The banded basslet, Lipogramma evides Robins & Colin, 1979, is shown to comprise two species: L. evides, which inhabits depths of 133–302 m, and a new species described here as Lipogramma levinsoni, which inhabits depths of 108–154 m and previously was considered to represent the juvenile of L. evides. A second new species of banded basslet, described here as Lipogramma haberi, inhabits depths of 152–233 m and was previously not reported in the literature. Morphologically, the three species differ in color patterns and modal numbers of gill rakers, whereas various other morphological features distinguish L. levinsoni from L. evides and L. haberi. DNA barcode data and multilocus, coalescent-based, species-delimitation analysis support the recognition of the three species. Phylogenetic analysis of mitochondrial and nuclear genetic data supports a sister-group relationship between the two deepest-living of the three species, L. evides and L. haberi, and suggests that the shallower L. levinsoni is more closely related to L. anabantoides Böhlke, 1960, which inhabits depths < 120 m. Evolutionary relationships within Lipogramma thus appear to be correlated with species depth ranges, an eco-evolutionary pattern that has been observed in other Caribbean marine teleosts and that warrants further investigation. The new species represent the eleventh and twelfth new fish species described in recent years from exploratory submersible diving in the Caribbean in the globally poorly studied depth zone of 50–300 m. This study suggests that there are at least two additional cryptic species of Lipogramma, which are being analyzed in ongoing investigations of Caribbean deep-reef ecosystems.

Manned submersible, cryptic species, integrative taxonomy, phylogeny, ocean exploration, Smithsonian Deep Reef Observation Project (DROP)

The western Atlantic family Grammatidae comprises small, usually brightly colored fishes in two genera, Gramma with four species and Lipogramma with eight (Robertson and Van Tassell 2015). Among other characters, the two genera are distinguished by the absence of a lateral line and presence of thickened, spinous, outer procurrent rays in Lipogramma (Mooi & Gill, 2002). The Banded Basslet, Lipogramma evides Robins & Colin, 1979, was described based on six specimens collected from Barbuda, Jamaica, Mexico, and Nicaragua. The original description also included observations of the species from Belize by Colin (1974). Subsequently, six additional specimens from the Bahamas were reported by Gilmore and Jones (1988). Robins and Colin (1979) noted differences in pigment pattern between adults and what they thought was a juvenile L. evides (Fig. 1A, B), in particular the presence of broader and more intense dark bands on the juvenile that completely encircle the body. Gilmore and Jones (1988) further commented on the presumed color differentiation between ontogenetic stages and noted that heavily banded “juveniles” (Fig. 1C) inhabit shallower waters (< 200 m) than adults (as deep as 250 m).

Figure 1. 

Previously published images of A Lipogramma evides, 34.4 mm SL, ANSP 134329, holotype, from Robins and Colin (1979: fig. 1) B Lipogramma levinsoni sp. n., 12.6 mm SL, ANSP 134332, as juvenile paratype of L. evides in Robins and Colin (1979: fig. 2) C Lipogramma levinsoni sp. n., 14.1 mm SL, IRCZM 107: 07660, as juvenile of L. evides in Gilmore and Jones (1988: fig. 3). Images reproduced with permission from Bulletin of Marine Science.

Exploratory submersible diving to 300 m in the southern and eastern Caribbean over the past several years by the Smithsonian Institution’s Deep Reef Observation Project (DROP) has resulted in the collection of over 50 specimens of “banded basslets” assignable to Lipogramma based on the absence of a lateral line and presence of spinous procurrent caudal-fin rays. That material includes individuals with both pigment patterns observed by previous authors and a third pigment pattern not previously described. Genetic and morphological analyses of individuals with those three pigment patterns suggested three distinct species and show that the heavily banded pattern is not an ontogenetic feature but diagnostic of a separate species. That species reaches a smaller maximum size than L. evides and has a shallower depth range. The other new species is similar in size and depth of occurrence to L. evides. Here we describe those two new species of Lipogramma, morphologically and genetically compare them with L. evides, and discuss depth distributions and evolutionary relationships of species of the genus.

Collecting and morphology. Basslets were collected using Substation Curaçao’s manned submersible Curasub (http://www.substation-curacao.com). The sub has two flexible, hydraulic arms, one of which is equipped with a quinaldine/ethanol-ejection system and the other with a suction hose. Anesthetized fish specimens were captured with the suction hose, which empties into a vented plexiglass cylinder attached to the outside of the sub. At the surface, the specimens were photographed, tissue sampled, and fixed in 10% formalin. Measurements were made weeks to months after fixation and subsequent preservation in 75% ethanol and were taken to the nearest 0.1 mm with dial calipers or an ocular micrometer fitted into a Wild stereomicroscope. Selected preserved specimens were later photographed to document preserved pigment pattern and X-rayed with a digital radiography system. Images of supraorbital pores and tooth-like structures on gill rakers were made using a Zeiss Axiocam on a Zeiss Discovery V12 SteREO microscope. Counts and measurements follow Hubbs and Lagler (1947). Specimens were cleared and stained following the protocol of Dingerkus and Uhler (1977). Symbolism for configuration of supraneural bones, anterior neural spines, and anterior dorsal pterygiophores follows Ahlstrom et al. (1976). USNM = Smithsonian Institution, National Museum of Natural History; ANSP = Academy of Natural Sciences, Philadelphia; IRCZM = Indian River Coastal Zone Museum, Harbor Branch Foundation, Fort Pierce, Florida; UF = University of Florida, Gainesville.

Molecular analyses. Tissue samples for 97 specimens assignable to eight species of Lipogramma were used for molecular analyses (Appendix 1). Tissues of L. rosea Gilbert, 1979 (in Robins and Colin 1979), L. regia Robins & Colin, 1979, and L. flavescens Gilmore & Jones, 1988 were not available. Tissues were stored in saturated salt-DMSO (dimethyl sulfoxide) buffer (Seutin et al. 1991). DNA extraction and cytochrome c oxidase subunit I (COI) DNA barcoding were performed for 96 specimens (i.e., for all available specimens except one L. anabantoides – Appendix 1) as outlined by Weigt et al. (2012). Four nuclear markers were amplified and sequenced—TMO-4C4, Rag1, Rhodopsin, and Histone H3—for 18 specimens of Lipogramma, and one or more of those genes was sequenced for an additional three specimens (Appendix 1). Primers and PCR conditions for the nuclear markers followed Lin and Hastings (2011, 2013). Sequences were assembled and aligned using Geneious v. 9 (Biomatters, Ltd., Aukland). A neighbor-joining (NJ) network was generated for the COI data using the K2P substitution model (Kimura 1980) in the tree-builder application in Geneious. Mean within- and between-species K2P genetic distances were calculated from the COI data in MEGA v. 7 (Kumar et al. 2015). Genetic distances were considered as corroborating morphology-based species delineation if the distances between species were ten or more times the intraspecific differences (Hebert et al. 2004). The alignments of COI and nuclear genes were concatenated and phylogeny was inferred using Bayesian Inference (BI) and Maximum Likelihood (ML), partitioning by gene. For the Bayesian analysis, substitution models and partitioning scheme were chosen using PartitionFinder (Lanfear et al. 2012) according to Bayesian Information Criterion scores. The chosen scheme had the following partitions and models: COI, HKY+I+G; Histone H3 plus Rhodospin, HKY+G; TMO-4C4, K80+G; Rag1, K80+G. All partitions in the ML analysis received a GTR-GAMMA substitution model. The BI phylogeny was inferred in the program MrBayes v. 3.2 (Ronquist et al. 2012) using two Metropolis-coupled Markov Chain Monte Carlo (MCMC) runs, each with four chains. The analysis ran for 10 million generations sampling trees and parameters every 1000 generations. Burn-in, convergence and mixing were assessed using Tracer (Rambaut and Drummond 2007) and by visually inspecting consensus trees from both runs. The ML analysis was done in the program RAxML v.8.2.9 (Stamatakis, 2014), using 20 initial random searches, and topological support was assessed using 1000 bootstrap replicates. Outgroups for the phylogenetic analysis included two species of Gramma and several other genera from the Ovalentaria sensuWainwright et al. (2012): Acanthemblemaria (Labrisomidae), Helcogramma (Tripterygiidae), Blenniella (Blenniidae), and Tomicodon (Gobiesocidae).

To corroborate the morphologically diagnosed species using our molecular data, we conducted a coalescent-based, Bayesian species-delimitation analysis (Yang and Rannala 2010, 2014). We used the computer program BP&P ver. 3.2 (Bayesian Phylogenetics and Phylogeography – Yang and Rannala [2010], Yang [2015]), which analyzes multi-locus DNA sequence alignments under the multispecies coalescent model (Rannala and Yang 2003). We used the five DNA alignments for the 21 Lipogramma specimens in BP&P, with each sequence in the alignments being assigned to one of eight groups a priori, based on the diagnostic morphological and coloration characters discussed in the ‘Morphological Comparisons’ section below. BP&P was then used to jointly infer a species tree and calculate posterior probabilities of different species-delimitation models containing either eight species, fewer than eight species (i.e. lumping multiple ‘morphological species’), or more than eight species (i.e. splitting ‘morphological species’ into multiple cryptic species).

Depth distributions. To evaluate depth distributions we searched FishNet2 (www.fishnet2.net) for all Lipogramma specimens that were identified to species and that included data on the depth of capture. For some specimens, capture depth was given as a range of possible depths, and in instances where this range was 50 m or narrower, we took the mean depth as a proxy for a point estimate of the exact depth of capture. Broader depth ranges of capture were excluded. Depth records for L. evides were only included for specimens whose identifications we confirmed to avoid possible confusion with one of the two new species described here. When combined with depth data from specimens from DROP collections, this search resulted in depth records for 278 identified specimens of Lipogramma. We also included depth records from 83 visual observations from DROP submersible dives, excluding those observations where there was uncertainty regarding identification of the three morphologically similar species (L. evides and the two species described here).

Accession numbers. GenSeq nomenclature (Chakrabarty et al. 2013) and GenBank accession numbers for DNA sequences derived in this study are presented along with museum catalog numbers for voucher specimens in Appendix 1.

Comments on Lipogramma evides. The type series of L. evides includes the holotype and five paratypes (Robins and Colin 1979). We examined specimens or photographs of specimens of the type series from ANSP and FMNH and conclude that all except one, ANSP 134332, 12.6 mm SL, represent L. evides. We also examined 31 specimens of Lipogramma evides that we recently collected at Curaçao and that range in size from 12.7–45.4 mm SL. Frequency distributions of pectoral-fin rays and gill rakers on the first arch are given in Table 2, an illustration of the holotype that was included in the original description of the species is shown in Fig. 1A, color patterns of live and recently deceased individuals are shown in Fig. 8, a photographed of a preserved paratype (ANSP 134330) is provided in Fig. 5C, secondary spines on gill rakers of the first arch are shown in Fig. 3, supraorbital pore pattern is shown in Fig. 4, and a photograph of a preserved juvenile is featured in Fig. 6.

Figure 8. 

Lipogramma evides A Aquarium photograph by Barry Brown, Substation Curacao BUSNM 276560, 45.3 mm SL, illustration by Grant Gilmore in Gilmore and Jones (1988: fig. 1) C and DUSNM 414885, 24.4 mm SL, photos by D. R. Robertson and C. C. Baldwin against black (C) and white (D) backgrounds.

The illustration of the holotype (Fig. 1A) shows a triangular-shaped bar on the posterior portion of the trunk that more closely resembles the shape of that bar in L. haberi (Fig. 7) than L. evides (Fig. 8). The pigment is too faded in the preserved holotype now to determine the shape of that bar, but we note that in specimens or photographs of the four paratypes of the type series that are actually specimens of L. evides, the posterior trunk bar is narrow (e.g., Fig. 5C). This suggests that the shape of the posterior trunk bar illustrated in the holotype of L. evides is either incorrectly drawn or represents an anomaly for the species. Lipogramma evides and L. haberi are easily distinguished by numbers of gill rakers on the first arch—15 or 16 in L. haberi, 19-22 (modally 20) in L. evides (Table 2). Our examination of the holotype confirms the count by Robins and Colin (1979) of 20 gill rakers in the holotype of L. evides. Furthermore, the triangular-shaped posterior trunk bar in L. haberi is very pale relative to the anterior trunk and head bars. Robins and Colin’s (1979) illustration of the holotype shows three body bars of equal intensity.

Colin’s (1974) observation of “Lipogramma sp.” at Glover’s Reef, Belize, was cited as L. evides by Robins and Colin (1979), and based on the recorded depths of observations, 165–180 m, we tentatively agree with this identification, as Belize is between Arrowsmith Bank and Nicaragua, where L. evides does occur. Lipogramma evides inhabits depths of 133-302 m, whereas L. levinsoni occurs from 103 to 154 m. However, Colin’s (1974) observed fish could have been L. haberi, which occurs from 152–233 m. Polanco et al. (2012) recorded several specimens of L. evides from the Coralinos Archipelago off Colombia, and based on the stated counts of 15–16 pectoral-fin rays and 20–21 gill rakers, those specimens are correctly identified. In addition to Colombia and the tentative Belize location, L. evides is known definitively from the type locality of Arrowsmith Bank in the Yucatan Peninsula, Nicaragua, southeast of Barbuda, and Curaçao (including Klein Curaçao). It was also observed but not collected in October 2016 by DRR and LT from the mini-submarine “Idabel” at 232–250 m depth adjacent to Half Moon Bay, Roatan, Honduras. It was not collected or observed on DROP submersible dives at Bonaire or Dominica. A list of L. evides material examined in this study is given in Appendix 2.

Morphological comparisons. Lipogramma levinsoni, L. evides, and L. haberi can be readily distinguished from all congeners in having three dark bars (one on the head, two on the trunk) on a white background vs. a brown body with a reddish head in L. anabantoides; a yellow body with one black bar on the head in L. flavescens; a purple head and yellow trunk in L. klayi Randall, 1963; a brown body with one broad white bar and multiple narrow orange bars in L. regia; a brownish body with about 12 thin dark bars in L. robinsi Gilmore, 1997; a pink head and trunk with a yellow stripe along the dorsal profile of the head in L. rosea; and a yellow head and trunk with three long iridescent blue stripes on the head in L. trilineata Randall, 1963. The major differences among L. levinsoni, L. evides, and L. haberi are summarized in Table 5. Lipogramma evides and L. haberi are morphologically similar and reach a similar maximum size (45.5 and 40.1 in our material, respectively). They are easy to distinguish from one another on the basis of number of gill rakers on the first arch (usually 20-21 in L. evides, 15-16 in L. haberi – Table 2) and by live and preserved color pattern (Figs 5, 7, 8). In life, L. haberi has a considerable amount of yellow as ground color and associated with the dark bars, whereas the ground color of L. evides is mostly white. In fresh and preserved specimens, the posterior trunk bar in L. haberi is broad and much wider at the top than the bottom, whereas in L. evides it is narrower and of uniform width. There is also a difference in the shape of the caudal fin of the two species, with L. haberi having a truncate fin with rounded lobe tips and L. evides having a slightly emarginate fin with pointed lobe tips.

Lipogramma levinsoni reaches a smaller maximum size than L. haberi and L. evides (largest specimen examined 28.3 mm SL) and differs in modal numbers of gill rakers on first arch and pectoral-fin rays (Table 2), supraorbital pore pattern (Fig. 4), and numerous aspects of color pattern (Figs 2, 5, 7, 8). In life, L. levinsoni is easily distinguished from L. haberi and L. evides by having an orange submarginal stripe on the dorsal fin (vs. yellow) and an orange submarginal stripe on the anal fin (vs. no stripe in L. haberi, a yellow submarginal stripe in L. evides). In preservative, L. levinsoni is easily distinguished from L. haberi and L. evides by the shape, size, and configuration of the dark body bars (Table 5).

Species delimitation and phylogeny. The neighbor-joining network (Suppl. material 1) shows eight distinct genetic lineages with an average within-lineage genetic distance of 0.002 substitutions/site and an average between-lineage genetic distance of 0.20 substitutions/site (Table 3). Considering between-lineage distances that are 10 or more times greater than within-lineage distances as indicative of distinct species (Hebert et al. 2014), genetic distances corroborate the recognition of the L. levinsoni and L. haberi lineages as species. Average between-lineage divergence for L. levinsoni is 19% (18% between L. levinsoni and the other two banded basslets, L. haberi and L. evides), whereas average within-lineage divergence is 0%. For L. haberi, average between-lineage divergence is 18% (11% between L. haberi and L. levinsoni/L. evides), whereas average within-lineage divergence is 0.2%.

The ML and BI analyses resulted in identical topologies, with most relationships supported by 1.0 posterior probability and 100% bootstrap support (Fig. 9). The BP&P analysis inferred a coalescent-based species-tree that was identical in topology to the ML and BI trees. In addition, the BP&P analysis provided overwhelming support for the presence of eight species of Lipogramma in our phylogeny (posterior probability 0.99981), including three distinct species of banded basslets (L. evides, L. haberi and L. levinsoni), indicating perfect congruence between molecular and morphology-based species delimitation criteria. Lipogramma trilineata and L. klayi, which have two of the shallowest depth ranges among Lipogramma species (Fig. 10), were recovered as sister species. This pair is sister to a larger clade comprising L. anabantoides + L. levinsoni (as sister species) and L. evides + L. haberi + two putative new species superficially resembling L. robinsi. Not surprising given their morphological similarity, L. evides, and L. haberi resolve as sister species. There is strong support for a clade comprising the four deepest-known species in our phylogeny—L. evides, L. haberi, and the two “L. robinsi” species (Figs 9, 10). Baldwin and Robertson (2014) found a similar evolutionary grouping of deep-water species in the serranid genus Liopropoma, and Tornabene et al. (2016a) found repeated invasions of deep-reef depths in the family Gobiidae with subsequent species radiations entirely within the deep-reef zone. Lipogramma flavescens, which also inhabits deep water (200–300 m, Fig. 10), may be part of this clade. A dark ocellus on the base of the soft dorsal fin appears to be a synapomorphy of the clade comprising L. anabantoides, L. levinsoni, L. evides, L. haberi, and the two “L. robinsi” species. Presence of this ocellus in L. flavescens and L. regia suggests that they may also belong to this group, but genetic samples of both are needed for phylogenetic analysis. Lipogramma flavescens may be closely related to L. haberi, as they share a narrow dark bar through the eye, yellow coloration, and low gill-raker count (15–16), and they inhabit similar deep-reef depths (152–233 m for L. haberi, 200–300 m for L. flavescens). If the evolutionary relationships of Lipogramma species are correlated with depth as our data suggest, and if Lipogramma regia, which is known only from depths < 100 m is a member of the clade diagnosed by a dark ocellus on the soft dorsal fin, it may be most closely related to L. anabantoides and L. levinsoni, which are known only from depths < 120 m (L. anabantoides) and < 154 m (L. levinsoni). Those three are the only known Lipogramma species with a modal pectoral-fin count of 17 (Gilmore and Jones 1988: Table 2, this study). We note that the addition to our molecular phylogeny of the four known species of Lipogramma that are currently unavailable for analysis (L. flavescens, L. regia, L. rosea, and L. robinsi) could change our hypotheses of relationships within the genus.

Figure 9. 

Bayesian Inference molecular phylogeny of Lipogramma based on combined mitochondrial and nuclear genes. Numbers of individuals analyzed for each species are given in Appendix 1, along with the genes sequenced for each individual. Topology is identical to that from Maximum Likelihood analysis. Support values are Bayesian posterior probabilities (above) and bootstrap values (below). Nodes without labels have 1.0 posterior probability and 100 bootstrap values. Photos or illustrations by C. C. Baldwin, D. R. Robertson, R. G. Gilmore, and C. R. Robins.

Figure 10. 

Depth distributions for species of Lipogramma. Photos or illustrations by C. C. Baldwin, D. R. Robertson, R. G. Gilmore, and Mooi and Gill (2002: fig. 9).

The two “L. robinsi” included here (Table 3, Figs 9, 10) are genetically distinct and superficially different from one another and from L. robinsi Gilmore, 1997. A more thorough investigation of those three taxa is in progress, after which a key to all Lipogramma species will be constructed.

Lipogramma is currently classified along with Gramma in the family Grammatidae based on a single synapomorphy in the arrangement of cheek musculature (Gill and Mooi 1993). Molecular data have failed to corroborate the monophyly of the Grammatidae (Betancur-R et al. 2013, Mirande 2016); rather, those data suggest that Lipogramma and Gramma are each related to different taxa within the diverse Ovalentaria. Relationships within the Ovalentaria have proven difficult to resolve with traditional molecular markers (Betancur-R et al. 2013), molecular markers plus some morphological characters (Mirande 2016), and phylogenomic data (Eytan et al. 2015). Potential close relatives of Lipogramma based on molecular data include blennioids, cichlids, plesiopids, pseudochromids, and Pholidichthys. Some of these groups have been previously linked to either Lipogramma, Gramma, or both, based on shared morphological characters, but the homologies of many of these characters have been questioned (Gill and Mooi 1993). At present, the phylogenetic position of Lipogramma is unresolved.

Ecology and life history. Little is known about community structure on deep reefs, including food networks. Although an analysis of the diet of banded basslets based on stomach contents is beyond the scope of this study, the gastrointestinal tract of the cleared and stained L. evides (USNM 434771) contained numerous individuals of a planktonic foraminiferan that was tentatively identified by Smithsonian Curator of Planktic Foraminifera Brian Huber as Globorotalia manardii (d’Orbigny) – Fig. 11A. Two items found in the gastrointestinal tract of L. levinsoni (USNM 406140) appear to be a diatom (possibly Coscinodiscus eccentricus Ehrenberg, Huber pers. comm, Fig. 11B) and a parasitic nematode (identification by Smithsonian Curator of Invertebrate Zoology Jon Norenberg and Assistant Professor of Biology at Virginia Military Institute Ashleigh Smythe, Fig. 11C). Future investigations of diets of deep-reef fish species are planned.

Figure 11. 

Items from stomachs of deep-reef Lipogramma: A Planktonic foraminiferan, possibly Globorotalia manardii, from L. evides, USNM 434771, collected at 174 m B Diatom, possibly Coscinodiscus eccentricus, from L. levinsoni sp. n., USNM 406140, collected between 137 and 146 m C Parasitic Nematoda from same specimen as B. Photos by A. Nonaka and L. Tornabene.

The broad Caribbean distributions of L. levinsoni and L. evides (Table 5) suggest a pelagic larval stage capable of dispersal, so it is perplexing that there are no records of Lipogramma or Gramma larvae from plankton tows (Asoh and Yoshikawa 1996, Hardy 2006). Thresher (1980) noted that L. trilineata constructs nests within algae in aquaria settings, and Asoh and Yoshikawa (1996) described similar nesting behavior in Gramma loreto Poey, 1868. The apparent restricted geographic distribution of L. haberi (Table 5) could indicate that some species have limited dispersal capabilities; however, the paucity of faunal investigations of deep-reef ecosystems may mask a larger geographic distribution for that species. Interestingly, Leis et al. (2012) calculated swimming speed for reared G. loreto larvae and found that the actual and relative critical speed (Ucrit) were so low that for most of the pelagic larval duration their ability to influence their dispersal by horizontal swimming would be much less than that of many other tropical fish species. Further study of the early life history of Lipogramma is needed, including exploring the possibility that planktonic dispersal in the genus may be limited.

Adults and juveniles of the banded basslet, L. evides, were previously recognized as different ontogenetic color patterns of a single species. This study shows that the juvenile color pattern belongs to a cryptic species, described here as L. levinsoni. This study also documents the first known juveniles of L. evides, which share the color pattern of adults. A second new species that is morphologically similar to L. evides, L. haberi, is also described. These three basslet species are confined to deep-reef depths, but they stratify such that L. levinsoni occurs at shallower depths than L. evides and L. haberi. A molecular analysis of evolutionary relationships among available Lipogramma species reveals correlations between depth of occurrence and phylogeny, an eco-evolutionary pattern observed in other deep-reef Caribbean fishes that warrants further investigation. The two new basslets represent the eleventh and twelfth new fish species described in recent years from exploratory submersible diving by the Smithsonian’s Deep Reef Observation Project (DROP) to Caribbean depths of 300 m (Baldwin and Robertson 2013, 2014, 2015; Baldwin and Johnson 2014; Baldwin et al. 2016; Tornabene et al. 2016b, 2016c). Numerous other new fish and invertebrate species discovered by DROP await description, including the two putative new species identified in this study as morphologically similar to but distinct from L. robinsi. Considerably more effort is needed to adequately explore tropical deep reefs, diverse but largely overlooked global ecosystems.

We thank Laura Albini, Bruce Brandt, Barry Brown, Mary Brown, Cristina Castillo, Loretta Cooper, Tico Christiaan, Tommy Devine, Grant Gilmore, Brian Horne, Brian Huber, Dave Johnson, Rob Loendersloot, Caleb McMahon, Dan Mulcahy, Jon Norenberg, Diane Pitassy, Sandra Raredon, Rob Robins, Laureen Schenk, Adriane “Dutch” Schrier, Ian Silver-Gorges, Ashleigh Smythe and Barbara van Bebber for assistance in various ways with this study. Funding for the Smithsonian Institution’s Deep Reef Observation Project was provided internally by the Consortium for Understanding and Sustaining a Biodiverse Planet to CCB, the Competitive Grants for the Promotion of Science program to CCB and DRR, the Herbert R. and Evelyn Axelrod Endowment Fund for systematic ichthyology to CCB, and STRI funds to DRR. Externally the research was funded by National Geographic Society’s Committee for Research and Exploration to CCB (Grant #9102-12). This is Ocean Heritage Foundation/Curacao Sea Aquarium/Substation Curacao (OHF/SCA/SC) contribution number 27.

Specimens of Lipogramma evides examined in this study.

ANSP 134329, holotype, 34.4 mm SL, R/V Pillsbury Sta. 581, Mexico, Arrowsmith Bank, 21°05'N, 86°23'W, 146–265 m depth, 22 May 1967; ANSP 134330, n=2, paratypes, 28.0–32.0 mm SL, R/V Pillsbury Sta. 581, Mexico, Arrowsmith Bank, 21 05'N, 86 23'W, 146–265 m depth, 22 May 1967; ANSP 134331, paratype, 17.2 mm SL, R/V Pillsbury Sta. 969, southeast of Barbuda, 17°27.8'N, 61°41.1'W, 68–216 m depth, 20 July 1969; FMNH 82583, paratype, 34.5 mm SL, Nicaragua, 12°32'N, 82°25 ‘ W, 155 m depth, 23 May 1692; USNM 426801 25.1 mm SL, Curasub submersible, sta. CURASUB13-18, Curaçao, Playa Forti, Westpoint, 12.3679 N, 69.1553 W, no depth data, 15 August 2013, C. Baldwin, B. Brandt, A. Schrier, K. Johnson & C. DeForest; USNM 431410, 12.7 mm SL , Curasub submersible, sta. CURASUB14-07, Curaçao, in between Porto Marie and Daaibooi beaches, 12.202842 N, 69.089507 W, 123 m depth, 21 March 2014, C. Baldwin et al.; USNM 426746, 45.4 mm SL, tissue no. CUR13279, Curasub submersible, sta. CURASUB13-19, Curaçao, Playa Forti, Westpoint, 12.3679 N, 69.1553 W, 179 m depth, 15 August 2013, B. Van Bebber, N. Knowlton, A. Schrier & R. Sant; USNM 431408, 35.5 mm SL, Curasub submersible, sta. CURASUB14-02, Curaçao, off Substation Curaçao downline., 12.083197 N, 68.899058 W, no depth data available, 17 March 2014, B. Brooks et al.; USNM 410992, 43.0 mm SL, Curasub submersible, sta. CURASUB13-33, Caracas Baii and back to Substation Curaçao downline, 12.068 N, 68.873367 W, 215 m depth, 5 November 2013, C. Baldwin, B. Brandt, A. Schrier & C. Castillo; UF 238591, 34.5 mm SL, Curasub submersible, sta. CURASUB15-13, Northwest corner of Klein Curaçao, 11.998453 N, 68.651308 W, 182 m depth, 28 August 2015, C. Baldwin & B. Van Bebber; USNM 434771, 33.3 mm SL, cleared and stained, tissue no. CUR15091, Curasub submersible, sta. CURASUB15-12, northwest corner of Klein Curaçao, 11.998453 N, 68.651308 W, 174 m depth, 27 August 2015, B. Brandt & A. Schrier; USNM 434783, 17.9 mm SL, tissue no. CUR15103, Curasub submersible, sta. CURASUB15-13, Northwest corner of Klein Curaçao, 11.998453 N, 68.651308 W, 171 m depth, 28 August 2015, C. Baldwin & B. Van Bebber; UF 238590, 27.7 mm SL, tissue no. CUR15104, Curasub submersible, sta. CURASUB15-13, Northwest corner of Klein Curaçao, 11.998453 N, 68.651308 W, 172 m depth, 28 August 2015, C. Baldwin & B. Van Bebber; USNM 431313, 39.2 mm SL, tissue no. T1K003, Curasub submersible, sta. CURASUB14-03, Curaçao, west of Substation Curaçao downline, 12.083197 N, 68.899058 W, 177 m depth, 18 March 2014, C. Baldwin et al.; USNM 414886, 24.9 mm SL, tissue no. CUR12013, Curasub submersible, sta. CURASUB12-01, Curaçao, off of Substation Curaçao downline, 12.083197 N, 68.899058 W, 171 m depth, 21 May 2012, C. Baldwin, A. Schrier & B. Brandt; USNM 414889, 31.3 mm SL, tissue no. CUR12031, Curasub submersible, Curaçao, off of Substation Curaçao downline, 12.083197 N, 68.899058 W, no depth data available, 21 May 2012, C. Baldwin et al.; USNM 414883, 40.0 mm SL, tissue CUR12044, Curasub submersible, Curaçao, off of Substation Curaçao downline, 12.083197 N, 68.899058 W, no depth data available, 21 May 2012, C. Baldwin et al.; USNM 414884, 32.0 mm SL, tissue no. CUR12050, Curasub submersible, sta. CURASUB12-11, Curaçao, off of Substation Curaçao downline, 12.083197 N, 68.899058 W, 164 m depth, 6 August 2012, B. Brandt, C. Baldwin, A. Schrier & A. Driskell; USNM 414887, 31.3 mm SL, tissue CUR12078, Curasub submersible, Curaçao, off of Substation Curaçao downline, 12.083197 N, 68.899058 W, no depth data available, 21 May 2012, C. Baldwin et al.; USNM 414890, 40.3 mm SL, tissue no. CUR12084, Curasub submersible, Curaçao, off of Substation Curaçao downline, 12.083197 N, 68.899058 W, no depth data available, 21 May 2012, C. Baldwin et al.; USNM 414888, 39.8 mm SL, tissue no. CUR12116, Curasub submersible, sta. CURASUB12-14, Curaçao, east of downline off Substation dock, 12.083197 N, 68.899058 W, 133 m depth, 9 August 2012, A. Schrier, B. Brandt, C. Castillo, A. Driskell & D. Robertson; USNM 414880, 29.0 mm SL, tissue no. CUR12117, Curasub submersible, sta. CURASUB12-14, Curaçao, east of downline off Substation dock, 12.083197 N, 68.899058 W, 134 m depth, 9 August 2012, A. Schrier, B. Brandt, C. Castillo, A. Driskell & D. Robertson; USNM 414882, 20.1 mm SL, tissue CUR12118, Curasub submersible, sta. CURASUB12-14, Curaçao, east of downline off Substation dock, 12.083197 N, 68.899058 W, 134 m depth, 9 August 2012, A. Schrier, B. Brandt, C. Castillo, A. Driskell & D. Robertson; USNM 414878, 24.1 mm SL, tissue no. CUR12276, Curasub submersible, sta. CURASUB12-16, Curaçao, west to Stella Maris and down, 154 m depth, 13 August 2012, A. Schrier, C. Baldwin & B. Van Bebber; USNM 414881, 21.1 mm SL, tissue no. CUR12280, 21.2 mm SL, Curasub submersible, sta. CURASUB12-17, Curaçao, East of downline off Substation Curaçao dock, 12.083197 N, 68.899058 W, 161 m depth, 13 August 2012, A. Schrier, B. Brandt, C. Castillo & D. Robertson; USNM 414885, 24.4 mm SL, tissue no. CUR12281, Curasub submersible, sta. CURASUB12-17, Curaçao, East of downline off Substation Curaçao dock, 12.083197 N, 68.899058 W, 161 m depth, 13 August 2012, A. Schrier, B. Brandt, C. Castillo & D. Robertson; USNM 414879, 25.0 mm SL, tissue no. CUR12288, Curasub submersible, sta. CURASUB12-16, Curaçao, west to Stella Maris and down, 154 m depth, 13 August 2012, A. Schrier, C. Baldwin & B. Van Bebber; USNM 414876, 41.7 mm SL, tissue no. CUR12353, Curasub submersible, no station data, off Substation Curaçao dock, 12.083197 N, 68.899058 W, no depth data available, 2012, Substation Curaçao crew; USNM 421602, 43.6 mm SL, tissue CUR13100, Curasub submersible, Curaçao, off Substation Curaçao, 12.083197 N, 68.899058 W, no depth data available; USNM 426769, 19.0 mm SL, tissue CUR13233, Curasub submersible, sta. CURASUB13-12, Curaçao, off downline at Substation Curaçao, 12.083197 N, 68.899058 W, 137–173 m depth, 7 August 2013, C. Baldwin, D. Robertson, C. Castillo & B. Van Bebber; USNM 426770, 13.7 mm SL, tissue no. CUR13234, Curasub submersible, sta. CURASUB13-12, Curaçao, off downline at Substation Curaçao, 12.083197 N, 68.899058 W, 137–173 m depth, 7 August 2013, C. Baldwin, D. Robertson, C. Castillo & B. Van Bebber; USNM 426771, 24.9 mm SL, tissue CUR13265, Curasub submersible, sta. CURASUB13-18, Curaçao, Playa Forti, westpoint, 12.3679 N, 69.1553 W, 137–164 m depth, 15 August 2013, C. Baldwin, B. Brandt, A. Schrier, K. Johnson & C. DeForest; USNM 426737, 19.3 mm SL, tissue CUR13266, Curasub submersible, sta. CURASUB13-18, Curaçao, Playa Forti, westpoint, 12.3679 N, 69.1553 W, 137–173 m depth, 15 August 2013, C. Baldwin, B. Brandt, A. Schrier, K. Johnson & C. DeForest; USNM 426709, 40.2 mm SL, tissue no. CUR13286, Curasub submersible, sta. CURASUB13-21, Curaçao, off Substation Curaçao downline, 12.083197 N, 68.899058 W, 171–201 m depth, 17 August 2013, C. Baldwin, A. Schrier, B. Brandt & A. Driskell; USNM 426722, 32.5 mm SL, tissue no. CUR13294, Curasub submersible, sta. CURASUB13-21, Curaçao, off Substation Curaçao downline, 12.083197 N, 68.899058 W, 171–201 m depth, 17 August 2013, C. Baldwin, A. Schrier, B. Brandt & A. Driskell.