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Figure 3 from: Nielsen SV, Conradie W, Ceríaco LMP, Bauer AM, Heinicke MP, Stanley EL, Blackburn DC (2020) A new species of Rain Frog (Brevicipitidae, Breviceps) endemic to Angola. ZooKeys 979: 133-160. https://doi.org/10.3897/zookeys.979.56863
Figure 3 Spectrograms and oscillograms showing a series of notes of the putatively novel Angolan Breviceps taxon compared to three closely related congeners.
Figure 1 from: Nielsen SV, Conradie W, Ceríaco LMP, Bauer AM, Heinicke MP, Stanley EL, Blackburn DC (2020) A new species of Rain Frog (Brevicipitidae, Breviceps) endemic to Angola. ZooKeys 979: 133-160. https://doi.org/10.3897/zookeys.979.56863
Figure 1 Geographic distribution and phylogenetic relationships of Breviceps spp. included in this study. A Map of Angola and surrounding countries with all known Breviceps spp. sampling localities indicated on legend. The proposed distributions of B. adspersus and B. poweri (blue and red polygons, respectively) are from IUCN (2013a, b), but should be considered tentative and worthy of reevaluation in light of recent studies. Furthermore, B. mossambicus is not mapped as no samples of certain identification occur west of Malawi (see Nielsen et al. 2018). B Multi-locus phylogeny of Breviceps, with select clades collapsed that are not relevant directly to the B. mossambicus group. The backbone is from the likelihood analysis, although Bayesian analyses produced a nearly identical topology (with any topological differences subtended by poor support). A black dot at each node indicates high support (e.g., Bayesian posterior probability > 0.95, Maximum Likelihood bootstrap > 90), while values below that cutoff are indicated for deep nodes only. Tapered bars to the right of voucher IDs indicate from which Angolan locality they were collected. C Median-joining networks for the two nuclear loci indicating a lack of shared haplotypes between candidate and recognized species. Hash marks indicate unique sequence differences between lineages, and black circles are hypothetical intermediate haplotypes.
Figure 5 from: Nielsen SV, Conradie W, Ceríaco LMP, Bauer AM, Heinicke MP, Stanley EL, Blackburn DC (2020) A new species of Rain Frog (Brevicipitidae, Breviceps) endemic to Angola. ZooKeys 979: 133-160. https://doi.org/10.3897/zookeys.979.56863
Figure 5 Variation in color and pattern within living paratypes of B. ombelanonga sp. nov.: A, B sub-adult (of unknown sex) from Embala Seque (14 km N of Cassumbi village), Bié Province (MHNCUP_ANF 0320) C juvenile male, Cuito River source lake, Moxico Province (PEM A12537) D adult female, Cuando River source, Moxico Province (PEM A12770) E adult male, Quembo River source lake, Moxico Province (PEM A12787) F adult male, Cuanavale River source lake, Moxico Province (PEM A12800). Photographs by LMPC (A, B) and WC (C–F).
Supplementary material 1 from: Nielsen SV, Conradie W, Ceríaco LMP, Bauer AM, Heinicke MP, Stanley EL, Blackburn DC (2020) A new species of Rain Frog (Brevicipitidae, Breviceps) endemic to Angola. ZooKeys 979: 133-160. https://doi.org/10.3897/zookeys.979.56863
Table S1. Morphological data used to perform PCAs
Figure 2 from: Nielsen SV, Conradie W, Ceríaco LMP, Bauer AM, Heinicke MP, Stanley EL, Blackburn DC (2020) A new species of Rain Frog (Brevicipitidae, Breviceps) endemic to Angola. ZooKeys 979: 133-160. https://doi.org/10.3897/zookeys.979.56863
Figure 2 PCA plots of ten size-corrected morphological characters from specimens of B. adspersus (n = 24), B. mossambicus (n = 9), B. poweri (n = 8), and the putative new Angolan species (n = 6) (Suppl. material 1: Table S1), illustrating the PC1 and PC2 (A) and PC2 and PC3 (B) axes of variation, which combined represent ~ 70% of the total variation (Table 4).
Figure 7 from: Nielsen SV, Conradie W, Ceríaco LMP, Bauer AM, Heinicke MP, Stanley EL, Blackburn DC (2020) A new species of Rain Frog (Brevicipitidae, Breviceps) endemic to Angola. ZooKeys 979: 133-160. https://doi.org/10.3897/zookeys.979.56863
Figure 7 Photos of typical habitat of B. ombelanonga sp. nov.: A a view of the Kwanza River and bordering savannah, near the type locality, in Kissama National Park, Luanda Province B savannah near Embala Seque (14 km N of Cassumbi village), Bié Province C Cuanavale River source lake and associated miombo savannah woodland. Photographs by LMPC (A, B) and WC (C).
Figure 4 from: Nielsen SV, Conradie W, Ceríaco LMP, Bauer AM, Heinicke MP, Stanley EL, Blackburn DC (2020) A new species of Rain Frog (Brevicipitidae, Breviceps) endemic to Angola. ZooKeys 979: 133-160. https://doi.org/10.3897/zookeys.979.56863
Figure 4 Breviceps ombelanonga sp. nov. holotype male (UF Herp 187172): A in life photo B dorsal and ventral aspects C additional views of the holotype, including the left pes, frontal, right lateral, and left manus and mental. Scale bar: 10 mm. Photographs by J. Cavagnaro (A) and SVN (B, C).
Figures 32- 33 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figures 32- 33 Principal component analysis (PCA) and Redundancy analysis (RDA) with species or molecular lineage constraint applied to the original shell matrix (32) and shape-related Z-matrix (33).
Figures 23-31 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figures 23-31 Shell variability in Monacha pantanellii from Via Salaria, Poggio San Lorenzo [Lor] (FGC 41551) (23–25), Valle dell'Aniene, Roccagiovine [Ani] (FGC 42974) (26–29) and Carsoli [Car] (FGC 41651(30, 31).
Figure 34 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figure 34 Box plots for shell characters of the nine Monacha species or molecular lineages investigated. The lower and upper limits of the rectangular boxes indicate the 25th to 75th percentile range, and the horizontal line within the boxes is the median (50th percentile).
Figure 4 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figure 4 Bayesian 50% majority-rule consensus tree of the concatenated data set of COI and 16S rDNA haplotypes, and ITS2 and H3 common sequences (see Table 4). Sequences of M. pantanellii were compared with appropriate sequences of M. cantiana s. l. and M. parumcincta obtained from GenBank (Tables 2, 4). Posterior probabilities (left) and bootstrap support above 50% from ML analysis (right) are indicated next to the branches. Bootstrap analysis was run with 1000 replicates (Felsenstein 1985). The tree was rooted with M. cartusiana concatenated sequences obtained from GenBank (Table 2).
Figures 15-22 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figures 15-22 Shell variability in Monacha pantanellii from Lago del Turano [Tur2] (FGC 41654) (15–17), Via Salaria, Ornaro Alto [Alt] (FGC 41553) (18–20) and Vallonina [Val] (FGC 25345) (21, 22).
Figures 5-14 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figures 5-14 Shell variability in Monacha pantanellii from Monte Fionchi, summit [Fio1] (FGC 8140) (5, 6), Monte Fionchi, Torrecola [Fio2] (FGC 38944) (7), road to Montenero Sabino [Sab] (FGC 41552) (8–10) and Turania [Tur1] (FGC 42971) (11–14).
Figure 3 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figure 3 Maximum Likelihood (ML) tree of concatenated ITS2 and H3 common sequences of Monacha pantanellii (see Table 4). New ITS2 and H3 sequences of M. pantanellii (Table 1) were compared with ITS2 and H3 sequences of M. cantiana s. l. and M. parumcincta obtained from GenBank (Tables 2, 4). Numbers next to branches indicate bootstrap support above 50% calculated on 1000 replicates (Felsenstein 1985). The tree was rooted with M. cartusiana concatenated sequences obtained from GenBank (Table 2).
Figure 66 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figure 66 Box plots for genital characters of the ten Monacha species or molecular lineages investigated. The lower and upper limits of the rectangular boxes indicate the 25th to 75th percentile range, and the horizontal line within the boxes is the median (50th percentile).
Figures 67- 68 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figures 67- 68 Principal component analysis (PCA) and Redundancy analysis (RDA) with population constraint applied to the original genital matrix (67) and shape-related Z-matrix (68) of specimens of Monacha pantanellii.
Figures 60-63 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figures 60-63 Internal structure of distal genitalia of Monacha pantanellii from Valle dell'Aniene, Roccagiovine [Ani] (FGC 42974) (60), Via Salaria, Ornaro Alto [Alt] (FGC 41553) (61), Turania [Tur1] (FGC 42971) (62) and road to Montenero Sabino [Sab] (FGC 41552) (63).
Figures 49-52 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figures 49-52 Genitalia (proximal parts excluded) (49), internal structure of distal genitalia (50), transverse sections of medial epiphallus (51) and apical penial papilla (52) of Monacha pantanellii from Valle dell'Aniene, Roccagiovine [Ani] (FGC 42974).
Figures 45-48 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figures 45-48 Genitalia (proximal parts excluded) (45), internal structure of distal genitalia (46), transverse sections of medial epiphallus (47) and apical penial papilla (48) of Monacha pantanellii from Vallonina [Val] (FGC 25345).
Figures 35- 36 from: Pieńkowska JR, Manganelli G, Giusti F, Barbato D, Kosicka E, Hallgass A, Lesicki A (2020) Redescription of Monacha pantanellii (De Stefani, 1879), a species endemic to the central Apennines, Italy (Gastropoda, Eupulmonata, Hygromiidae) by an integrative molecular and morphological approach. ZooKeys 988: 17-61. https://doi.org/10.3897/zookeys.988.56397
Figures 35- 36 Principal component analysis (PCA) and Redundancy analysis (RDA) with population constraint applied to the original shell matrix (35) and shape-related Z-matrix (36) of specimens of Monacha pantanellii.
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