Single-specimen systematics resolves the phylogeny and diversity conundrum of enigmatic crustacean y-larvae.

Resolving the evolutionary history of organisms is a major goal in biology. Yet for some taxa the diversity, phylogeny, and even adult stages remain unknown. The enigmatic crustacean "y-larvae" (Facetotecta) are one particularly striking example. Here, we use extensive video-imaging and single-specimen molecular sequencing of >200 y-larval specimens to comprehensively explore for the first time their evolutionary history and diversity. This integrative approach revealed five major clades of Facetotecta, four of which encompass a considerable larval diversity. Whereas morphological analyses recognized 35 y-naupliar "morphospecies", molecular species-delimitation analyses suggested the existence of between 88 and 127 species. The phenotypic and genetic diversity between the morphospecies suggests that a more elaborate classification than the current one-genus approach is needed. Morphology and molecular data were highly congruent at shallower phylogenetic levels, but no morphological synapomorphies could be unambiguously identified for major clades, which mostly comprise both planktotrophic and lecithotrophic y-nauplii. We argue that lecithotrophy arose several times independently whereas planktotrophic y-nauplii, which are structurally more similar across clades, most likely display the ancestral feeding mode of Facetotecta. We document a remarkably complex and highly diverse phylogenetic backbone for a taxon of larval marine crustaceans, the full life cycle of which remains a mystery.

Geographical and Temporal Structures of Legionella pneumophila Sequence Types in Comunitat Valenciana (Spain), 1998 to 2013.

Legionella pneumophila is an accidental human pathogen associated with aerosol formation in water-related sources. High recombination rates make Legionella populations genetically diverse, and nearly 2,000 different sequence types (STs) have been described to date for this environmental pathogen. The spatial distribution of STs is extremely heterogeneous, with some variants being present worldwide and others being detected at only a local scale. Similarly, some STs have been associated with disease outbreaks, such as ST578 or ST23. Spain is among the European countries with the highest incidences of reported legionellosis cases, and specifically, Comunitat Valenciana (CV) is the second most affected area in the country. In this work, we aimed at studying the overall diversity of Legionella pneumophila populations found in the period from 1998 to 2013 in 79 localities encompassing 23 regions within CV. To do so, we performed sequence-based typing (SBT) on 1,088 L. pneumophila strains detected in the area from both environmental and clinical sources. A comparison with the genetic structuring detected in a global data set that included 20 European and 7 non-European countries was performed. Our results reveal a level of diversity in CV that can be considered representative of the diversity found in other countries worldwide.

The emergence of lobsters: phylogenetic relationships, morphological evolution and divergence time comparisons of an ancient group (decapoda: achelata, astacidea, glypheidea, polychelida).

Lobsters are a ubiquitous and economically important group of decapod crustaceans that include the infraorders Polychelida, Glypheidea, Astacidea and Achelata. They include familiar forms such as the spiny, slipper, clawed lobsters and crayfish and unfamiliar forms such as the deep-sea and "living fossil" species. The high degree of morphological diversity among these infraorders has led to a dynamic classification and conflicting hypotheses of evolutionary relationships. In this study, we estimated phylogenetic relationships among the major groups of all lobster families and 94% of the genera using six genes (mitochondrial and nuclear) and 195 morphological characters across 173 species of lobsters for the most comprehensive sampling to date. Lobsters were recovered as a non-monophyletic assemblage in the combined (molecular + morphology) analysis. All families were monophyletic, with the exception of Cambaridae, and 7 of 79 genera were recovered as poly- or paraphyletic. A rich fossil history coupled with dense taxon coverage allowed us to estimate and compare divergence times and origins of major lineages using two drastically different approaches. Age priors were constructed and/or included based on fossil age information or fossil discovery, age, and extant species count data. Results from the two approaches were largely congruent across deep to shallow taxonomic divergences across major lineages. The origin of the first lobster-like decapod (Polychelida) was estimated in the Devonian (∼409-372 Ma) with all infraorders present in the Carboniferous (∼353-318 Ma). Fossil calibration subsampling studies examined the influence of sampling density (number of fossils) and placement (deep, middle, and shallow) on divergence time estimates. Results from our study suggest including at least 1 fossil per 10 operational taxonomic units (OTUs) in divergence dating analyses. [Dating; decapods; divergence; lobsters; molecular; morphology; phylogenetics.].

New mitogenomes of Runcinidae and Facelinidae: two understudied heterobranch families (Mollusca: Gastropoda).

Here, we present the mitochondrial sequences of two sea slugs (Heterobranchia): Runcina aurata and Facelina auriculata, the latter being the type species of the family. The mitochondrial genomes are 14,282 and 14,171bp in length, respectively, with a complete set of 13 PCGs, 2 rRNAs, and 22 tRNAs. None of the mitogenomes show gene reorganization, keeping the standard mitogenomic structure of Heterobranchia. Nucleotide composition differs significantly between them, with R. aurata showing the most AT-rich mitogenome (25.7% GC content) reported to date in Heterobranchia, and F. auriculata showing a rich GC content (35%) compared with other heterobranch mitochondrial genomes.

Novel molecular resources for single-specimen barcoding of enigmatic crustacean y-larvae.

Despite discovery more than 100years ago and documented global occurrence from shallow waters to the deep sea, the life cycle of the enigmatic crustacean y-larvae isincompletely understood and adult forms remain unknown. To date, only 2 of the 17 formally described species, all based on larval stages, have been investigated using an integrative taxonomic approach. This approach provided descriptions of the morphology of the naupliar and cyprid stages, and made use of exuvial voucher material and DNA barcodes. To improve our knowledge about the evolutionary history and ecological importance of y-larvae, we developed a novel protocol that maximises the amount of morpho-ecological and molecular data that can be harvested from single larval specimens. This includes single-specimen DNA barcoding and daily imaging of y-nauplii reared in culture dishes, mounting of the last naupliar exuviae on a slide as a reference voucher, live imaging of the y-cyprid instar that follows, and fixation, DNA extraction, amplification and sequencing of the y-cyprid specimen. Through development and testing of a suite of new primers for both nuclear and mitochondrial protein-coding and ribosomal genes, we showcase how new sequence data can be used to estimate the phylogeny of Facetotecta. We expect that our novel procedure will help to unravel the complex systematics of y-larvae and show how these fascinating larval forms have evolved. Moreover, we posit that our protocols should work on larval specimens from a diverse array of moulting marine invertebrate taxa.

The Catalan initiative for the Earth BioGenome Project: contributing local data to global biodiversity genomics.

The Catalan Initiative for the Earth BioGenome Project (CBP) is an EBP-affiliated project network aimed at sequencing the genome of the >40 000 eukaryotic species estimated to live in the Catalan-speaking territories (Catalan Linguistic Area, CLA). These territories represent a biodiversity hotspot. While covering less than 1% of Europe, they are home to about one fourth of all known European eukaryotic species. These include a high proportion of endemisms, many of which are threatened. This trend is likely to get worse as the effects of global change are expected to be particularly severe across the Mediterranean Basin, particularly in freshwater ecosystems and mountain areas. Following the EBP model, the CBP is a networked organization that has been able to engage many scientific and non-scientific partners. In the pilot phase, the genomes of 52 species are being sequenced. As a case study in biodiversity conservation, we highlight the genome of the Balearic shearwater Puffinus mauretanicus, sequenced under the CBP umbrella.

Integrative Taxonomy of Cyclocyprididae Kaufmann, 1900 (Ostracoda: Podocopa) with Description of a New Genus and Species.

The two widespread ostracod genera Cypria Zenker, 1854 and Physocypria Vávra, 1897 are traditionally distinguished based on the presence or absence of tubercles on the right valve margin. However, recent research based on soft body parts has uncovered new cryptic genera within Cypria and Physocypria. Following this line of research, a new Cyclocyprididae genus and species, Vizcainocypria viator gen. nov. sp. nov., is here described from individuals collected in rice fields and wetlands of the Iberian Peninsula. Vizcainocypria is compared with Cypria, Physocypria, Dentocypria Savatenalinton, 2017, Keysercypria Karanovic, 2011, Brasilocypria Almeida et al., 2023, and Claudecypria Almeida et al., 2023 based on morphological evidence. Besides the presence or absence of tubercles on the right valve, these genera can be distinguished according to their mandibular palp, second thoracopod, caudal ramus, and male hemipenis. Molecular analyses using mitochondrial (COX1), and nuclear (28S rDNA) genes provide further support for the differentiation of Cypria, Dentocypria, Physocypria and Vizcainocypria gen. nov. The present study highlights the importance of using an integrative taxonomy approach, combining shell and soft-body parts morphology and molecular data, to characterize the rich diversity of freshwater ostracods.

Identification of Phyllosoma Larvae of the Lobsters Acantharctus and Biarctus (Crustacea: Scyllaridae) from the SW Indian Ocean.

The accurate assignment of cryptic larvae to species-level is a key aspect of marine ecological research and can be achieved through integrated molecular and morphological studies. A combination of two mitochondrial markers (COI and 16S) and a detailed morphological analysis was used to identify phyllosoma larvae of slipper lobster (Scyllaridae) species collected during a survey in the SW Indian Ocean. Two morphotypes were tentatively assigned to Acantharctus ornatus and Biarctus pumilus, both genera for which the larval morphology was unknown. Morphological revision of an adult specimen used to generate the putative A. ornatus sequences in GenBank revealed that it was misidentified and corresponds to B. dubius. The final phyllosoma stage of B. pumilus and subfinal and final stages of A. ornatus were described, clarifying prior misidentifications in the literature. Scyllarid biodiversity in the SW Indian Ocean is underestimated and sampling of deeper water layers is recommended to complete current knowledge of species and larval stages present in the region.

Phylogeographic patterns of decapod crustaceans at the Atlantic-Mediterranean transition.

Comparative multispecies studies allow contrasting the effect of past and present oceanographic processes on phylogeographic patterns. In the present study, a fragment of the COI gene was analyzed in seven decapod crustacean species from five families and with different bathymetric distributions. A total of 769 individuals were sampled along the Atlantic-Mediterranean transition area in order to test the effect of three putative barriers to gene flow: Strait of Gibraltar, Almeria-Oran Front and Ibiza Channel. A significant effect of the Strait of Gibraltar was found in the crabs Liocarcinus depurator and Macropipus tuberculatus. The Ibiza Channel had a significant effect for L. depurator. However, the Almeria-Oran front was not found to have a significant effect on any of the studied species. Higher levels of population structure were found in shallow-water species, although the number of species sampled should be increased to obtain a conclusive pattern. The haplotypes within the different species coalesced at times that could be related with past climatic events occurring before, during and after the last glacial maximum. Given the large diversity of phylogeographic patterns obtained within decapods, it is concluded that both historical and contemporary processes (marine current patterns, bathymetry and life-history traits) shape the phylogeographic patterns of these crustaceans.

Mislabeling and nomenclatorial confusion of Typhlotanais sandersi Kudinova-Pasternak, 1985 (Crustacea: Tanaidacea) and establishment of a new genus.

Re-examination of historical collections allowed us to resolve the taxonomic status of Typhlotanais sandersi Kudinova-Pasternak, 1985, originally described based on a single specimen from Great-Meteor Seamount. The holotype of this species was considered lost and the species redescribed based on a second specimen from the type locality by Blazewicz-Paszkowycz (2007a), who placed Ty. sandersi on a newly established genus Typhlamia. Thorough morphological analysis of Typhlamia and Typhlotanais species and recently obtained genetic data of typhlotanaids from N Atlantic and NW Pacific waters allow us to conclude that the redescription of Ty. sandersi by Blazewicz-Paszkowycz (2007a) was based on a wrongly labelled specimen that, rather than a type of Ty. sandersi, represents in fact a new species of Typhlamia. The morphological comparison of the type species of Typhlotanais (Ty. aequiremis) with all 'long-bodied' typhlotanaid taxa with rounded pereonites margins (i.e., Typhlamia, Pulcherella, Torquella), and the use of genetic evidence, support the establishment of a new genus to accommodate: Ty. sandersi, Ty. angusticheles Kudinova-Pasternak, 1989, and a third species from N Atlantic waters, that is described here for the first time. Current knowledge on 'long-bodied' typhlotanaids with rounded pereonites is summarised and a taxonomical key for their identification provided.

MICROSATELIGHT--pipeline to expedite microsatellite analysis.

MICROSATELIGHT is a Perl/Tk pipeline with a graphical user interface that facilitates several tasks when scoring microsatellites. It implements new subroutines in R and PERL and takes advantage of features provided by previously developed freeware. MICROSATELIGHT takes raw genotype data and automates the peak identification through PeakScanner. The PeakSelect subroutine assigns peaks to different microsatellite markers according to their multiplex group, fluorochrome type, and size range. After peak selection, binning of alleles can be carried out 1) automatically through AlleloBin or 2) by manual bin definition through Binator. In both cases, several features for quality checking and further binning improvement are provided. The genotype table can then be converted into input files for several population genetics programs through CREATE. Finally, Hardy-Weinberg equilibrium tests and confidence intervals for null allele frequency can be obtained through GENEPOP. MICROSATELIGHT is the only freely available public-domain software that facilitates full multiplex microsatellite scoring, from electropherogram files to user-defined text files to be used with population genetics software. MICROSATELIGHT has been created for the Windows XP operating system and has been successfully tested under Windows 7. It is available at http://sourceforge.net/projects/microsatelight/.

Parasitoids (Hymenoptera) of Mealybug Pests (Hemiptera: Pseudococcidae) from Southern Brazil: Molecular and Morphological Characterization.

Parasitoids of three mealybug pests (Hemiptera: Pseudococcidae), Planococcus ficus (Signoret), Pseudococcus sociabilis Hambleton, and Pseudococcus viburni (Signoret) have been identified for the first time in Brazil. Mealybugs were collected in fruit-growing areas along southern Brazil during 2013-2016. An integrative approach, combining morphological and molecular methods, was used to identify the Brazilian parasitoids to the species level. Fifteen species were recorded, including 14 primary parasitoids belonging to Encyrtidae and Platygastridae and a single secondary parasitoid species belonging to Signiphoridae. The encyrtid parasitoids Acerophagus flavidulus (Brèthes), Anagyrus calyxtoi Noyes and Zaplatycerus sp., and the signiphorid secondary parasitoid Chartocerus axillaris De Santis are reported for the first time in Brazil.

A new species of Cypris (Crustacea: Ostracoda) from the Iberian Peninsula and the Balearic Islands, with comments on the first ostracod named using the Linnean system.

The genus Cypris, considered the oldest ostracod generic name erected using the Linnean system, comprises a reduced number of large-bodied species, mostly found in Africa and Asia. Only six of them are known to occur in Europe. Here we describe a new species, Cypris pretusi sp. nov., collected in small temporary streams and ponds along the Eastern Iberian Peninsula and Minorca (Balearic Islands). The new species is very close to the type species of the genus, Cypris pubera O.F. Müller, 1776, but differs from it in having a set of smaller subequal spines on the posterior edge of the valves, by the absence of conspicuous spines along the front edge, and by the beak-like frontal shape of its carapace in dorsal view, similar to Cypris decaryi Gauthier, 1933. Soft parts are very similar to the type species, but it differs in having shorter swimming setae on the second antennae. Molecular analyses of the COX1 region support its status as a species distinct from C. pubera and closer to Cypris bispinosa Lucas, 1849, also providing evidence for a separation of C. pubera s.l. in two clades, one of which is here considered to correspond to Cypris triaculeata Daday, 1892. We discuss the relationships of C. pretusi sp. nov. to other members of the genus and its possible origin from nearby biogeographic regions (probably Africa or Asia) and provide a key to species of Cypris found in Europe. We also discuss the relationship between Monoculus concha pedata (= M. conchaceus), the first ostracod named by Linnæus, and Cypris pubera, the type species of the genus, described by Müller in 1776 and considered by him the same species as the one first named by Linnæus.

Rapid radiation in spiny lobsters (Palinurus spp) as revealed by classic and ABC methods using mtDNA and microsatellite data.

BACKGROUND: Molecular tools may help to uncover closely related and still diverging species from a wide variety of taxa and provide insight into the mechanisms, pace and geography of marine speciation. There is a certain controversy on the phylogeography and speciation modes of species-groups with an Eastern Atlantic-Western Indian Ocean distribution, with previous studies suggesting that older events (Miocene) and/or more recent (Pleistocene) oceanographic processes could have influenced the phylogeny of marine taxa. The spiny lobster genus Palinurus allows for testing among speciation hypotheses, since it has a particular distribution with two groups of three species each in the Northeastern Atlantic (P. elephas, P. mauritanicus and P. charlestoni) and Southeastern Atlantic and Southwestern Indian Oceans (P. gilchristi, P. delagoae and P. barbarae). In the present study, we obtain a more complete understanding of the phylogenetic relationships among these species through a combined dataset with both nuclear and mitochondrial markers, by testing alternative hypotheses on both the mutation rate and tree topology under the recently developed approximate Bayesian computation (ABC) methods. RESULTS: Our analyses support a North-to-South speciation pattern in Palinurus with all the South-African species forming a monophyletic clade nested within the Northern Hemisphere species. Coalescent-based ABC methods allowed us to reject the previously proposed hypothesis of a Middle Miocene speciation event related with the closure of the Tethyan Seaway. Instead, divergence times obtained for Palinurus species using the combined mtDNA-microsatellite dataset and standard mutation rates for mtDNA agree with known glaciation-related processes occurring during the last 2 my. CONCLUSION: The Palinurus speciation pattern is a typical example of a series of rapid speciation events occurring within a group, with very short branches separating different species. Our results support the hypothesis that recent climate change-related oceanographic processes have influenced the phylogeny of marine taxa, with most Palinurus species originating during the last two million years. The present study highlights the value of new coalescent-based statistical methods such as ABC for testing different speciation hypotheses using molecular data.

Phylogenetic relationships between spiny, slipper and coral lobsters (Crustacea, Decapoda, Achelata).

Molecular data can aid in the resolution of conflicting hypotheses generated through difficulties in the interpretation of morphological data and/or an incomplete fossil record. Moreover, the reconstruction of phylogenetic relationships using molecular data may help to trace back the origin of morphological innovations which had a major impact on the radiation of a taxonomical group. In this work, different nuclear (18S, 28S, and H3) and mitochondrial (16S and COI) gene regions were sequenced in a total of 35 Achelatan species to test conflicting hypotheses of evolutionary relationships within the Achelata infraorder and solve the taxonomic disagreements in the group. The combined molecular dataset strongly supports the hypothesis that Achelata is a monophyletic group composed of two main families: Palinuridae and Scyllaridae. Synaxidae is found to be a polyphyletic group, which should be included within Palinuridae. Consequently, our results indicate that the origin of the stridulating organ occurred only once during Achelata evolution. Finally, the two main clades found within the Scyllaridae are in agreement with previous inferences based on adult morphological data. The dating of divergence of Achelata obtained with a relaxed-clock model is compatible with previous hypotheses of a Triassic origin of the Achelata.

Deep ocean seascape and Pseudotanaidae (Crustacea: Tanaidacea) diversity at the Clarion-Clipperton Fracture Zone.

Understanding the diversity and spatial distribution of benthic species is fundamental to properly assess the impact of deep sea mining. Tanaidacea provide an exceptional opportunity for assessing spatial patterns in the deep-sea, given their low mobility and limited dispersal potential. The diversity and distribution of pseudotanaid species is characterized here for the Clarion and Clipperton Fractures Zone (CCZ), which is the most extensive deposit field of metallic nodules. Samples were taken from the Belgian, German and French license areas, but also from the APEI 3 (Area of Particular Environmental Interest 3) of the Interoceanmetal consortium associates. The combination of morphological and genetic data uncovered one new pseudotanaid genus (Beksitanais n. gen.) and 14 new species of Pseudotanais (2 of them virtual taxa). Moreover, our results suggest that spatial structuring of pseudotanaid diversity is correlated with deep-sea features, particularly the presence of fractures and seamount chains crossing the CCZ. The presence of geographical barriers delimiting species distributions has important implications for the establishment of protected areas, and the APEI3 protected area contains only one third of the total pseudotanaid species in CCZ. The specimen collection studied here is extremely valuable and represents an important first step in characterizing the diversity and distribution of pseudotanaids within the Tropical Eastern Pacific.

Pancrustacean Evolution Illuminated by Taxon-Rich Genomic-Scale Data Sets with an Expanded Remipede Sampling.

The relationships of crustaceans and hexapods (Pancrustacea) have been much discussed and partially elucidated following the emergence of phylogenomic data sets. However, major uncertainties still remain regarding the position of iconic taxa such as Branchiopoda, Copepoda, Remipedia, and Cephalocarida, and the sister group relationship of hexapods. We assembled the most taxon-rich phylogenomic pancrustacean data set to date and analyzed it using a variety of methodological approaches. We prioritized low levels of missing data and found that some clades were consistently recovered independently of the analytical approach used. These include, for example, Oligostraca and Altocrustacea. Substantial support was also found for Allotriocarida, with Remipedia as the sister of Hexapoda (i.e., Labiocarida), and Branchiopoda as the sister of Labiocarida, a clade that we name Athalassocarida (="nonmarine shrimps"). Within Allotriocarida, Cephalocarida was found as the sister of Athalassocarida. Finally, moderate support was found for Hexanauplia (Copepoda as sister to Thecostraca) in alliance with Malacostraca. Mapping key crustacean tagmosis patterns and developmental characters across the revised phylogeny suggests that the ancestral pancrustacean was relatively short-bodied, with extreme body elongation and anamorphic development emerging later in pancrustacean evolution.

Integrative taxonomy methods reveal high mealybug (Hemiptera: Pseudococcidae) diversity in southern Brazilian fruit crops.

The Serra Gaúcha region is the most important temperate fruit-producing area in southern Brazil. Despite mealybugs (Hemiptera: Pseudococcidae) infesting several host plants in the region, there is a lack of information about the composition of species damaging different crops. A survey of mealybug species associated with commercial fruit crops (apple, persimmon, strawberry and grapes) was performed in Serra Gaúcha between 2013 and 2015, using both morphology and DNA analyses for species identification. The most abundant species were Pseudococcus viburni (Signoret), found on all four host plant species, and Dysmicoccus brevipes (Cockerell), infesting persimmon, vines and weeds. The highest diversity of mealybug species was found on persimmon trees, hosting 20 different taxa, of which Anisococcus granarae Pacheco da Silva & Kaydan, D. brevipes, Pseudococcus sociabilis Hambleton and Ps. viburni were the most abundant. A total of nine species were recorded in vineyards. Planococcus ficus (Signoret) and Pseudococcus longispinus (Targioni Tozzetti) were observed causing damage to grapes for the first time. A single species, Ps. viburni, was found associated with apples, while both Ps. viburni and Ferrisia meridionalis Williams were found on strawberry. Four of the mealybug species found represent new records for Brazil.

Tomato root microbiota and Phytophthora parasitica-associated disease.

BACKGROUND: Interactions between pathogenic oomycetes and microbiota residing on the surface of the host plant root are unknown, despite being critical to inoculum constitution. The nature of these interactions was explored for the polyphagous and telluric species Phytophthora parasitica. RESULTS: Composition of the rhizospheric microbiota of Solanum lycopersicum was characterized using deep re-sequencing of 16S rRNA gene to analyze tomato roots either free of or partly covered with P. parasitica biofilm. Colonization of the host root surface by the oomycete was associated with a shift in microbial community involving a Bacteroidetes/Proteobacteria transition and Flavobacteriaceae as the most abundant family. Identification of members of the P. parasitica-associated microbiota interfering with biology and oomycete infection was carried out by screening for bacteria able to (i) grow on a P. parasitica extract-based medium (ii), exhibit in vitro probiotic or antibiotic activity towards the oomycete (iii), have an impact on the oomycete infection cycle in a tripartite interaction S. lycopersicum-P. parasitica-bacteria. One Pseudomonas phylotype was found to exacerbate disease symptoms in tomato plants. The lack of significant gene expression response of P. parasitica effectors to Pseudomonas suggested that the increase in plant susceptibility was not associated with an increase in virulence. Our results reveal that Pseudomonas spp. establishes commensal interactions with the oomycete. Bacteria preferentially colonize the surface of the biofilm rather than the roots, so that they can infect plant cells without any apparent infection of P. parasitica. CONCLUSIONS: The presence of the pathogenic oomycete P. parasitica in the tomato rhizosphere leads to a shift in the rhizospheric microbiota composition. It contributes to the habitat extension of Pseudomonas species mediated through a physical association between the oomycete and the bacteria.

DNA barcoding the phyllosoma of Scyllarides squammosus (H. Milne Edwards, 1837) (Decapoda: Achelata: Scyllaridae).

Scyllarides has the largest number of species with commercial importance within the Scyllaridae family. As for other achelate lobsters, however, little is known of the unique long-lived planktonic phyllosoma stages of any of these tropical and temperate species. Recently, a large and diverse collection of Scyllaridae phyllosoma, compiled from cruises along the Coral Sea and spanning several years, has been analysed. Molecular evidence from DNA-barcoding and phylogenetic analyses is provided here on the identity of S. squammosus phyllosoma larvae, including stages that were previously undescribed or poorly known. As a consequence, the growth and morphological changes that occur during the mid- to late-stages of S. squammosus larval development is now well-documented. Furthermore, an additional collection of S. squammosus larvae, described by Alain Michel and thought to be no longer extant, were discovered in the crustacean collection of the Muséum national d'Histoire naturelle, Paris. This new molecular and morphological information is complemented by a review of the literature. As a result, descriptions of key larval characters by a number of authors were evaluated and appear to suggest the existence of distinct groups of larvae within Scyllarides. From a combination of adult and larval morphology, and molecular data, the results presented here revealed inconsistencies with regard to the affinities of species assigned to Scyllarides. This new evidence will contribute to future studies addressing the phylogenetic relationships within the genus.