before oviposition; d. Female doing a handstand at the time of releasing eggs.

FIGURE 4b. Type II in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 4b. Type II advertisement call spectrogram of Nyctibatrachus kempholeyensis. a. Amplitude and b. Spectrogram.

FIGURE 1 in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 1. Study localities (blue circles) of Nyctibatrachus kumbara sp. nov. in the Western Ghats of India.

FIGURE 4a. Type I in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 4a. Type I advertisement call spectrogram of Nyctibatrachus kempholeyensis. a. Amplitude and b. Spectrogram.

FIGURE 9 in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 9. Holotype (ZSI/WGFRS/V/A/860) of Nyctibatrachus kumbara sp. nov. A—Dorsal view; B—Ventral View; C—Lateral profile of head; D—Ventral view of Forelimb; E—Ventral view of Hindlimb; F—Dorsal view of third finger; G—Dorsal view of fourth toe and H—Schematic view of webbing in hindlimb.

FIGURE 12 in Mud-packing frog: A novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae)

FIGURE 12. Non-metric multidimensional scale (NMDS) analysis of allopatric and sympatric species of Nyctibatrachus based on SVL (both male and female), maximum number of eggs in a clutch, type of amplexus, parental care, mud covering behavior, oviposition site and position of male and female at oviposition. Allopatric species namely N. humayuni, N. petraeus and N. jog are clustered in one group, while N. kempholeyensis and Nyctibatrachus kumbara sp. nov. forms two distinct clusters.