Journal of Natural History Vol. 45, Nos. 9–10, March 2011, 521–552 Morphometric and molecular variation in mountain catfishes (Amphiliidae: Amphilius) in Guinea, West Africa Ray C. Schmidta,b∗ and Frank Pezolda a Department of Life Sciences, Texas A&M University Corpus Christi, Corpus Christi, TX, USA; Present affiliation: Department of Ecology and Evolutionary Biology, Tulane University, New Orleans, LA, USA b (Received 31 August 2009; final version received 19 October 2010; printed 28 January 2011) Species of the genus Amphilius are small benthic catfishes that occur throughout tropical Africa and are abundant in the mountain streams of the Fouta Djalon region in Guinea, West Africa. During a 2003 expedition to the area, numerous amphiliid catfish were collected and initial observations revealed a large amount of morphological variation across the sampled basins. These specimens were subjected to a geometric morphometric analysis and standard morphometric analysis. Cytochrome b gene sequences were also obtained and compared among populations. These analyses resulted in the discovery of multiple cryptic taxa within two recognized species, Amphilius platychir and Amphilius rheophilus. The discovery of these taxa is compared with other recent discoveries from the area. Keywords: Amphilius platychir; Amphilius rheophilus; geometric morphometrics; cytochrome b; Fouta Djalon Introduction There are currently 28 members of the catfish genus Amphilius that inhabit fast moving streams throughout tropical Africa. One such area is the Fouta Djalon region in Guinea, West Africa. These highlands rise over 1000 m above sea level and serve as a headwater source to several major rivers of West Africa – the Senegal, the Gambie, the Corubal and the Tinkisso, a tributary of the upper Niger. The highlands separate the grasslands and brush country of the Sahelian interior from the tropical rainforests along the coast. The Fouta Djalon region is also home to a large number of endemic species (Hugueny and Lévêque 1994). Amphilius species are well-adapted to life within the high-gradient Fouta Djalon streams. Their bodies are flattened dorsoventrally and have large pectoral fins with stiffened first rays that allow them to cling to the benthic substrate in fast-flowing rivers. The Fouta Djalon region, which lies within the Upper Guinean ichthyofaunal province (Roberts 1975), remains one of the least explored areas in West Africa (Teugels et al. 1987). The first substantial survey of the area’s ichthyofauna was reported by Daget (1962) who listed 97 species from the Fouta Djalon, 15 of which were previously undescribed species. Subsequent surveys of the area have produced numerous new species, most of which are endemic to the region (Teugels et al. 1987; Howes and Teugels 1989; Ramond 1994; Vreven and Teugels 2005). *Corresponding author. Email: rschmidt@tulane.edu ISSN 0022-2933 print/ISSN 1464-5262 online © 2011 Taylor & Francis DOI: 10.1080/00222933.2010.534560 http://www.informaworld.com 522 R.C. Schmidt and F. Pezold Amphilius platychir Günther 1864, was first described from Sierra Leone. This type locality was questioned by Boulenger (1898) as he placed A. platychir in East Africa. Although Günther (1902) and Poche (1902) argued the veracity of the type locality, Boulenger (1911) continued to regard the locality as suspect. Boulenger’s assertion that A. platychir was an East African species may have encouraged subsequent descriptions of additional Amphilius species and subspecies from West Africa (Pellegrin 1913, 1935; Daget 1963). In a revision of the eastern and southern African species of the genus, Skelton (1984) corrected Boulenger (1898), recognizing Sierra Leone as the type locality of A. platychir, and placed a number of species as junior synonyms of A. platychir. Currently, A. platychir, Amphilius rheophilus Daget 1959, Amphilius atesuensis Boulenger 1904 and Amphilius kakrimensis Teugels et al. 1987 are the only recognized species of Amphilius recorded within Guinea, West Africa (Paugy et al. 2003) (Figure 1). Body proportions and features of the head are useful in identifying Amphilius species from the Fouta Djalon region. Amphilius atesuensis is easily distinguished from other amphiliids in having its dorsal fin terminus placed above or posterior to its pelvic fin insertion (Paugy et al. 2003). Amphilius rheophilus, described from the Gambie River in Senegal, is generally identified as having a caudal peduncle length-to-height ratio greater than 2.0, shorter maxillary barbels (65% of head length), and long snout (greater than 50% of head length) (Paugy et al. 2003). Amphilius platychir is identified by long maxillary barbels (over 75% of head length), a caudal peduncle length-to-height ratio of 1.5–2.0, and a shorter snout (less than 50% of head length) (Paugy et al. 2003). Amphilius kakrimensis, described from the Kakrima River, is said to have a deeper body and caudal peduncle and eyes that are set further apart than other amphiliids (Teugels et al. 1987). Colouration is also useful in distinguishing species as A. atesuensis and A. rheophilus display dorsal saddles, and A. platychir and A. kakrimensis are characterized by light horizontal bands above and below the lateral line (Paugy et al. 2003). Although fin ray number is often helpful in identifying fish species, counts for these taxa are generally similar. During several expeditions to the Fouta Djalon and surrounding localities in 2003, amphiliid catfish were collected from major basins throughout the region. Initial investigation of the specimens suggested that variation in body shape morphology occurred across the various basins. This study quantifies and analyses morphological variation among putative Amphilius species and examines its concordance with genetic diversity. Materials and methods Specimens for morphological analysis were collected at 47 sites throughout Guinea, West Africa from January through to May 2003 (Figure 2). The collections were focused on the Fouta Djalon region, upper Niger River basin, Zone Forestière, and coastal rivers. Specimens collected with seines and dip-nets were fixed with 10% formalin and preserved in 50% isopropyl alcohol. These 47 collection sites lie within 11 basins: Senegal, Gambie, Rio Corubal, Tinguilinta, Fatala, Konkouré, Kolenté, Little Scarcies, Loffa, St Paul and Niger. Type specimens of A. rheophilus, A. platychir, Amphilius grammatophorus, A. grammatophorus inequalis, A. kakrimensis, and additional non-type materials from Liberia and Sierra Leone were also examined. All specimens examined are listed with institutional abbreviations following Leviton Journal of Natural History 523 Figure 1. The four Amphilius species currently recognized from Guinea. (A) Amphilius platychir from the Little Scarcies basin; (B) A. atesuensis from the St Paul River basin; (C) A. rheophilus from the Rio Corubal basin; and (D) paratype of A. kakrimensis (MNHN 1986-600). Scale bar is 1 cm. et al. (1985) except for field numbers ABS and FP03 which will be catalogued before the complete systematic revision (Appendix 1). Amphilius atesuensis specimens were not included in the morphological analysis because they are morphologically distinct from other West African amphiliid species in the position of dorsal fin (Paugy et al. 2003). To quantify morphological variation, both geometric morphometric features and standard linear measurements were analysed. 524 R.C. Schmidt and F. Pezold Figure 2. Locations where amphiliid specimens (circle) and tissue samples (star) were collected during the 2003 expedition. Geometric morphometrics Geometric morphometric analysis (GM) is increasingly being used to assess intraspecific variation (Barluenga et al. 2006; Hankison et al. 2006; Reis et al. 2006). GM has also been successful in evaluating the variation that occurs among and between cryptic taxa (Valentin et al. 2002; Guill et al. 2003; Reis et al. 2006). GM techniques have often been shown to be more informative in analysing and interpreting body form variations than standard techniques (e.g. Parsons et al. 2003). Lateral, dorsal, ventral images were acquired for each specimen with a Sony DSC-R1 camera mounted on a copy stand. Deformed specimens were not included in the GM. To aid in the location of landmarks when digitizing, insect mounting pins were placed at landmark locations that were difficult to view after digitization. The landmarks used in the GM are comparable to homologous structures (Humphries 2002) (Figure 3). Landmarks were digitized using TPSDIG ver. 1.40 software (Rohlf 2004). Nonshape variation as the result of size, orientation and location was removed using the Generalized Procrustes Superimposition (Rohlf and Slice 1990) using COORDGEN6 [by H.D. Sheets, available as part of the INTEGRATED MORPHOMETRICS PACKAGE (IMP) at: http://www2.canisius.edu/∼sheets/ morphsoft.html]. PCAGEN6 (IMP) was used to calculate partial warp scores and Journal of Natural History 525 Figure 3. Generalized Amphilius with location of landmarks used in geometric analysis for (A) lateral, (B) dorsal and (C) ventral analysis. these scores were used to perform a principal components analysis. Implementing a principal components analysis provides an analysis without any a priori assumptions of groups, which allows for the unbiased discovery of such groups. Significant shape differences between and within species were assessed with a bootstrapped Goodall’s F-test (Goodall 1991) in TWOGROUP6A (IMP). Body shape change between species and populations inhabiting different basins was visualized using thin-spline deformation grids using TWOGROUP6A (IMP). Standard morphometrics Analyses of standard morphometric features were performed because some structures do not lend themselves well to GM (Bookstein 1991). For example, GM requires that landmarks be fixed points. As barbels are free to rotate around their insertion, it would be exceedingly difficult to include barbel length in the geometric analysis. These analyses also allowed us to quantify differences among populations for specific morphological features proposed as diagnostic for Amphilius species. Linear measurements were taken from the left side of specimens to the nearest 0.1 mm with digital callipers following the methods of Skelton (1981). Head length (HL), predorsal length (PrDL), adipose fin length (AL), caudal peduncle length (CL) and caudal peduncle height (CH) were expressed as percentages of standard length (SL). Maxillary barbel length (BL), interorbital distance (IO) and snout length (SnL) were noted as a percentage of head length (HL). Caudal peduncle length and caudal peduncle height were also expressed as the ratio CL/CH. All ratios were log-transformed before univariate analyses were executed in SYSTAT 12.0 (SYSTAT Software Inc., Chicago, IL, USA; 2007). A principal component analysis on the linear measurements was also performed and plotted by species and basins using SYSTAT 12.0. Gill raker counts were taken from the first gill arch on the left side. 526 R.C. Schmidt and F. Pezold Cytochrome b analysis Molecular analyses are also very useful in discovering cryptic taxa (Murphy and Collier 1996; Near et al. 2000; Egge and Simmons 2006). One of the most widely used molecular markers for fish is the mitochondrial cytochrome b gene. To date, no molecular studies have been performed on amphiliid catfish of the Fouta Djalon region. Tissue samples of 52 Amphilius specimens were collected from 10 sites within the study area (Figure 1) and immediately preserved in 95% ethanol. The samples were stored at – 80◦ C on return to the laboratory. Voucher specimens were retained for each tissue sample, fixed with 10% formalin and preserved in 50% isopropyl alcohol (Table 1). Table 1. Locality, GenBank accession numbers, museum catalogue numbers and tissue numbers for voucher specimens used in molecular analysis. All specimens collected in Guinea, West Africa. Species Locality GenBank No. Catalogue No. Amphilius platychir Fatala River basin, Boffa, Koumbouya River at Bakoro Bridge Fatala River basin, Boffa, Koumbouya River at Bakoro Bridge Fatala River basin, Boffa, Koumbouya River at Bakoro Bridge Fatala River basin, Boffa, Koumbouya River at Bakoro Bridge Rio Corubal basin, Gaoual, Senta River at Senta Bridge Rio Corubal basin, Gaoual, Senta River at Senta Bridge Rio Corubal basin, Gaoual, Senta River at Senta Bridge Rio Corubal basin, Gaoual, Senta River at Senta Bridge Rio Corubal basin, Gaoual, Senta River at Senta Bridge GQ379946 AMNH 250572 RS1 GQ379975 AMNH 250573 RS2 GQ379976 AMNH 250574 RS3 GQ379977 AMNH 250575 RS4 GQ379978 AMNH 250576 RS5 GQ379979 AMNH 250577 RS6 GQ379980 AMNH 250578 RS7 GQ379981 AMNH 250579 RS8 GQ379947 AMNH 250580 RS9 Amphilius platychir Amphilius platychir Amphilius platychir Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Tissue No. (Continued) Journal of Natural History 527 Table 1. (Continued). Species Locality GenBank No. Catalogue No. Amphilius platychir Senegal River basin, Dalaba, Téné River at Thianguelnoussi Senegal River basin, Dalaba, Téné River at Thianguelnoussi Senegal River basin, Dalaba, Téné River at Thianguelnoussi Senegal River basin, Dalaba, Téné River at Thianguelnoussi Senegal River basin, Dalaba, Téné River at Thianguelnoussi Rio Corubal basin, Gaoual, Finton River at Bridge at Dandoun (Kembra) Rio Corubal basin, Gaoual, Finton River at Bridge at Dandoun (Kembra) Rio Corubal basin, Gaoual, Finton River at Bridge at Dandoun (Kembra) Rio Corubal basin, Gaoual, Finton River at Bridge at Dandoun (Kembra) Gambie River basin, Tougue, Dimma River at Diogoma Gambie River basin, Tougue, Dimma River at Diogoma Gambie River basin, Tougue, Dimma River at Diogoma Gambie River basin, Tougue, Dimma River at Diogoma GQ379982 AMNH 248683 3415T GQ379983 AMNH 248683 3413T GQ379948 AMNH 250581 11503O GQ379944 AMNH 248695 3102T GQ379945 AMNH 248695 3414T GQ379949 AMNH 248708 1465T GQ379950 AMNH 248708 1466T GQ379984 AMNH 250583 RS10 GQ379951 AMNH 250584 RS11 GQ379985 AMNH 250585 11203D GQ379952 AMNH 250587 3114T GQ379986 AMNH 250588 3115T GQ379953 AMNH 250589 11203F Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) GQ379954 AMNH 250590 11303C Amphilius platychir Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Tissue No. (Continued) 528 R.C. Schmidt and F. Pezold Table 1. (Continued). Species Locality GenBank No. Catalogue No. Amphilius platychir Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Fétoré River at Wassa (Tinbitounni) Konkouré River basin, Pita, Saalawol River at Lannou Senegal River basin, Mamou, Poukouwol River at Timbo-Niagara Senegal River basin, Mamou, Poukouwol River at Timbo-Niagara GQ379955 AMNH 250591 11303D GQ379942 AMNH 250592 11303E GQ379943 AMNH 250593 11303F GQ379956 AMNH 250594 11303G GQ379957 AMNH 250595 11303H GQ379958 AMNH 250596 11303I GQ379959 AMNH 250596 11303J GQ379960 AMNH 250600 11303N GQ379987 AMNH 250703 11303O GQ379961 AMNH 250704 11303P GQ379962 AMNH 250705 11303Q GQ379963 AMNH 250706 11303R GQ379966 AMNH 250707 11303AY GQ379967 AMNH 250708 11603P GQ379968 AMNH 250709 11603Q Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius rheophilus Amphilius rheophilus Tissue No. (Continued) Journal of Natural History 529 Table 1. (Continued). Species Locality GenBank No. Catalogue No. Amphilius rheophilus Senegal River basin, Mamou, Bafing River at Basambaya Senegal River basin, Mamou, Bafing River at Basambaya Senegal River basin, Mamou, Bafing River at Basambaya Senegal River basin, Mamou, Bafing River at Basambaya Senegal River basin, Mamou, Bafing River at Basambaya Senegal River basin, Mamou, Bafing River at Basambaya Konkouré River basin, Pita, Saalawol River at Lannou Konkouré River basin, Pita, Saalawol River at Lannou Konkouré River basin, Pita, Saalawol River at Lannou Konkouré River basin, Pita, Saalawol River at Lannou Konkouré River basin, Pita, Saalawol River at Lannou Konkouré River basin, Pita, Saalawol River at Lannou Konkouré River basin, Pita, Saalawol River at Lannou St. Paul River basin, Nzérékoré, Loh River at Gnampara Kolenté River basin, Kindia, Sole River at Galeya GQ379988 AMNH 250710 3497T GQ379989 AMNH 250711 3498T GQ379990 AMNH 250712 3495T GQ379969 AMNH 250713 3500T GQ379970 AMNH 250715 11803F GQ379991 AMNH 250716 11803L GQ379971 AMNH 250717 11303AZ GQ379972 AMNH 250718 11303BA GQ379973 AMNH 250720 11303BC GQ379974 AMNH 250721 11303BD GQ379964 AMNH 250722 11303BE GQ379965 AMNH 250723 11303BF GQ379992 AMNH 250724 11303BG GQ379993 AMNH 248702 T5658 GQ379941 AMNH 250725 3459T Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius rheophilus Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius platychir Amphilius rheophilus Amphilius atesuensis Malapterurus electricus Tissue No. 530 R.C. Schmidt and F. Pezold Total genomic DNA from each specimen was extracted with the DNeasy Tissue Kit (Qiagen, Valencia, CA, USA) following the manufacturer’s protocol. Partial cytochrome b sequences were amplified from the genomic DNA using the polymerase chain reaction (PCR). The primers used were GLUDGL (5′ -TGA CCT GAA RAA CCA YCG TTG-3′ ) and CB3H (5′ -GGC AAA TAG G AA RTA TCA TTC-3′ ) (Palumbi 1996). The 25-µl PCR contained approximately 100 ng template DNA, 1 µl of each primer (10 mM), 5 µl of 5× reaction buffer, 1 µl MgCl2 (25 mM), 0.5 µl dNTP (10 mM), 1.5 units of GoTaq (Promega Inc., Madison, WI, USA), and 15.7 µl sterile water. The reactions were cycled at the following temperatures: an initial 94◦ C denaturation for 5 min, then 40 cycles of 94◦ C denaturation for 1 min, 48◦ C annealing for 1 min, 72◦ C extension for 1 min, and a final 72◦ C extension for 7 min. The PCR products were purified with QiaQuick PCR purification kit (Qiagen) and sequenced using a GenomeLab DTCS Quick Start (Beckman Coulter, Brea, CA, USA) kit with amplification primers (GLUDGL and CB3H) on a CEQ 8000 sequencer (Beckman Coulter) Multiple sequence alignment was performed with CLUSTAL W ver. 1.4 (Thompson et al. 1994); sequences were edited in BIOEDIT ver. 7.0 (Hall 1999). PAUP∗ (Swofford 2001) was used to calculate the uncorrected pairwise DNA sequence divergences and perform a maximum parsimony analysis with random addition and tree bisection reconnection branch swapping. All characters were unweighted and unordered. A jackknife and bootstrap analysis with 1000 replications was implemented in PAUP ∗ to assess branch support. Posterior probabilities were also assessed with 1 million Markov chain Monte Carlo generations in MRBAYES (Ronquist and Huelsbeck 2003) using the GTR + Ŵ +I model of evolution. Trees were sampled every 100 generations after the first 250 000 burn-in trees. Malapterurus electricus (Siluriformes: Malapteruridae) collected from the region was designated the outgroup. A phylogenetic analysis of Siluriformes using rag1 and rag2 nuclear genes hypothesized that Malapteruridae was the sister family to Amphiliidae (Sullivan et al. 2006). Results Amphilius rheophilus complex Geometric morphometrics The lateral analysis of 338 A. rheophilus specimens revealed some overlap in body shape among populations in the basins sampled (Figure 4). However, there is separation of specimens from the Gambie, Rio Corubal, Kaba and the syntypes of A. rheophilus from specimens collected in the Konkouré, Senegal, Niger, Loffa and Loh (St Paul) River basins. The mean body shapes of specimens from the Rio Corubal and Senegal River basins differ significantly with a Procrustes distance between the means of 0.041 (P = 0.01). The thin spline deformation reveals that specimens from the Rio Corubal have a deeper body, shorter snout, and a smaller caudal peduncle length to height ratio than Senegal River specimens. The ventral analysis of 654 A. rheophilus specimens resulted in greater overlap in body shape across the basins sampled (Figure 5). Again there was a grouping of specimens from the Gambie River, Rio Corubal, Kaba River and the syntypes. Specimens from these basins are slightly separated from specimens collected in the Journal of Natural History 531 Figure 4. Principal component scores for the lateral analysis of 338 Amphilius rheophilus. Specimens examined: Senegal (X) n = 186, Rio Corubal (△) n = 81, Konkouré (+) n = 45, Niger (o) n = 6, Loffa () n = 2, Loh (⊲) n = 3, Gambie (▽) n = 3, Kaba (⊳) n = 11, and Syntypes (p) n = 2. Bold plots represent basins where tissue samples were collected. With deformation grids (exaggerated by two). Konkouré, Senegal, Loh and Loffa rivers along principal component 1 (PC 1), and completely separated from specimens collected in the Niger River. The mean body shape of specimens collected in the Rio Corubal is significantly different from Senegal River specimens with a Procrustes distance between the means of 0.0558 (P = 0.00). The Procrustes distance between the means of Rio Corubal specimens and Niger River specimens is 0.0857 (P = 0.00). The thin spline deformation reveals that specimens from the Rio Corubal display a shorter distance from the mid-point of their branchiostegal notch to the insertion of the four mandibular barbels than specimens from the Senegal River (Figure 5). Specimens from the Niger River have an even longer distance, but are not shown here. The dorsal analysis of A. rheophilus obtained similar results, but with more overlap between basins. Standard morphometrics The principal components analysis of standard morphological measurements taken from 147 A. rheophilus specimens revealed similar results to the GM analysis 532 R.C. Schmidt and F. Pezold Figure 5. Plot of principal component scores for the ventral analysis of 654 Amphilius rheophilus. Specimens examined: Senegal (X) n = 380, Rio Corubal (△) n = 140, Konkouré (+) n = 96, Niger (o) n = 10, Loffa () n = 5, Loh (⊲) n = 3, Gambie (▽) n = 3, Kaba (⊳) n = 15, and syntypes (p) n = 2. Bold plots represent basins where tissue samples were collected. With deformation grids (exaggerated by two). (Figure 6). The first principal component primarily expressed variation in body size of specimens. Plots of other principal components generally reflect differences in body shape morphology with less influence of size. A plot of PC 2 and PC 3 shows specimens from the Rio Corubal, Loh River, Kaba River and syntypes of A. rheophilus are separated from Senegal River, Konkouré River, Loffa River, Niger River and Gambie River specimens (Figure 6). The variation observed along the second principal component is influenced by barbel length (BL), caudal peduncle length (CL) and caudal peduncle height (CH) (Table 2). The adipose fin length (AL) is responsible for most of the variation along the third principal component (Table 2), the primary axis on which the populations are separated. Univariate analyses of the log-transformed variables gave similar results and are displayed in Table 3. Journal of Natural History 533 Figure 6. Plot of PC2 to PC3 from PCA of linear measurements from 147 Amphilius rheophilus specimens. Specimens examined: Senegal (X) n = 47, Rio Corubal (△) n = 30, Konkouré (+) n = 34, Niger (o) n = 12, Loffa () n = 3, Loh (⊲) n = 6, Gambie (▽) n = 3, Kaba (⊳) n = 10, and syntypes (p) n = 2. Bold plots represent basins where tissue samples were collected. Table 2. Component loadings of the principal components analysis of 147 Amphilius rheophilus specimens. Barbel Length Standard Length Caudal Peduncle Height Caudal Peduncle Length Predorsal Length Head Length Snout Length Interorbit distance Adipose Fin Length Component 2 Component 3 −0.594 0.169 −0.345 0.343 0.022 0.096 0.225 −0.054 −0.009 −0.163 −0.004 0.116 −0.277 0.087 0.119 0.143 0.268 −0.352 Colouration Amphilius rheophilus specimens showed little variation in pigmentation across basins. All specimens showed the distinct dorsal saddles along the back that were described by n SL CL/CH As% of SL AL CL As% of HL BL IO Syntypes Gambie 2 51.9±5.6 (48.0-55.9) 1.76±0.2a (1.75-1.78) 3 29.6±2.9 (26.9-32.7) 2.4±1.1cd (2.1-27) 14.1±1.0 (13.6-14.6) 15.9±1.0abc (15.6-16.2) 47.5±1.2a (42.4-53.2) 33.4±1.2b (29.9-37.2) Rio Corubal Kaba Loh Konkouré Senegal Niger Loffa 30 34.9±7.2 (26.4-55.4) 2.0±1.1abc (1.7-2.3) 10 37.4±8.5 (20.4-47.7) 1.8±1.1ab (1.6-2.0) 6 45.9±5.3 (22.7-41.8) 2.3±1.0bcd (2.1-2.3) 34 34.7±6.3 (26.6-52.8) 2.7±1.1d (1.9-3.3) 47 36.7±7.5 (24.7-54.0) 2.6±1.1d (2.1-3.5) 12 29.8±5.3 (22.7-41.8) 2.6±1.1d (2.3-3.0) 3 41.3±7.5 (33.5-48.5) 2.6±1.1d (2.5-2.8) 16.8±1.0 (16.2-17.4) 16.5±1.1abc (15.5-17.0) 14.2±1.2 (9.5-17.8) 15.6±1.1ab (13.8-17.8) 14.9±1.1 (12.3-17.0) 15.3±1.1a (13.2-16.6) 12.9±1.1 (11.0-14.8) 16.3±1.0abc (15.1-17.4) 15.0±1.1 (12.3-17.0) 18.1±1.1c (15.1-20.1) 15.7±1.1 (12.3-17.8) 18.1±1.1c (16.6-20.0) 14.7±1.1 (11.7-17.8) 17.8±1.1bc (15.8-19.1) 17.4±1.0 (17.0-18.2) 18.2±1.0c (17.4-18.6) 56.2±1.1ab (52.5-58.9) 31.9±1.0ab (31.6-32.4) 60.4±1.1b (52.5-72.4) 33.3±1.1b (28.2-39.8) 52.5±1.1ab (44.7-63.1) 33.1±1.1b (28.9-37.2) 46.8±1.1a (42.7-52.5) 28.3±1.0a (26.9-29.5) 52.8±1.1ab (46.8-63.1) 31.5±1.1ab (28.8-34.7) 57.1±1.1ab (44.7-74.1) 31.4±1.1ab (27.5-36.3) 59.8±1.1b (51.3-64.6) 31.8±1.1ab (28.8-35.5) 57.5±1.2ab (51.3-67.6) 28.2±1.1a (26.9-30.2) 534 R.C. Schmidt and F. Pezold Table 3. Mean ± SD and (range) of standard morphometric variables for Amphilius rheophilus, groups indicated by superscript letters as assessed by Tukey’s test with P = 0.05. Journal of Natural History 535 Daget (1959). Specimens from the Niger and Senegal basins displayed slightly darker body and more distinct dorsal saddles. Kaba River specimens were slightly different in that in one location two colour morphs were present. Both of these morphs displayed the characteristic dorsal saddle pattern however, one morph was much darker overall with very distinct dorsal saddles. Gill rakers Amphilius rheophilus specimens vary in the number of gill rakers on the lower first gill arch (ceratobranchial). Specimens from the Rio Corubal and Gambie River basins have six rakers on the lower portion of the arch versus five observed on specimens from the Senegal River, Konkouré River and Niger River basins. Specimens from the Kaba River possess four or five gill rakers. Specimens from the Loh and Loffa River have six or seven rakers on the lower portion of their gill arch. The syntypes of A. rheophilus examined had six rakers on the lower arch. Cytochrome b analysis The analysis of 773 base pairs from 52 amphiliid specimens (21 A. rheophilus, 30 A. platychir and one A. atesuensis) included 222 parsimony informative characters (GenBank accession numbers GQ379942-GQ379993). The maximum parsimony analysis with jackknife/bootstrap support (Figure 7) resulted in deep, well-supported divergence within the A. platychir and A. rheophilus clades (consistency index = 0.7187, retention index = 0.9607). Amphilius rheophilus comprised three well-supported (100%) monophyletic groups representing specimens from the Konkouré River, Rio Corubal and Senegal River. The Bayesian analysis also resulted in three well-supported monophyletic A. rheophilus clades (Figure 8). Amphilius rheophilus specimens from the Rio Corubal display 9.0% divergence from the Senegal River/Konkouré River clade, and there is 4.5% divergence between specimens from the Senegal and Konkouré basins. The A. atesuensis specimen is 17.5% divergent from the A. rheophilus clade (Figure 6). Summary of Amphilius rheophilus complex The morphometric variation observed correlates with the variation observed in the number of gill rakers, and the cytochrome b data where available (Table 4). Specimens from the Konkouré and Senegal Rivers are genetically distinct from specimens from the Rio Corubal. Konkouré, Senegal and Niger River basin specimens are morphologically similar to one another and distinct from Rio Corubal and Gambie River basin specimens, including the syntypes of A. rheophilus. Specimens from the Rio Corubal and Gambie River basins are recognized here as A. rheophilus. Specimens from the Senegal River, Niger River and Konkouré River are regarded as a separate species, identified here as Amphilius aff. rheophilus sp. 1. Specimens from the Kaba River (Little Scarcies) are distinct from all other specimens in having a stout body like A. rheophilus with fewer gill rakers present (four or five); these specimens are referred to as Amphilius aff. rheophilus sp. 2. Specimens from the Loh (St Paul) and Loffa River basins have a similar morphology to those referred to as Amphilius aff. rheophilus sp. 536 R.C. Schmidt and F. Pezold Figure 7. Phylogeny inferred by cytochrome b analysis. Majority-rule consensus tree with jackknife/bootstrap support (1000 repetitions) from maximum parsimony analysis of amphiliid catfish from the Fouta Djalon with Malapterurus electricus designated as outgroup. Support < 70 represented by ∗ and the average uncorrected p-distance is shown between the major clades. Figure 8. Bayesian analysis of cytochrome b sequences under the GTR + Ŵ + I model of evolution with posterior probabilities from 750,000 post burn-in trees. 1, but they have more gill rakers on the lower arch (six or seven) and are referred to as Amphilius aff. rheophilus sp.3. Journal of Natural History 537 Table 4. Summary of morphotypes found within the Amphilius rheophilus complex with distribution. Amphilius rheophilus Amphilius aff. rheophilus sp. 1 Caudal peduncle Deeper caudal peduncle (usually under 2.0 Elongated caudal peduncle (2.0-3.5 length/height) Gill Rakers Six rakers on lower arch Four light brown dorsal saddles on darker brown body Five rakers on lower arch Four distinct light brown dorsal saddles on darker brown body Saalawol, Fetoré, Kokoulo, and Konkouré rivers (Konkouré basin); Téné and Bafing rivers (Senegal basin); Tinkisso River (Niger basin) Colouration Localities Koumba, Senta, and Finton rivers (Rio Corubal basin) and Diwe River (Gambie basin) Amphilius aff. rheophilus sp. 2 Deeper caudal peduncle (usually under 2.0 length/height) Four to five rakers on lower arch Four distinct light brown dorsal saddles on darker brown body Kaba River (Little Scarcies basin) Amphilius platychir complex Geometric morphometrics Lateral analysis of 186 A. platychir specimens revealed no distinct groupings; however, specimens from the Badi River and Fatala River have some separation from specimens collected in the Rio Corubal, Tinguilinta, Senegal, Konkouré and Niger River basins (Figure 9). Specimens from the Fatala River were significantly different from specimens collected within the Senegal River basin with a Procrustes distance between means of 0.0392 (P = 0.00). Thin spline deformation displays that Fatala River specimens have a shallower body depth, shorter adipose fin, and greater caudal peduncle length to height ratio than Senegal River basin specimens (Figure 9). Ventral analysis of 335 A. platychir specimens resulted in overlap among the various basins, but Kolenté River specimens are separated from all basins except the Fatala River (Figure 10). Fatala River specimens are significantly different from Konkouré River basin specimens with a Procrustes distance between means of 0.0422 (P = 0.00). Thin spline deformation displays Fatala River specimens with a deeper branchiostegal notch than Konkouré River basin specimens (Figure 10). Dorsal analysis of A. platychir produced similar results. Standard morphometrics The principal components analysis of measurements taken from 225 A. platychir specimens revealed less information than the GM analysis. A plot of PC 2 and PC 3 538 R.C. Schmidt and F. Pezold Figure 9. Principal component scores for 186 Amphilius platychir specimens in lateral analysis. Specimens examined: Senegal (X) n = 22, Rio Corubal (△) n = 20, Konkouré (+) n = 111, Niger (o) n = 4, A. kakrimensis (⊲) n = 1, Badi ( ) n = 2, Fatala (⊳) n = 24, and Tinguilinta (p) n = 1. Bold plots represent basins where tissue samples were collected. With deformation grids (exaggerated by two). (Figure 11) reveals a large amount of overlap between the various basins sampled. As with A. rheophilus, barbel length (BL) and caudal peduncle length (CL) contribute most to the variation seen along PC 2 (Table 5). Principal component 3 is again influenced most by adipose fin length (Table 5). The univariate analyses of the logtransformed measurements gave results similar to the GM analysis (Table 6). Notably specimens from the Fatala, Badi, Kolenté and Tinguilinta rivers have a significantly shorter adipose fin than Senegal, Rio Corubal and Konkouré River basin specimens. Colouration Colouration of A. platychir specimens is also varied across basins (Figure 12). Specimens from the Senegal, Konkouré, Rio Corubal and Gambie River basins have similar colouration. The colouration occurs, in various intensities, with specimens Journal of Natural History 539 Figure 10. Principal component scores for the ventral analysis of 335 Amphilius platychir. Specimens examined: Senegal (X) n = 46, Rio Corubal (△) n = 31, Konkouré (+) n = 179, Niger (o) n = 7, Badi ( ) n = 5, Fatala (⊳) n = 61, Kolenté ( ), and Tinguilinta (p) n = 2. Bold plots represent basins where tissue samples were collected. With deformation grids (exaggerated by two). from the Kokoulo River, Fetoré River and Saalawol River (Konkouré basin) having the most pronounced unpigmented longitudinal lines and a distinct X on their backs. This is the colouration described for A. grammatophorus (Pellegrin 1913). Specimens from the Konkouré River have varied colouration with some individuals displaying no longitudinal lines and black spots scattered across the back and sides. The paratype of A. grammatophorus inequalis was uniformly brownish but was described as having two longitudinal lines but often covered with small black dots along the back and side (Pellegrin 1935). Amphilius platychir specimens from the Fatala River and Tinguilinta River are mostly light brown to yellowish with no distinct X on their backs. They do possess faint longitudinal bands above and below the lateral line. Kolenté River, Badi 540 R.C. Schmidt and F. Pezold Figure 11. Plot of PC2 and PC3 from principal components analysis of linear measurements from 225 Amphilius platychir. Specimens examined: Senegal (X) n = 48, Rio Corubal (△) n = 21, Konkouré (+) n = 56, Badi ( ) n = 4, Fatala (⊳) n = 55, Kolenté ( ) n = 5, Tinguilinta (p) n = 6, Koba (⊳), Gambie (▽), Liberia (o) n = 9, Sierra Leone 1 (o) n = 3, Sierra Leone 2 (o) n = 3, A. platychir syntypes (o) n = 3, A kakrimensis paratype (o), A. g. inequalis syntype(o) and A. grammatophorus syntype (o). Bold plots represent basins where tissue samples were collected. Table 5. Component loadings of the principal component analysis of 225 Amphilius platychir specimens. Barbel Length Standard Length Caudal Peduncle Height Caudal Peduncle Length Predorsal Length Head Length Snout Length Interorbit distance Adipose Fin Length Component 2 Component 3 0.258 −0.044 0.092 −0.309 0.029 0.041 −0.005 0.070 −0.138 −0.183 0.081 −0.084 −0.032 0.024 0.142 0.193 0.103 −0.262 Table 6. Mean ± standard deviation and (range) of standard morphometric variables for Amphilius platychir, groups indicated by superscript as assessed by Tukey’s test with P = 0.05. A. platychir Badi 3 40.6±4.0 (36.0−43.4) 1.36±0.2ab (1.2−1.6) 4 39.2±14.8 (30.0−61.2) 1.8±1.2c (1.4−2.1) 55 35.9±7.0 (24.7−54.3) 1.7±1.1bc (1.1−2.2) 1 38.47 15.4±1.1bcd (14.1−16.3) 13.6±1.1ab (12.0−15.2) 11.8±1.1a (10.5−13.8) 15.8±1.1bc (14.5−17.4) 13.0±1.1ab (10.5−17.8) 14.8±1.1abc (11.7−17.4) 13.8 90.8 73.9±1.1abc (68.3−77.3) 75.0±1.1abc (69.2−81.3) 69.3±1.1a (50.1−91.2) 83.2 Gambie Konkouré Rio Corubal Senegal Sierra Leone 2 Sierra Leone 1 6 28.4±4.8 (23.9−35.5) 1.6±1.1abc (1.3−1.8) 1 50.4 56 40.7±9.0 (26.7−66.9) 1.61±1.1bc (1.3−2.2) 21 48.0±14.6 (30.2−78.2) 1.2±1.1a (1.0−1.6) 48 46.6±10.3 (30.7−75.2) 1.5±1.1abc (1.1−1.9) 3 62.1±2.6 (60.6−65.1) 1.63±0.0bc (1.6−1.7) 3 29.4±5.8 (23.8−25.3) 1.52±0.2abc (1.4−1.7) 9 50.6±6.3 (41.3−59.4) 1.26±0.0ab (1.2−1.3) 11.7±1.1a (10.0−13.8) 13.8±1.1abc (12.0−15.5) 18.2 16.8±1.1cd (13.7−20.0) 15.5±1.1bc (12.9−17.5) 15.7±1.1bcd (12.3−18.6) 12.9±1.1a (11.2−15.5) 16.1±1.1bcd (8.7−19.1) 14.9±1.1abc (12.6−17.4) 18.7±1.2d (15.7−20.7) 16.3±1.1c (15.6−17.5) 14.2±1.2abc (12.3−16.9) 14.7±1.0abc (14.1−15.3) 17.6±1.1cd (16.2−18.8) 15.4±1.0bc (14.6−16.4) 73.6±1.1ab (67.6−83.2) 97.7 76.7±1.1abc (64.1−90.1) 92.3±1.1cd (81.3−107.2) 90.5±1.1bcd (67.6−112.0) 88.7±1.1bcd (81.3−100.6) 82.3±1.2abc (75.3−96.7) 106.2±1.1d (90.5−119.2) A. kakrimensis A.g inequalis A. grammatophorus n SL 1 29.6 1 58.8 1 91.5 CL/CH 1.62 1.42 1.73 As% of SL AL 15.7 17.8 19.2 CL 14.7 15.3 17.2 As% of HL BL 79.1 77.5 Tinguilinta CL/CH As% of SL AL CL As% of HL BL 1.4 14.1 Kaba 1.4 13.5 Kolenté 5 40.7±11.5 (31.0−54.5) 1.5±1.1abc (1.4−1.7) 11.2±1.2a (9.3−14.1) 14.3±1.1abc (13.2−16.2) 76.6±1.1abc (72.4−81.3) Liberia Journal of Natural History 541 n SL Fatala 542 R.C. Schmidt and F. Pezold Figure 12. Colouration of Amphilius platychir specimens from various basins. (A) Fetoré River, (B) Konkouré River, (C) Kolenté River, (D) Fatala River and (E) Tinguilinta River. Scale bar is 1 cm. Journal of Natural History 543 River (lower Konkouré basin) and Kaba River (Little Scarcies) specimens are mostly brownish (darker than Fatala River and Tinguilinta River specimens) on the sides with light longitudinal banding. The syntypes of A. platychir have not retained their pigmentation because of the age of the specimens but were described as having upper parts blackish (Günther 1864). Gill rakers Specimens of A. platychir vary in the number of gill rakers observed on the lower portion of the first gill arch. Fatala River specimens possess four to six rakers on the lower arch, Kolenté River basin specimens have six, Badi River specimens have six or seven, and Tinguilinta River specimens have five or six. The lone Kaba River (Little Scarcies) specimen had six rakers on the lower arch. Senegal River basin, Konkouré River basin, Niger River basin, Gambie River basin and Rio Corubal basin specimens have seven or eight rakers on the lower arch. The syntypes of A. platychir have four to six gill rakers on the lower arch. The syntypes of A. grammatophorus, A. grammatophorus inequalis, and paratype of A. kakrimensis examined had eight rakers on the lower arch. Cytochrome b analysis The maximum parsimony analysis of 30 A. platychir specimens with jackknife/ bootstrap support (Figure 7) resulted in deep, well-supported (100%) divergence between a Fatala River clade and a clade consisting of Konkouré River, Gambie River and Senegal River basin specimens (Figure 7). The Bayesian analysis revealed a similar topology (Figure 8). The maximum parsimony analysis with average uncorrected p-distances shows the amount of divergence observed within taxa (Figure 7). Fatala River A. platychir specimens display 10.0% sequence divergence from Konkouré, Senegal, and Gambie River basin specimens. The A. platychir and A. rheophilus clades are 20.1% divergent. Summary of Amphilius platychir complex The morphometric variation correlates with variation observed in the number of gill rakers and for cytochrome b (Table 7). Amphilius platychir specimens from the Fatala and Tinguilinta Rivers, with their lighter colouration, shorter adipose fin and fewer gill rakers, are distinct from A. platychir specimens collected in the upper Konkouré River, Gambie River, Rio Corubal, Niger and Senegal River basins. The cytochrome b analysis also showed high divergence (10%) between specimens in these two groups. The specimens from Fatala and Tinguilinta Rivers also differ from the A. platychir syntypes from Sierra Leone in having a lighter colouration and shorter adipose fin (Table 7). These specimens are referred to here as Amphilius aff. platychir sp. 1. Amphilius aff. platychir sp. 1 closely matches the description (short adipose fin and light brownish colouration) of A. grammatophorus brevipinna Pellegrin 1935 described from the Kolenté River basin. Type specimens need to be examined before this is certain. The syntype we examined of A. grammatophorus inaequalis, also described from 544 R.C. Schmidt and F. Pezold Table 7. Summary of morphotypes found within the Amphilius platychir complex with distribution. Amphilius platychir Maxillary barbel Adipose fin Gill Rakers Colouration Localities Barbels less than 80% of head length Amphilius grammatophorus Barbels long, usually well over 80% of head length, may even exceed head length Fin moderately long, Fin longer, usually over 13–16% of 15% of standard standard length length 4–6 rakers on lower 7–8 rakers on lower arch arch Generally brownish, Two distinct with darker upper unpigmented lateral body lines along side with distinct X on back anterior to dorsal fin Sierra Leone, Kaba Saalawol, Fetoré, and River (Little Kokoulo rivers Scarcies), Kolenté (Konkouré basin); River, and Badi Téné and Bafing rivers River (Senegal basin); Koumba River (Rio Corubal basin); Dimma River (Gambie basin) Amphilius aff. platychir sp. 1 Barbels usually less than 80% head length Fin shorter, 10–13% of standard length 4–6 rakers on lower arch Generally light brown with faint lateral lines present Tinguilinta River and Fatala River the Friguiagbé River in the Kolenté basin, is indistinguishable from A. grammatophorus Pellegrin 1913 (eight gill rakers and long adipose fin). Interestingly, the specimens we obtained from the Kolenté basin resembled neither A. grammatophorus nor A. aff. platychir sp. 1, but were most similar to individuals taken in the Badi (lower Konkouré River basin) and Kaba basins. Specimens from these basins resemble the A. platychir syntypes in colouration and number of gill rakers and are recognized as that species. Specimens from the Gambie, Rio Corubal, Senegal and upper Konkouré River basins are also distinct from the A. platychir syntypes. These specimens have more gill rakers on the lower arch (seven or eight compared with four to six) and distinct colouration (pronounced lateral banding and X mark on the nape versus brownish with dark dorsum). Specimens from these basins are indistinguishable from A. grammatophorus Pellegrin 1913, and are recognized as such herein. The Kakrima River near the confluence with the Konkouré River is the type locality of Amphilius kakrimensis. Although the standard analysis did not show a difference between the A. kakrimensis paratype and the northern Konkouré River basin A. grammatophorus specimens, there is a difference in colouration and general morphology (Figure 12). No tissue samples were collected for specimens from the lower Kakrima Journal of Natural History 545 or Konkouré Rivers. Additional specimens and tissues from these areas within the Konkouré River basin are needed to elucidate the variation observed and resolve the status of A. kakrimensis. Discussion The analysis of morphological characters coupled with analysis of cytochrome b data indicates that there are multiple Amphilius species currently recognized as A. platychir and A. rheophilus in Guinea. At least four species have been identified within the Amphilius rheophilus complex. Although A. rheophilus specimens from the Rio Corubal and Gambie basins differed in the standard morphological analysis, this may be a result of the small sample size (three) and small size of the specimens (27–33 mm SL) from the Gambie. The four species of this group appear to be allopatric because none were taken in the same stream. None of the A. rheophilus complex species were collected in the Fatala, Tinguilinta or Kolenté River basins. As with species of the A. rheophilus complex, species of the A. platychir complex are largely allopatric across the basins sampled, except for two instances. One specimen of Amphilius aff. platychir sp. 1 was collected in the upper Rio Corubal basin with A. grammatophorus, and a single specimen of A. grammatophorus was taken with Amphilius aff. platychir sp. 1 near the mouth of the Fatala River. The distribution patterns of fish species native to the Fouta Djalon region was described by Daget (1962). Atlantic-Guinean taxa are distributed within the western basins of the Fouta Djalon (Gambie River, Fatala River, Konkouré River, Rio Corubal, Tinguilinta River and Kaba River) that are characterized by relatively wet climate and forest-lined stream banks. Sudano-Guinean taxa are distributed within basins that originate in the Fouta Djalon and flow east (Niger and Senegal Rivers) through savanna lands that periodically flood. Daget also recognized an amphiGuinean fish fauna occurring in the upper reaches of the Atlantic-Guinean and Sudano-Guinean streams. He ascribed the Amphilius species from the Fouta Djalon to the amphi-Guinean group (Daget 1962). Distributions of the species forming the A. platychir and A. rheophilus complexes mirror the distribution patterns of other species endemic to the Fouta Djalon region. A new species of killifish (Epiplatys guineensis) was described from the middle Konkouré River, whereas a closely related species (Epiplatys fasciolatus) is found in other coastal rivers (Fatala River, Badi River and Kolenté River; Ramond 1994). In describing several new species of bariliin cyprinid fish, Howes and Teugels (1989) described Raiamas levequei from the middle Konkouré River basin, with Raiamas senegalensis being distributed throughout the Senegal and Niger River basins. Leptocypris konkoureensis was also described from the middle Konkouré River with Leptocypris guineensis found in the Niger River basin (Howes and Teugels 1989). A large number of endemic fish species is found within the western basins of the Fouta Djalon (Daget 1962; Hugueny 1989). High levels of endemism are also observed in freshwater zooplankton and molluscs from the area (Brown 1980; Dumont 1981). These fauna also follow the Sudano-Guinean and Atlantic-Guinean distribution pattern (Brown 1980; Dumont 1981). A study of dactylogyrids parasitizing cyprinids from Guinea also revealed many endemic species exhibiting this same distribution pattern (Guegan and Lambert 1990, 1991). 546 R.C. Schmidt and F. Pezold The Fouta Djalon highlands have retained forest galleries for millions of years (Hugueny and Lévêque 1994). Hugueny and Lévêque (1994) proposed that the faunistic divergence between the Sudano-Guinean and Atlantic-Guinean species could be the result of three factors: the upsurge of geographical barriers that separated the two regions; differing ecological conditions (Sudano-Guinean being more open savannah versus forested and mountainous streams in the western areas); and the presence of refuges within western basins which allowed the Atlantic-Guinean fauna to persist during climatic fluctuations (Mayr and Hara 1986). Additional species within the West African amphiliid complex were discovered through analysis of both morphological and molecular characters. The high amount of genetic diversity compared with the overall similarity in morphology is quite stunning. The high degree of genetic divergence among samples indicates significant periods of separation of Atlantic- and Sudano-Guinean populations of both species complexes, and some isolation within each complex for populations in particular coastal streams. Interestingly we see more genetic divergence between the Konkouré and Senegal River basins within the A. rheophilus complex than observed within the A. platychir complex. This would suggest differing rates of evolution between the complexes or the possibility that vicariant events affected the complexes differently. The authors are currently examining additional amphi-Guinean taxa and nuclear markers to better understand the phylogeography of the area. The relatively staid morphology suggests strong selective pressure for the streamadapted benthic body form. Differences in pigmentation appear related to different benthic habits in the complexes. Members of the A. platychir complex were typically found in eelgrass beds associated with the blades of grasses, compared with the gravel riffles and races inhabited by A. rheophilus. Species in the A. platychir complex are characterized by camouflaging horizontal bands on the flanks, whereas members of the A. rheophilus complex are barred across the dorsum and resemble the gravel streambed from above. It is probable that molecular analyses of Amphilius populations across their range will reveal the presence of additional cryptic species. A complete systematic review of the amphiliid species from Guinea and surrounding areas is currently in preparation. Acknowledgements Funding was provided from the Critical Ecosystem Partnership Fund administered by Conservation International, the HHMI/ULM Undergraduate Science Education Program and by NSF-OISE 0080699 to FP. We thank M. Elimane Diop, Baidi Samoura, Bangaly Kaba, Mamadou Camara, Brook Fluker, Stephanie McCormick, Laura George, Leslie Patterson, Jon Anderson, Scott Large, Fabio Moretzsohn, Morgan Kilgour, and members of the 2002–2003 ULM Guinea expeditions for invaluable assistance in the field and laboratory. Samba Diallo and Bakary Coulibaly (deceased) assisted with permits and logistical support for the fieldwork. We would also like to thank the following institutions and their support staff for graciously lending type and additional specimens: Melanie L.J. Stiassny and Barbara Brown American Museum of Natural History, New York (AMNH), Patrick Campbell Natural History Museum, London (BMNH), Patrice Pruvost and Romain Causse Muséum nationale d’Historie naturelle, Paris (MNHN), Emmanuel Vreven and Migüel Parrent Royal Museum for Central Africa, Tervuren, Belgium (MRAC) and Jeff T. Williams Smithsonian Institution National Museum of Natural History, Washington (USNM). Alfred Thomson and Mike Hardman both provided insightful reviews that greatly improved this manuscript. Journal of Natural History 547 References Barluenga M, Stölting KN, Salzburger W, Muschick M, Meyer A. 2006. Sympatric speciation in Nicaraguan crater lake cichlid fish. Nature 439:719–723. Bookstein FL. 1991. Morphometric tools for landmark data. Cambridge: Cambridge University Press; 435 pp. Boulenger GA. 1898. On the habit of the Siluroid fish Anoplopterus platychir, Gthr. Ann Mag Nat Hist. 8(7):447–448. Boulenger GA. 1911. Catalogue of fresh-water fishes of Africa in the British Museum (Natural History) London: Trustees of British Museum (Natural History). Brown DS. 1980. Freshwater snails of Africa and their medical importance. London: Taylor & Francis. Daget J. 1959. Description de trio poissons nouveaux de la Gambie: Barbus niokoloensis n. sp., Chrysichthys johnelsi n. sp. et Amphilius rheophilius n. sp. Bull Inst Franç Afrique Noire. 21:689–694. Daget J. 1962. Les poisons du Fouta Djalon et de la Basse Guinee. Mem Inst Franç Afrique Noire. 65:1–210. Daget J. 1963. La Réserve naturelle intégrale du Mont Nimba. XXVII.—Poissons (deuxième note). Mém Inst Franç Afrique Noire. 66:573–600. Dumont HJ. 1981. Cladocera and free-living Copepoda from the Fouta Djalon and adjacent mountain areas in West Africa. Hydrobiologia 85:97–116. Egge JJD, Simmons AM. 2006. The challenge of truly cryptic diversity: diagnosis and description of a new madtom catfish (Ictaluridae: Noturus). Zool Scripta. 35(6):581–595. Goodall CR. 1991. Procrustes methods in the statistical analysis of shape. J Roy Stat Soc. Ser B. 53(2):285–339. Guegan JF, Lambert A. 1990. Twelve new species of dactylogyrids (Platyhelminthes, Monogenea) from West African barbels (Telostei, Cyprinidae), with some biogeographical implications. Syst Parasitol. 17:153–181. Guegan JF, Lambert A. 1991. Dactylogyrids (Platyhelminthes: Mongenea) of Labeo (Teleostei: Cyprinidae) from West African coastal rivers. J Helminthol Soc Wash. 58(1):85–99. Guill JM, Hood CS, Heins DC. 2003. Body shape variation within and among three species of darters (Perciformes: Percidae). Ecol Freshwater Fish. 12:134–140. Günther A.1864. Catalogue of the fishes in the British Museum, vol. 5.–Catalogue of the physostomi, containing the families Siluridae, Characinidae, Haplochitonidae, Sternoptychidae, Scopelidae, Stomiatidae in the collection of the British Museum. London: Trustees of British Museum (Natural History). Gunther A. 1902. Last account of fishes collected by Mr. R. B. N. Walker, C. M. Z. S., on the Gold Coast. Proc Zool Soc Lond. 2(2):330–339. Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser. 41:95–98. Hankison SJ, Childress MJ, Schmitter-Soto JJ, Ptacek MB. 2006. Morphological divergence within and between the Mexican sailfin mollies, Poecilia velifera and Poecilia petenensis. J Fish Biol. 68:1610–1630. Howes GJ, Teugels GG. 1989. New bariliin cyprinid from West Africa, with a consideration of their biogeography. J Nat Hist. 23:873–902. Hugueny B. 1989. West African rivers as biogeographic islands: species richness of fish communities. Oecologia 79:236–243. Hugueny B, Lévêque C. 1994. Freshwater fish zoogeography in West Africa: faunal similarities between river basins. Environ Biol. 39:365–380. Humphries CJ. 2002. Morphology, shape and phylogeny. 1st ed. New York (NY): Taylor Francis. Chapter 2, Homology, characters and continuous variables; p. 8–26. Leviton AE, Gibbs RH, Heal E, Dawson CE. 1985. Standards in ichthyology and herpetology. 548 R.C. Schmidt and F. Pezold Part 1. Standard symbolic codes for institutional resources collections in ichthyology and herpetology. Copeia 1985:802–832. Mayr E, O’Hara RJ. 1986. The biogeographic evidence supporting the Pleistocene forest refuge hypothesis. Evolution 40(1):55–67. Murphy WJ, Collier GE. 1996. A molecular phylogeny for aplocheiloid fishes (Atherinomorpha, Cyprinodontiformes): the role of vicariance and the origins of annualism. Mol Biol Evol. 14(8):790–799. Near TJ, Porterfield JC, Page LM. 2000. Evolution of cytochrome b and the molecular systematics of Ammocrypta (Percidae: Etheostomatinae). Copeia 2000(3):701–711. Palumbi SR. 1996. Molecular systematics. 2nd ed. Sunderland (MA): Sinauer Associates, Inc. Chapter 7, Nucleic acids II: The polymerase chain reaction; pp. 205–247. Parsons KJ, Robinson BW, Hrbek T. 2003. Getting into shape: an empirical comparison of traditional truss-based morphometric methods with a newer geometric applied to New World cichlids. Environ Biol Fishes. 67:417–431. Paugy D, Leveque C, Teugels GG. 2003. The fresh and brackish water fishes of West Africa. 2nd ed. IRD Editions. Paris: Publications Scientifiques du Museum, MRAC Pellegrin J.1913. Poissons nouveaux de Guinee Francaise recueillis par M. Pobeguin. Bull Soc Zool France. 38:236–241. Pellegrin J. 1935. Poissons de Guinée française recueillis par M. waterlot description d’une espèce et de deux variétés nouvelles. Bull Soc Zool France. 60:462–466. Poche F. 1902. Uber das vorkommen von Anoplopterus platychir (Gthr) in Westafrika. Zool Anz. 25:121–122. Ramond R. 1994. Epiplatys guineensis, a new species of killifish from Guinea, West Africa (Pisces: Cyprinodontidae). Ichthyol Expl Freshwaters. 5(4):365–370. Reis RE, Trajano E, Hingst-Zaher E.2006. Shape variation in surface and cave populations of the armoured catfishes Ancistrus (Siluriformes: Loricariidae) from the São Domingos karst area, upper Tocantins River, Brazil. Environ Biol Fishes. 68:414–429. Roberts, TR.1975. Geographical distributions of African freshwater fishes. Zool J Linn Soc. 249–319. Rohlf FJ. 2004. TpsDig ver 1.40. Stony Brook, (NY): Department of Ecology and Evolution, State University of New York. Rohlf FJ, Slice DE. 1990. Extensions of the procrustes method for the optimal superimposition of landmarks. Syst Zool. 39:40–59. Ronquist F, Huelsenbeck JP. 2003. MRBAYES 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19:1572–1574. Skelton PH. 1981. The description and osteology of a new species of Gephyroglanis (Siluriformes, Bagridae) from the Olifants River, South West Cape, South Africa. Ann Cape Prov Mus Nat Hist. 13:217–249. Skelton PH. 1984. A systematic revision of species of the catfish genus Amphilius (Siluroidei, Amphiliidae) from east and southern Africa. Ann Cape Prov Mus. 16(3):41–71. Sullivan JP, Lundberg JG, Hardman M. 2006. A phylogenetic analysis of the major groups of catfishes (Teleostei: Siluriformes) using rag1 and rag2 nuclear gene sequences. Mol Phylogen Evol. 41(3):636–662. Swofford DL. 2001. PAUP∗ : Phylogenetic Analysis Using Parsimony (∗ and other methods.) Version 4. Sunderland (MA): Sinauer Associates. Teugels, GG, Skelton PH, Lévêque C. 1987. A new species of Amphilius (Pisces, Amphiliidae (Pisces, Amphiliidae) from the Konkouré basin, Guinea, West Africa. Cybium 11(1): 93–101. Thompson JD, Higgins DG, Gibson TJ. 1994. ClustalW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22:4673–4680. Journal of Natural History 549 Valentin A, Sévigny J-M, Chanut J-P. 2002. Geometric morphometrics reveals body shape differences between sympatric redfish Sebastes mentalla, Sebastes fasciatus and their hybrids in the Gulf of St Lawrence. J Fish Biol. 60:857–875. Vreven EJ, Teugels GG. 2005. Redescription of Mastacembelus liberiensis Boulenger, 1898 and description of a new West African spiny-eel (Synbranchiformes: Mastacembelidae) from the Konkouré River basin, Guinea. J Fish Biol. 67:332–369. Appendix 1 Materials examined listed by country and river basin (Catalogue number, prefecture, river, locality, latitude, longitude, collector, and standard length) Amphilius atesuensis Ghana: BMNH 1903.4.24.74-77: Atesu River, Coll. R. B. N. Walker, (syntypes of A. atesuensis). Liberia: AMNH 12313: Stream near Kaleata, Coll. G. W. Harley, 43.8 mm (holotype of A. pictus). Guinea: St Paul River basin: AMNH 248702: Nzérékoré, Loh River, Gnampara, 7◦ 54.747 N, 8◦ 55.458 W, Coll. F. Pezold et al.; CNSHB Uncat: Nzérékoré, Loh River, Gbegnema, 8◦ 72.022 N, 8◦ 43.675 W, Coll. F. Pezold et al., 36.1–38.4 mm. Amphilius kakrimensis Guinea: Konkouré River basin: MNHN 1986-0600: Kakrima River, Kasseri, 10◦ 16.0 N, 12◦ 28.0 W, Coll. C. Leveque, 29.6 mm (paratype of A. kakrimensis). Amphilius platychir Guinea: Tinguilinta River basin: ABS-11: Boké, Kewewol River, Campement, 11◦ 22 N, 13◦ 58 W, Coll. Samoura and others, 24.3–33.7 mm; FP03-13: Boke, Wonkoun River, Korira, 10◦ 40.286 N, 14◦ 15.623 W, Coll. F. Pezold et al., 26.7 mm; FP03-21: Boke, Tinguilinta River, Kewewol, 11◦ 02.066 N, 13◦ 58.378 W, Coll. F. Pezold et al., 23.9–35.4 mm. Small coastal basins: ABS-05: Boffa, Dangara River, Kissinssing, 10◦ 17 N, 14◦ 11 W, Coll. Samoura and others, 25.8– 53.2 mm; ABS-07: Boffa, Kissiling River, Kissiling, 10◦ 22 N, 14◦ 18 W, Coll. Samoura and others, 44.4 mm; AMNH 248158: Boké, Tamanransi River, Korera, Coll. Samoura and others, 43.8 mm;. Fatala River basin: ABS-01: Boffa, Koumbouya River, bridge at Bakoro, 10◦ 18 N, 14◦ 08, Coll. Samoura and others, 25.9–57.0 mm; ABS-02: Boffa, Lébé River, Segebadé, 10◦ 19 N, 14◦ 02 W, Coll. Samoura and others, 26.4–42.8 mm; FP03-02: Boffa, Sanyan River, Foutan near bridge, 10◦ 17.512 N, 14◦ 03.226 W, Coll. F. Pezold et al., 31.9–54.2 mm; FP03-05: Boffa, Koumbouya River, Bakoro, 10◦ 18.467 N, 14◦ 01.271 W, Coll. F. Pezold et al., 32.4–45.8 mm; FP03-08: Boffa, Koumbouya River, Fatala estuary-Toukeren, 10◦ 14.952 N, 14◦ 00.14 W, Coll. F. Pezold et al., 25.9–35.9 mm. Rio Corubal basin: ABS-32: Mali, N’bagou River, N’bagou, 11◦ 42 N, 12◦ 20 W, Coll. Samoura and others, 29.2–41.0 mm; AMNH 248676: Labé, Koumba River, at bridge way Labé-Mali, 11◦ 39 N, 12◦ 17 W, Coll. Samoura and others, 40.4–69.7 mm; AMNH 248677: Labé, Koumba River, Koumba, 11◦ 39 N, 12◦ 17 W, Coll. F. Pezold et al., 30.4– 74.8 mm; AMNH 248678: Mali, Koumba River, N’bagou, 11◦ 42 N, 12◦ 20 W, Coll. F. Pezold et al., 28.4–68.6 mm; CNSHB Uncat: Labé, Koumba River, Dalen, 11◦ 30 N, 12◦ 14 W, Coll. Samoura and others, 38.0 mm. Kolenté River basin: AMNH 248674: Kindia, Gbassa River, Friguiagbé, 09◦ 56 N, 12◦ 57 W, Coll. Samoura and others, 37.8–60.9 mm; FP03-94: Kinda, Senta River, Segueya, 10◦ 00.192 N, 12◦ 46.834 W, Coll. F. Pezold et al., 31.0–39.3 mm; FP0396: Kindia, Kilissi River, Senta, 09◦ 56.935 N, 12◦ 52.982 W, 51.5–54.9 mm; MNHN 1935-0191: Friguiagbé River, near Kindia, Coll. M. Waterlot, 58.8 mm (syntype of A. grammatophorus inequalis). Konkouré River basin: ABS-41: Mamou, Konkouré River, Lissan, 10◦ 17 N, 12◦ 24 W, Coll. Samoura and others, 49.3 mm; AMNH 248159: Labé, Sala River, Landou, 11◦ 19 N, 550 R.C. Schmidt and F. Pezold 12◦ 35 W, Coll. Samoura and others, 31.6–45.6 mm; AMNH 248161: Labe, Garambewol River, Dara, 11◦ 15 N, 12◦ 21 W, Coll. Samoura and others; AMNH 248679: Labé, Banewa River, Karere, 11◦ 32.349 N, 12◦ 21.792 W, Coll. F. Pezold et al., 42.8–43.2 mm; AMNH 248680: Labé, Saalawol River, N’Niogou, 11◦ 18.004 N, 12◦ 26.580 W, Coll. F. Pezold et al., 26.0– 39.1 mm; AMNH 248681: Pita, Kokoulo River, Kokoulo, 11◦ 06 N, 12◦ 23 W, Coll. F. Pezold et al.; AMNH 248682: Pita, Kokoulo River, Sintali, 11◦ 05.302 N, 12◦ 25.074 W, Coll. F. Pezold et al., 25.4–37.8 mm; CNSHB Uncat: Labé, Saalawol River, Nyogou, 11◦ 18 N, 12◦ 26 W, Coll. Samoura and others, 31.1–39.9 mm; CNSHB Uncat: Pita, Mita River, Massi, 10◦ 55 N, 12◦ 23 W, Coll. Samoura and others, 34.5–47.3 mm; CNSHB Uncat: Pita, Saalawol River, Lannou, 11◦ 19.716 N, 12◦ 35.734 W, Coll. F. Pezold et al., 26.9–35.3 mm; CNSHB Uncat: Pita, Fetoré River, Wassa(tinbitounni), 11◦ 05 N, 12◦ 33 W, Coll F. Pezold et al.; CNSHB Uncat: Pita, Kokoulo River, Kokoulo, 11◦ 06 N, 12◦ 23 W, Coll. F. Pezold et al.; CNSHB Uncat: Labé, Gorore River, Diari, 11◦ 19 N, 12◦ 29 W, Coll. Samoura and others, 29.7 mm; CNSHB Uncat: Dalaba, Badi River, Mafara, 10◦ 54 N, 11◦ 58 W, Samoura and others, 37.7–49.3 mm; CNSHB Uncat: Labé, Garambewol River, Dara, 11◦ 15.324 N, 12◦ 21.653 W, Coll. F. Pezold et al., 26.9– 35.2 mm; FP-03-71: Mamou, Konkouré River, Sangoya - Dandemiti Bridge, 10◦ 29.521 N, 12◦ 22.121 W, Coll. F. Pezold et al., 37.3–65.0 mm; FP03-72: Kindia, Konkouré River, Konkouré, 10◦ 25.218 N, 12◦ 24.014 W, Coll. F. Pezold et al., 35.1–53.0 mm; FP03-73: Kindia, Kombira River, Louguin, 10◦ 21.030 N, 12◦ 26.459 W, Coll. F. Pezold et al., 30.4–55.9 mm; FP03-75: Kindia, Konkouré River, Goubhi (Road Mamou-Kindia), 10◦ 17.365 N, 12◦ 24.736 W, Coll. F. Pezold et al., 42.5–42.8 mm; MNHN 2009-094: Pita, Fetoré River, Wassa(tinbitounni), 11◦ 05 N, 12◦ 33 W, Coll. F. Pezold et al.; FP03-102: Kindia, Gastonya River, Kamaraboudji, 09◦ 54.018 N, 13◦ 01.334 W, Coll. F. Pezold et al., 30.0–34.9 mm; FP03-103: Kindia, Badi River, Koliboya, Coll. F. Pezold et al., 25.2 mm; MNHN 1913-0231: Kokoulo River, Coll. M. Pobéguin, 91.5 mm (syntype of A. grammatophorus); Little Scarcies basin: FP03-70: Mamou, Tiguinbeli River, Kansa(Oure-Kaba), 10◦ 08.891 N, 11◦ 33.765 W, Coll. F. Pezold et al., 37.8 mm. Gambie River basin: AMNH 248160: Tougué, Kioma River, Teliko, 11◦ 20 N, 11◦ 51 W, Coll. Samoura and others, 32.0–35.0 mm; AMNH 248675: Tougué, Diwe River, Sannou, 11◦ 27 N, 12◦ 03 W, Coll. Samoura and others, 49.2 mm. Senegal River basin: AMNH 248673: Dalaba, Siragoure River, Kourou, 10◦ 52 N, 12◦ 04 W, Coll. Samoura and others, 39.1–58.1 mm; AMNH 248683: Dalaba, Téné River, Thianguelnoussi, 10◦ 48.890 N, 12◦ 14.273 W, Coll. F. Pezold et al., 31.0–65.7 mm; AMNH 248684: Mamou, Bafing River, Sogotoro, 10◦ 39.745 N, 11◦ 45.102 W, Coll. F. Pezold et al., 28.1–57.7 mm; AMNH 248685: Mamou, Bafing River, Basambaya, 10◦ 30.746 N, 11◦ 59.273 W, Coll. F. Pezold et al., 41.40–60.9 mm; AMNH 248710: Dalaba, Téné River, Thianguelnoussi, 10◦ 48 N, 12◦ 14 W, Coll. Samoura and others, 31.3–72.2 mm; CNSHB Uncat: Dalaba, Téné River, Kébali, 10◦ 56 N, 12◦ 10 W, Coll. Samoura and others; CNSHB Uncat: Mamou, Bafing River, Basambaya, 10◦ 30 N, 11◦ 59 W, Coll. Samoura and others, 40.7–53.3 mm; MNHN 2009-095: Dalaba, Bodiwol River, Mafara, 10◦ 54.054 N, 11◦ 58.740 W, Coll. F. Pezold et al., 38.2–53.68 mm; MRAC 2009-05-P-16-35: Pita, Kokoulo River, Sintali Bridge, 11◦ 05 N, 12◦ 25 W, Coll. Samoura and others, 25.0–41.0 mm; MRAC 2009-05-P-1-15: Dalaba, Mita River, Fougou, 10◦ 55.487 N, 12◦ 23.609 W, Coll. F. Pezold et al., 35.8–49.6 mm; USNM 395062: Pita, Fetoré River, Wassa (tinbitounni), 11◦ 05 N, 12◦ 33 W, Coll. F. Pezold et al. Niger River basin: CNSHB Uncat: Dalaba, Siragoure River, Kourou Maninka, 10◦ 52.852 N, 11◦ 04.514 W, Coll. F. Pezold et al., 34.0–52.0 mm. Sierra Leone: Small Coastal basins: BMNH 1949.11.1.1: Freetown, 35.3 mm; BMNH 1976.11.12.88-90: Regents stream, 23.8–28.4 mm. Sewa River basin: BMNH 1932.5.18.50-53: Bagbwe River, near Tembutinda, Coll. G. Bates, 60.5–65.1 mm; Unknown river basins: BMNH 2001.12.5.574-577: 36.0–43.4 mm (syntypes of A. platychir). Liberia: Unknown river basins: MRAC 73-27-P-905-911: Bong County, Wene Town, Coll. W. G. Johnson, 41.3–55.1 mm; MRAC 73-27-P-912: Bong County, Wene Town, W. G. Johnson, 59.4 mm; MRAC 73-27-P-919-922: Bong County, Konoyee, Coll. W. G. Johnson, 44.8–58.2 mm. Journal of Natural History 551 Amphilius rheophilus Guinea: Rio Corubal basin: ABS-32: Mali, N’bagou River, N’bagou, 11◦ 42 N, 12◦ 20 W, Coll. Samoura and others; AMNH 248703: Gaoual, Senta River, Bridge at Hakkoude Thiangui, 11◦ 39 N, 13◦ 43 W, Coll. Samoura and others, 23.3–41.6 mm; AMNH 248704: Gaoual, Koumba River, Keneri, 11◦ 39 N, 13◦ 01 W, Coll. Samoura and others, 36.1–37.2 mm; AMNH 248706: Labé, Koumba River, Bridgeway Labé-Mali, 11◦ 39 N, 12◦ 17 W, Coll. Samoura and others; AMNH 248708: Gaoual, Finton River, Bridge at Dandoun (Kembra), 11◦ 38.450 N, 13◦ 17.094 W, Coll. F. Pezold et al., 21.2–36.6 mm; AMNH 248709: Labé, Banewa River, Karere, 11◦ 32.349 N, 12◦ 21.792 W, Coll. F. Pezold et al.; AMNH 248711: Gaoual, Senta River, Senta Bridge, 11◦ 45.515 N, 13◦ 33.539 W, Coll. F. Pezold et al.; CNSHB Uncat: Gaoual, Senta River, Senta Bridge, 11◦ 45.515 N, 13◦ 33.539 W, Coll. F. Pezold et al.; CNSHB Uncat: Mali, Koumba River, N’bagou, 11◦ 42 N, 12◦ 20 W, Coll. F. Pezold et al., 22.8–39.8 mm; CNSHB Uncat: Labé, Banewa River, Karere, 11◦ 32.349 N, 12◦ 21.792 W, Coll. F. Pezold et al.; CNSHB Uncat: Gaoual, Bantala River, Bantala, 11◦ 43 N, 12◦ 53 W, Coll. Samoura and others, 32.3 mm; MNHN 2009-93: Labé, Banewa River, Karere, 11◦ 32.349 N, 12◦ 21.792 W, Coll. F. Pezold et al.; MRAC 2009-05-P-36-52: Labé, Koumba River, Koumba, 11◦ 39 N, 12◦ 17 W, Coll. Samoura and others; USNM 395061: Labé, Koumba River, Bridgeway Labé-Mali, 11◦ 39 N, 12◦ 17 W, Coll. Samoura and others. Konkouré River basin: AMNH 248686: Labé, Sala River, Landou, 11◦ 19 N, 12◦ 35 W, Coll. Samoura and others, 25.0–42.5 mm; AMNH 248687: Labé, Garambewol River, Dara, 11◦ 15 N, 12◦ 21 W, Coll. Samoura and others; AMNH 248688: Pita, Kokoulo River, Sintali Bridge, 11◦ 05 N, 12◦ 25 W, Coll. Samoura and others, 19.0–37.7 mm; AMNH 248693: Labe, Garambewol River, Dara, 11◦ 15.324 N, 12◦ 21.653 W, Coll. F. Pezold et al., 37.69 mm; AMNH 248696: Dalaba, Miti River, Fougou, 10◦ 55.487 N, 12◦ 23.609 W, Coll. F. Pezold et al.; AMNH 248698: Mamou, Konkouré River, Sangoya-Dandemiti Bridge, 10◦ 29.521 N, 12◦ 22.121 W, Coll. F. Pezold et al., 28.0–52.3 mm; AMNH 248699: Kindia, Kombira River, Louguin, 10◦ 21.030 N, 12◦ 26.459 W, Coll. F. Pezold et al., 29.7–35.9 mm; CNSHB Uncat: Pita, Mita River, Massi, 10◦ 55 N, 12◦ 23 W, Coll. Samoura and others, 25.6– 43.9 mm; CNSHB Uncat: Pita, Saalawol River, Lannou, 11◦ 19.716 N, 12◦ 35.734 W, Coll. F. Pezold et al., 29.8–30.6 mm; CNSHB Uncat: Dalaba, Mita River, Fougou, 10◦ 55.487 N, 12◦ 23.609 W, Coll. F. Pezold et al.; CNSHB Uncat: Kindia, Konkouré River, Konkouré Bridge, 10◦ 25.218 N, 12◦ 24.014 W, Coll. F. Pezold et al., 27.3–36.3 mm; FP03-75: Kindia, Konkouré River, Goubiti (Road Mamou-Kindia), 10◦ 17.365 N, 12◦ 24.736 W, Coll. F. Pezold et al., 30.4– 36.9 mm; MRAC 2009-05-P-53-64: Dalaba, Mita River, Fougou, 10◦ 55.487 N, 12◦ 23.609 W, Coll. F. Pezold et al. Little Scarcies basin: ABS-43: Mamou, Tiguiberi River, Kansa (Afflutent Penselli), 10◦ 08 N, 11◦ 33 W, Coll. Samoura and others, 33.5–39.4 mm; ABS-45: Mamou, Kaba River, Kouloundala, 10◦ 10 N, 11◦ 49 W, Coll. Samoura and others, 25.7–49.6 mm; FP03-68: Mamou, Pinselli River, Oure Kaba, 10◦ 09.334 N, 11◦ 40.048 W, Coll. F. Pezold et al., 36.4 mm; FP03-69: Mamou, Kaba River, Berteya, 10◦ 10.584 N, 11&ring;49.407 W, Coll. F. Pezold et al., 31.0–49.3 mm; FP03-70: Mamou, Tiguiberi River, Kansa (Oure-Kaba), 10◦ 08.891 N, 11◦ 33.765 W, Coll. F. Pezold et al., 26.8–33.7 mm; Gambie River basin: AMNH 248705: Tougue, Diwe River, Sannou, 11◦ 27 N, 12◦ 03 W, Coll. Samoura and others, 23.8–33.0 mm. Senegal River basin: ABS-26: Labé, Donbele River, Dionfo, 11◦ 19 N, 12◦ 00 W, Coll. Samoura and others; ABS-27: Tougué, Kioma River, Teliko, 11◦ 20 N, 11◦ 51 W, Coll. Samoura and others, 23.8–48.0 mm; AMNH 248689: Dalaba, Siragoure River, Kourou, 10◦ 52 N, 12◦ 04 W, Coll. Samoura and others, 23.5–37.9 mm; AMNH 248690: Dalaba, Badi River, Mafara, 10◦ 54 N, 11◦ 58 W, Coll. Samoura and others; AMNH 248691: Mamou, Bafing River, Basambaya, 10◦ 30 N, 11◦ 59 W, Coll. Samoura and others; AMNH 248692: Mamou, Bafing River, Kegneko, 10◦ 32 N, 11◦ 47 W, Coll. Samoura and others, 44.3–48.7 mm; AMNH 248694: Dalaba, Ditiwol River, Ditin, 10◦ 52.647 N, 12◦ 10.950 W, Coll. F. Pezold et al., 23.5–33.6 mm; AMNH 248695: Dalaba, Téné River, Thianguelnoussi, 10◦ 48.890 N, 12◦ 14.273 W, Coll. F. Pezold et al., 21.1–63.0 mm; AMNH 248697: Mamou, Poukouwol River, Timbo-Niagara, 10◦ 42.576 N, 11◦ 53.044 W, Coll. F. Pezold et al., 25.3–37.4 mm; AMNH 248701: Mamou, Bafing River, 552 R.C. Schmidt and F. Pezold Sogotoro, 10◦ 39.745 N, 11◦ 45.102 W, Coll. F. Pezold et al.; AMNH 248707: Dalaba, Téné River, Kebali, 10◦ 56 N, 12◦ 10 W, Coll. Samoura and others; CNSHB Uncat: Dalaba, Téné River, Thianguelnoussi, 10◦ 48 N, 12◦ 14 W, Coll. Samoura and others; CNSHB Uncat: Dalaba, Badi River, Mafara, 10◦ 54 N, 11◦ 58 W, Coll. Samoura and others; CNSHB Uncat: Dalaba, Téné River, Kebali, 10◦ 56 N, 12◦ 10 W, Coll. Samoura and others; CNSHB Uncat: Mamou, Bafing River, Basambaya, 10◦ 30 N, 11◦ 59 W, Coll. Samoura and others; CNSHB Uncat: Mamou, Bafing River, Sogotoro, 10◦ 39.745 N, 11◦ 45.102 W, Coll. F. Pezold et al.; CNSHB Uncat: Mamou, Bafing River, Kegneko, 10◦ 32.902 N, 11◦ 47.319 W, Coll. F. Pezold et al., 33.2–35.0 mm; CNSHB Uncat: Dabola, Djendjou River, Kodala (Mamou-Dabola), 10◦ 43 N, 11◦ 27 W, Coll. Samoura and others, 29.7–43.9 mm; FP03-55: Dalaba, Kassagui River, Lahoya, 11◦ 11 N, 12◦ 03 W, Coll. F. Pezold et al., 23.6–32.1 mm; FP03-81: Mamou, Bafing River, Basambaya, 10◦ 30.746 N, 11◦ 59.273 W, Coll. F. Pezold et al., 28.0–39.1 mm; FP03-90: Dabola, Djedjou River, Kodala, 10◦ 43.811 N, 11◦ 27.816 W, Coll. F. Pezold et al., 24.7–47.1 mm; MNHN 2009092: Dalaba, Badi River, Mafara, 10◦ 54 N, 11◦ 58 W, Coll. Samoura and others; USNM 395060: Dalaba, Badi River, Mafara, 10◦ 54 N, 11◦ 58 W, Coll. Samoura and others. Niger River basin: AMNH 248700: Dabola, Tinkisso River, Dam, 10◦ 43.676 N, 11◦ 10.113 W, Coll. F. Pezold et al., 33.5 mm; CNSHB Uncat: Dabola, Tinkisso River, Bissikrima, 10◦ 50.489 N, 10◦ 55.656 W, Coll. F. Pezold et al., 40.0 mm; CNSHB Uncat: Dalaba, Siragoure River, Kourou maninka, 10◦ 52.852 N, 11◦ 04.514 W, Coll. F. Pezold et al., 22.5–40.9 mm. Loffa River basin: FP03132: Macenta, Loffa River, Bongamadou, 8◦ 29.223 N, 9◦ 27.051 W, Coll. F. Pezold et al. St. Paul River basin: ; FP03-125: Nzérékoré, Loh River, Gbegnema, 8◦ 72.022 N, 8◦ 43.675 W, Coll. F. Pezold et al.; FP03-126: Nzérékoré, Loh River, Gnampara, 7◦ 54.747 N, 8◦ 55.458 W, Coll. F. Pezold et al. Senegal: MNHN 1959-0147: Gambie River, Mpantie (Parc National du Niokolo-Koba), Coll. Daget and Milon, 48.0–55.9 mm (syntypes of A. rheophilus).