Holoplanktonic organisms are thought to have enormous populations sizes, expansive geographic ranges, and low species diversity. Previous work suggests that pelagic gastropods are no exception to this generalization. However, most of these data are derived from temperate species, whereas the bulk of pteropod diversity occurs in the tropics. Here, we present DNA barcode data for COI focused on the under-studied limacinoid, gymnosome, and pseudothecosome pteropods collected from the coastal waters of Panama. We applied four molecular species delimitation approaches to determine the number of operational taxonomic units (OTUs) identifiable from our sequences and all data available from Barcode of Life Database (BOLD) and GenBank and compared these to the Barcode Index Number generated by the BOLD. Assemble Species by Automatic Partitioning (ASAP), Poisson Tree Processes (PTP), Bayesian Poisson Tree Processes (bPTP), and Generalized Mixed Yule Coalescent (GMYC) methods generally gave congruent results and suggest that pteropod diversity in the tropics is significantly underrepresented both in DNA sequence databases and in the number of named OTUs. Of 42 specimens collected and sequenced, we recovered 16 OTUs, only one of which belonged to an OTU already represented in sequence databases. Neighbor-joining trees including the previously published sequences show that in all three groups morphospecies can include genetically divergent lineages. Many very divergent (>15% from nearest neighbor) taxa are also still represented by only a single sequence, suggesting that there is a large amount of cryptic or pseudo-cryptic diversity still to be described. To aid this future endeavor, we include some preliminary 16S data derived from new pteropod-specific primers.

Abstract The democratisation of next‐generation sequencing has vastly increased the availability of sequencing data from metabarcoding. However, to effectively prepare these metabarcoding data for subsequent analysis, researchers must consistently apply several different bioinformatic tools—including those which denoise reads, cluster sequences and assign taxonomic identities. This often creates a bioinformatics bottleneck in workflows for non‐specialists due to obstacles around: (a) integrating different tools, (b) the inability to easily modify and rerun bioinformatic pipelines involving non‐scripted (‘point‐and‐click’) elements and (c) the multiple outputs that may be required of a single dataset (e.g. amplicon sequence variants [ASVs] and operational taxonomic units [OTUs]), which often results in users running pipelines multiple times. Here, we introduce SimpleMetaPipeline, an open‐source bioinformatics pipeline implemented in R, which addresses these obstacles. SimpleMetaPipeline integrates the most robust and commonly used existing bioinformatic tools in a single reproducible pipeline, with a streamlined choice of parameters, to generate a sequence data table containing alternative clustering and assignment options. SimpleMetaPipeline accepts demultiplexed paired‐end and single reads from multiple sequencing runs. We describe the pipeline and demonstrate how alternative annotations enable the easy implementation of multi‐algorithm agreement tests to strengthen inferences. SimpleMetaPipeline represents a valuable addition to the existing library of pipelines, providing easy and reproducible bioinformatics, including a range of commonly desired clustering and assignment options, such as OTUs and ASVs.

Previous research focusing on pelagic gastropods in the open ocean has demonstrated that many morphospecies comprise multiple distinct clades or cryptic species that can be distinguished with DNA sequence data. To date, the genetic diversity of the pelagic gastropod fauna of the tropical East Pacific, especially in shallow coastal waters, remains largely unexplored. To document the diversity of pterotracheoids (formerly heteropods) from the coastal waters of the Bay of Panama, we collected, photographed and sequenced fragments of mitochondrial cytochrome c oxidase subunit I (COI) and 16S ribosomal DNA for 60 atlantids, 3 carinariids and 6 pterotracheids. In addition to the COI barcode, our results include the first published 16S sequences for these groups. We found 11 operational taxonomic units (OTUs): 9 in the genus Atlanta, 1 Carinaria and 1 Firoloida. We report the presence of Atlanta oligogyra (Clades A and B), Atlanta turriculata, Atlanta lesueurii, Atlanta helicinoidea (Clade B), Atlanta plana, Atlanta echinogyra, Atlanta inflata and Atlanta frontieri through comparisons of our sequences with previously published sequences. We did not find Atlanta gaudichaudi, Atlanta inclinata, Atlanta tokiokai, Atlanta gibbosa, Atlanta peronii, or Oxygyrus inflatus, which have previously been reported from the region. Haplotype networks and estimates of phi(ST) illustrate how some species show population differentiation across the tropical Indo-Pacific region, whereas others show little apparent population structure. For example, the most common haplotypes of A. inflata and of A. turriculata occur in the Indian Ocean, the Central and West Pacific and the tropical East Pacific, whereas individuals of A. frontieri from the Indian Ocean do not share haplotypes with individuals of A. frontieri from the Pacific Ocean. Analyses were limited by sample sizes, but these data suggest that population genetics approaches may be useful for reconstructing population histories of these important, but overlooked, components of the plankton.

Accurate identification of fishes is essential for understanding their biology and to ensure food safety for consumers. DNA barcoding is an important tool because it can verify identifications of both whole and processed fishes that have had key morphological characters removed (e.g., filets, fish meal); however, DNA reference libraries are incomplete, and public repositories for sequence data contain incorrectly identified sequences. During a nine-year sampling program in the Philippines, a global biodiversity hotspot for marine fishes, we developed a verified reference library of cytochrome c oxidase subunit I (COI) sequences for 2,525 specimens representing 984 species. Specimens were primarily purchased from markets, with additional diversity collected using rotenone or fishing gear. Species identifications were verified based on taxonomic, phenotypic, and genotypic data, and sequences are associated with voucher specimens, live-color photographs, and genetic samples catalogued at Smithsonian Institution, National Museum of Natural History. The Biodiversity of Philippine Marine Fishes dataset is released herein to increase knowledge of species diversity and distributions and to facilitate accurate identification of market fishes.

Peppermint shrimp resembling Lysmata vittata, a species native to the Indo-West Pacific, were found in the lower Chesapeake Bay and adjacent coastal embayments in 2013, representing the first recorded introduction of this species in the northwestern Atlantic. Conflicting morphological descriptions, inconsistent morphological terminology, and limited molecular data (i.e., unresolved taxonomy), as well as the destruction of the type material of L. vittata, created uncertainty regarding proper identification. We provide the first phylogeny incorporating individuals from across the presumed native and introduced range of L. vittata. Morphological and phylogenetic analyses clearly indicate L. vittata represents a species complex of two widely divergent groups: 1) "Bruce Type" with a uniramous dorsal antennule that agrees with A.J. redescription of L. vittata, and 2) "Rauli Type" with a one-article accessory branch on the dorsal antennule that agrees most closely with the junior synonym L. rauli. Given the taxonomic ambiguity surrounding L. vittata, we designate the individual used by A.J. Bruce to redescribe L. vittata and incorporated in our analyses as a neotype to fix the identity of this species. We therefore identify introduced North American and New Zealand populations as L. vittata sensu stricto and postulate that the native range spans temperate/subtropical East Asia. These data suggest that L. rauli is a valid species, which includes a possible undescribed sister species. We confirm the presence of L. californica in New Zealand, the first non-native record for this species. We also provide data suggesting L. dispar may be more widespread in the Indo-West Pacific than currently known and consider L. lipkei to be a likely junior synonym.

Peppermint shrimp resembling Lysmata vittata Stimpson, 1860, a species native to the Indo-West Pacific, were found in the lower Chesapeake Bay and adjacent coastal embayments in 2013, representing the first recorded introduction of this species in the northwestern Atlantic. Conflicting morphological descriptions, inconsistent morphological terminology, and limited molecular data (i.e., unresolved taxonomy), as well as the destruction of the type material of L. vittata , created uncertainty regarding proper identification. We provide the first phylogeny incorporating individuals from across the presumed native and introduced range of L. vittata . Morphological and phylogenetic analyses clearly indicate L. vittata represents a species complex of two widely divergent groups: 1) “Bruce Type” with a uniramous dorsal antennule that agrees with A.J. Bruce’s 1990 redescription of L. vittata , and 2) “Rauli Type” with a one-article accessory branch on the dorsal antennule that agrees most closely with the junior synonym L. rauli Laubenheimer & Rhyne, 2010. Given the taxonomic ambiguity surrounding L. vittata , we designate the individual used by A.J. Bruce to redescribe L. vittata and incorporated in our analyses as a neotype to fix the identity of this species. We therefore identify introduced North American and New Zealand populations as L. vittata sensu stricto and postulate that the native range spans temperate/subtropical East Asia. These data suggest that L. rauli is a valid species, which includes a possible undescribed sister species. We confirm the presence of L. californica Stimpson, 1866 in New Zealand, the first non-native record for this species. We also provide data suggesting L. dispar Hayashi, 2007 may be more widespread in the Indo-West Pacific than currently known and consider L. lipkei Okuno & Fiedler, 2010 to be a likely junior synonym.

Biodiversity assessments are critical for setting conservation priorities, understanding ecosystem function and establishing a baseline to monitor change. Surveys of marine biodiversity that rely almost entirely on sampling adult organisms underestimate diversity because they tend to be limited to habitat types and individuals that can be easily surveyed. Many marine animals have planktonic larvae that can be sampled from the water column at shallow depths. This life stage often is overlooked in surveys but can be used to relatively rapidly document diversity, especially for the many species that are rare or live cryptically as adults. Using DNA barcode data from samples of nemertean worms collected in three biogeographical regions—Northeastern Pacific, the Caribbean Sea and Eastern Tropical Pacific—we found that most species were collected as either benthic adults or planktonic larvae but seldom in both stages. Randomization tests show that this deficit of operational taxonomic units collected as both adults and larvae is extremely unlikely if larvae and adults were drawn from the same pool of species. This effect persists even in well-studied faunas. These results suggest that sampling planktonic larvae offers access to a different subset of species and thus significantly increases estimates of biodiversity compared to sampling adults alone. Spanish abstract is available in the electronic supplementary material.

Peppermint shrimp resembling Lysmata vittataStimpson, 1860, a species native to the Indo-West Pacific, were found in the lower Chesapeake Bay and adjacent coastal embayments in 2013, representing the first recorded introduction of this species in the northwestern Atlantic. Conflicting morphological descriptions, inconsistent morphological terminology, and limited molecular data (i.e., unresolved taxonomy), as well as the destruction of the type material of L. vittata, created uncertainty regarding proper identification. We provide the first phylogeny incorporating individuals from across the presumed native and introduced range of L. vittata. Morphological and phylogenetic analyses clearly indicate L. vittata represents a species complex of two widely divergent groups: 1) “Bruce Type” with a uniramous dorsal antennule that agrees with A.J. Bruce’s 1990 redescription of L. vittata, and 2) “Rauli Type” with a one-article accessory branch on the dorsal antennule that agrees most closely with the junior synonym L. rauliLaubenheimer & Rhyne, 2010. Given the taxonomic ambiguity surrounding L. vittata, we designate the individual used by A.J. Bruce to redescribe L. vittata and incorporated in our analyses as a neotype to fix the identity of this species. We therefore identify introduced North American and New Zealand populations as L. vittata sensu stricto and postulate that the native range spans temperate/subtropical East Asia. These data suggest that L. rauli is a valid species, which includes a possible undescribed sister species. We confirm the presence of L. californicaStimpson, 1866 in New Zealand, the first non-native record for this species. We also provide data suggesting L. disparHayashi, 2007 may be more widespread in the Indo-West Pacific than currently known and consider L. lipkeiOkuno & Fiedler, 2010 to be a likely junior synonym.

The morphological diversity of marine annelid larvae is stunning. Although many of the larval forms have been categorized as trochophores or modified trochophores there are a few groups with distinctive larval features that make them easy to distinguish from other annelid larvae. We collected 252 annelid larvae from the plankton, with particular emphasis on oweniids, polygordiids, and thalassematids (i.e., echiurans) and sequenced fragments of their cytochrome c oxidase subunit 1 and 16S ribosomal RNA genes. We found 6 oweniid, 5 polygordiid, and 8 thalassematid OTUs. Thalassematids were found only in samples from the Pacific, while oweniids were found only in Caribbean samples. Among the oweniids we found two distinct morphotypes, one with a narrow, cloche shape and another that had a wider and more rectangular shape with clearly-developed lappets. Among the polygordiids we identified one larva as Polygordius eschaturus and several larvae as Polygordius jenniferae . All larvae, except for the Polygordius eschaturus, which was at a stage to early to make a determination, were endolarvae. Among the thalassematids we identified larvae of Ochetostoma edax, and found 7 unidentified OTUs. Finally, 150 miscellaneous polychaete larvae were sequenced, representing ~76 OTUs. 4 rostraria larvae from the Caribbean, whose sequences confirm the long-held assumption that they are amphinomids, could not be identified to species. In total only 5% of these OTUs could be identified to species with known sequences and most could not be identified to genus or even family with reasonable certainty. It is clear that metabarcoding approaches, although useful for documenting the number of OTUs, will be of limited use for identifying them until reference databases have improved their coverage for neotropical marine annelids.

Across plants and animals, genome size is often correlated with life-history traits: large genomes are correlated with larger seeds, slower development, larger body size and slower cell division. Among decapod crustaceans, caridean shrimps are among the most variable both in terms of genome size variation and life-history characteristics such as larval development mode and egg size, but the extent to which these traits are associated in a phylogenetic context is largely unknown. In this study, we examine correlations among egg size, larval development and genome size in two different genera of snapping shrimp, Alpheus and Synalpheus, using phylogenetically informed analyses. In both Alpheus and Synalpheus, egg size is strongly linked to larval development mode: species with abbreviated development had significantly larger eggs than species with extended larval development. We produced the first comprehensive dataset of genome size in Alpheus (n = 37 species) and demonstrated that genome size was strongly and positively correlated with egg size in both Alpheus and Synalpheus. Correlated trait evolution analyses showed that in Alpheus, changes in genome size were clearly dependent on egg size. In Synalpheus, evolutionary path analyses suggest that changes in development mode (from extended to abbreviated) drove increases in egg volume; larger eggs, in turn, resulted in larger genomes. These data suggest that variation in reproductive traits may underpin the high degree of variation in genome size seen in a wide variety of caridean shrimp groups more generally.

Surveys of marine invertebrate larvae are a powerful tool for the detection of hidden diversity (Barber & Boyce, 2006; Collin, et al., 2019; Collin et al., 2019a, 2019b; Mahon et al., 2010). They may be particularly effective at capturing deep-water, small, or infaunal species that are difficult to collect as adults by use of standard sampling techniques, and thus provide an independent assessment of diversity that complements species lists based solely on studies of adults (Collin, et al., 2020; Mahon et al., 2010; Sewell & Jury, 2011). In some cases, larval sampling can uncover unexpectedly high diversity (Collin, et al., 2019; Mahon et al., 2010). Here we apply this approach to compare the diversity of adult and larval asteroids from the coastal waters of Panama. A review of Caribbean biodiversity from the Census of Marine Life documented 116 species of adult asteroids in the wider Caribbean (Miloslavich et al., 2010: supplemental data table S8). Twenty-five species were reported for the Caribbean coast of Panama, less Received: 21 April 2020 | Accepted: 31 August 2020 DOI: 10.1111/ivb.12303

Surveys of larval diversity consistently increase biodiversity estimates when applied to poorly documented groups of marine invertebrates such as phoronids and hemichordates. However, it remains to be seen how helpful this approach is for detecting unsampled species in well-studied groups. Echinoids represent a large, robust, well-studied macrofauna, with low diversity and low incidence of cryptic species, making them an ideal test case for the efficacy of larval barcoding to discover diversity in such groups. We developed a reference dataset of DNA barcodes for the shallow-water adult echinoids from both coasts of Panama and compared them to DNA sequences obtained from larvae collected primarily on the Caribbean coast of Panama. We sequenced mitochondrialcytochrome c oxidase subunit I(COI) for 43 species of adult sea urchins to expand the number and coverage of sequences available in GenBank. Sequences were successfully obtained forCOIand16Sribosomal DNA from 272 larvae and assigned to 17 operational taxonomic units (OTUs): 4 from the Pacific coast of Panama, where larvae were not sampled as intensively, and 13 from the Caribbean coast. Of these 17 OTUs, 13 were identified from comparisons with our adult sequences and belonged to species well documented in these regions. Another larva was identified from comparisons with unpublished sequences in the Barcode of Life Database (BOLD) as belonging toPseudoboletia, a genus scarcely known in the Caribbean and previously unreported in Panama. Three OTUs remained unidentified. Based on larval morphology, at least two of these OTUs appeared to be spatangoids, which are difficult to collect and whose presence often goes undetected in standard surveys of benthic diversity. Despite its ability to capture unanticipated diversity, larval sampling failed to collect some species that are locally common along the Caribbean coast of Panama, such asLeodia sexiesperforata,Diadema antillarum, andClypeaster rosaceus.

The diversity of tropical marine invertebrates is poorly documented, especially those groups for which collecting adults is difficult. We collected the planktonic tornaria larvae of hemichordates (acorn worms) to assess their hidden diversity in the Neotropics. Larvae were retrieved in plankton tows from waters of the Pacific and Caribbean coasts of Panama, followed by DNA barcoding of mitochondrial cytochrome c oxidase subunit I (COI) and 16S ribosomal DNA to estimate their diversity in the region. With moderate sampling efforts, we discovered six operational taxonomic units (OTUs) in the Bay of Panama on the Pacific coast, in contrast to the single species previously recorded for the entire Tropical Eastern Pacific. We found eight OTUs in Bocas del Toro province on the Caribbean coast, compared to seven species documented from adults in the entire Caribbean. All OTUs differed from each other and from named acorn worm sequences in GenBank by >10% pairwise distance in COI and >2% in 16S. Two of our OTUs matched 16S hemichordate sequences in GenBank: one was an unidentified or unnamed Balanoglossus from the Caribbean of Panama, and the other was an unidentified ptychoderid larva from the Bahamas. The species accumulation curves suggest that nearly all the species have been collected and only one more species might still remain undetected in the Pacific. In contrast, the Caribbean species accumulation curve suggests that further sampling could yield more than 10 additional OTUs. Tornaria from the 14 OTUs exhibited typical planktotrophic morphologies, and, in some cases, may be distinguished by differences in pigmentation and by the number of telotrochal ciliary bands, but in general, few diagnostic differences were detected.

Lingulids and discinids are the only brachiopods that exhibit life histories that include a feeding planktonic stage usually referred to as a larva. We collected planktotrophic brachiopod larvae from the Pacific and Caribbean coasts of Panama and took a DNA barcoding approach with mitochondrial cytochrome c oxidase subunit I (COI), mitochondrial ribosomal 16S, and nuclear ribosomal 18S genes to identify those larvae and to estimate their diversity in the region. We observed specimens from both coasts with distinct morphologies typical of lingulid and discinid larvae. COI and 16S were sequenced successfully for the lingulid larvae but failed consistently for all discinid larvae. 18S was sequenced successfully for larvae from both families. Sequence data from each gene revealed one lingulid operational taxonomic unit (OTU) from Bocas del Toro on the Caribbean coast, and one lingulid OTU from the Bay of Panama on the Pacific coast. These OTUs differed by >20% for COI, >10% for 16S and 0.5% for 18S. Both OTUs clustered with GenBank sequences of Glottidia species, the only genus of lingulids in the Americas, but were distinct from G. pyramidata the only species reported for the Caribbean. Analysis of 18S sequence data for discinid larvae recovered 2 OTUs, one exclusively from the Pacific and one with a mixture of Pacific and Caribbean larvae. The 18S marker does not provide enough resolution to distinguish between species, and comparisons with GenBank sequences suggest that one OTU includes Pelagodiscus species, while the other may include Discradisca species. When compared with other marine invertebrates, our surveys of brachiopod larvae through DNA barcoding show relatively low levels of diversity for Panama.

Phoronid larvae, actinotrochs, are beautiful and complicated organisms which have attracted as much, if not more, attention than their adult forms. We collected actinotrochs from the waters of the Pacific and Caribbean coasts of Panama, and used DNA barcoding of mtCOI, as well as 16S and 18S sequences, to estimate the diversity of phoronids in the region. We discovered three operational taxonomic units (OTUs) in the Bay of Panama on the Pacific coast and four OTUs in Bocas del Toro on the Caribbean coast. Not only did all OTUs differ from each other by >10% pairwise distance in COI, but they also differed from all phoronid sequences in GenBank, including the four species for which adults have been reported for the Pacific of Panama, Phoronopsis harmeri, Phoronis psammophila, Phoronis muelleri, and Phoronis hippocrepia. In each ocean region, one common OTU was more abundant and occurred more frequently than other OTUs in our samples. The other five OTUs were relatively rare, with only one to three individuals collected during the entire project. Species accumulation curves were relatively flat but suggest that at least one more species is likely to be present at each site. Actinotrochs from the seven sequenced OTUs had morphologies typical of species with non-brooded planktotrophic development and, in some cases, may be distinguished by differences in pigmentation and the arrangement of blood masses. We found one larva with morphology typical of brooded planktotrophic larvae for which sequencing failed, bringing the total number of potential species detected to eight and representing >50% of the adult species currently recognized globally.

Advances in biodiversity genomic sequencing will increasingly depend on the availability of DNA samples—and their quantifiable metadata—preserved in large institutional biorepositories that are discoverable to the scientific community. Improvements in sequencing technology constantly provide longer reads, such that longer fragment length, higher molecular weight, and overall “genome-quality” DNA (gDNA) will be desirable. Ideally, biorepositories should publish numerical scale measurements of DNA quality useful to the user community. However, the most widely used technique to evaluate DNA quality, the classic agarose gel, has yet to be quantified. Here we propose a simple and economical method using open source image analysis software to make gDNA gel images quantifiable, and propose percentage of gDNA “greater thanXkb” as a standard of comparison, whereXis a band from any widely used DNA ladder with desirably large band sizes. We employ two metadata standards (“DNA Threshold” and “Percent above Threshold”) introduced as part of the Global Genome Biodiversity Network (GGBN) Darwin Core extension. We illustrate the method using the traditionally usedHindIII ladder and the 9,416 base-pair (bp) band as a standard. We also present data, for two taxa, a vertebrate (fish) and an invertebrate (crab), on how gDNA quality varies with seven tissue preservation methods, time since death, preservation method (i.e. buffers vs. cold temperatures), and storage temperature of various buffers over time. Our results suggest that putting tissue into a buffer prior to freezing may be better than directly into ultra-cold conditions.

We used 454 pyrosequencing to discover more than 3000 candidate microsatellite markers for three putative species of tadpole shrimp in the genus Triops. We selected 42 of these candidates, consisting of 12 systems identified in Triops longicaudatus “long,” 16 identified in Triops longicaudatus “short,” and 14 identified in Triops newberryi. Out of these potential loci we identified between five and eight polymorphic microsatellite systems per “species.” By implementing an additional cross amplification experiment with the microsatellite markers mentioned above and further testing 15 microsatellite markers initially identified in the European Triops cancriformis, we successfully transferred 17 microsatellite markers within the three North American Triops and found three new microsatellites, one polymorphic, that will amplify in T. cancriformis. The reported genetic methods are powerful molecular tools to acquire new detailed information at the interface of ecology, evolution and development, which will facilitate understanding phenotypic plasticity, breeding systems, and adaptation in this taxonomically-confusing group of “living fossils.”

The tropical shrimp genus Synalpheus includes the only eusocial marine animals. In much of the Caribbean, eusocial species have dominated the diverse fauna of sponge-dwelling shrimp in coral rubble for at least the past two decades. Here we document a recent, dramatic decline and apparent local extinction of eusocial shrimp species on the Belize Barrier Reef. Our collections from shallow reefs in central Belize in 2012 failed to locate three of the four eusocial species formerly abundant in the area, and showed steep declines in colony size and increases in frequency of queenless colonies prior to their disappearance. Concordant with these declines, several nonsocial, pair-forming Synalpheus species increased in frequency. The decline in eusocial shrimp is explained in part by disappearance of two sponge species on which they specialize. Eusocial shrimp collections from Jamaica in 2012 showed similar patterns of decline in colony size and increased queenlessness compared with prior Jamaican collections. The decline and local extinction of eusocial shrimp happened against a backdrop of changes in coral assemblages during recent decades, and may reflect changes in abundance and quality of dead coral substratum and succession of the diverse cryptic organisms living within it. These changes document potentially worrisome declines in a unique taxon of eusocial marine animals.

Tadpole shrimp are known to be important animals in the ecology of ephemeral wetlands. In the northern Chihuahuan Desert of North America, the tadpole shrimp fauna is composed of possibly three species in the genus Triops, which have variously been referred to as species, subspecies, and intraspecific variation. Our results support the presence of three morphologically distinct forms, which will be referred to herein as T. newberryi, T. longicaudatus "short," and T longicaudatus "long." We report analyses of Triops spp. sampled in summer 2008 from 14 natural playas and man-made flood retention ponds. Data were recorded on meristic counts and quantitative measurements of morphological features. We also sequenced portions of the mitochondrial COI and ND 1 genes for 72 shrimp, including individuals from all three morphological forms and for multiple ponds for each form where possible. The three forms were morphologically distinct for multiple characters and molecular analyses showed large differences in DNA nucleotide sequence and the presence of multiple unique amino acid substitutions in each form. Finally, prior literature suggests the three forms exhibit different reproductive systems, with populations of T. longicaudatus "long" thought to be gonochoric (equal sex ratios and obligate outcrossing), T. longicaudatus "short" having only self-fertilizing hermaphrodites, and T. newberryi being androdioecious, having both self-fertilizing hermaphrodites and males. While these three forms may be sufficiently distinct in morphology, mitochondria! DNA, and reproductive life history to warrant elevation to species level, additional geographical sampling and an examination of the various type specimens are necessary for a formal taxonomic revision.

Sampling of eight sites along the west coast of Barbados, West Indies, yielded 14 species of sponge-dwelling shrimps in the gambarelloides group of the genus Synalpheus, including one new species described here as Synalpheus microneptunus n. sp. The new species is a member of the S. paraneptunus Coutiere species complex and is distinguished from other species in that group by the combination of four carpal segments in the second pereopod, uropodal exopod with 2(nd) disto-lateral tooth smaller than the other two teeth and set in line with movable spine, and a small blade on the scaphocerite. Synalpheus microneptunus n. sp. is the smallest species in the complex (2.2-2.9 mm CL) and lives in small colonies, usually with fewer than 10 individuals, often with a single breeding female. Synalpheus thele Macdonald, Hultgren & Duffy is reported for the first time from outside its type locality in Jamaica. Sampling in Barbados produced fewer species than did similar efforts in Jamaica and Curacao, possibly due to the relatively isolated position of the island at the eastern (windward) edge of the Caribbean Sea.