Epibiotic ciliate communities on the crayfish Procambarus (Austrocambarus) sp. cultivated in a rustic aquaculture in southern Mexico.

Rustic aquaculture systems represent a sustainable production alternative in the world, providing appropriate conditions for the establishment of epibiotic ciliates on crayfish. However, the impact of epibiotic ciliates on the aquaculture production has been insufficiently studied hitherto. The objective of this work was to identify the epibiotic ciliates on the surface of the crayfish Procambarus (Austrocambarus) sp. and analyze whether there is any correlation between seasonality and environmental conditions in an artificial pond in Chiapas, Mexico. The crayfish were collected during dry and rainy seasons to investigate their ciliate epibionts. Some physical and chemical parameters of the sampled water were measured and correlation matrix analyses were applied to detect possible relationships between the presence of ciliates and environmental parameters. Thirteen species belonging to the subclasses Peritrichia and Suctoria were identified. Significant differences were found for some environmental parameters and species recorded in both seasons. The present data could help to better understand the relationship bewteen the epibiotic ciliate community on crayfish and some environmental parameters in aquaculture systems.

Localization of multiple opsins in ocular and non-ocular tissues of deep-sea shrimps and the first evidence of co-localization in a rhabdomeric R8 cell (Caridea: Oplophoroidea).

Bioluminescence is a prevalent phenomenon throughout the marine realm and is often the dominant source of light in mesophotic and aphotic depth horizons. Shrimp belonging to the superfamily Oplophoroidea are mesopelagic, perform diel vertical migration, and secrete a bright burst of bioluminescent mucous when threatened. Species in the family Oplophoridae also possess cuticular light-emitting photophores presumably for camouflage via counter-illumination. Many species within the superfamily express a single visual pigment in the retina, consistent with most other large-bodied mesopelagic crustaceans studied to date. Photophore-bearing species have an expanded visual opsin repertoire and dual-sensitivity visual systems, as evidenced by transcriptomes and electroretinograms. In this study, we used immunohistochemistry to describe opsin protein localization in the retinas of four species of Oplophoroidea and non-ocular tissues of Janicella spinicauda. Our results show that Acanthephyra purpurea (Acanthephyridae) retinas possess LWS-only photoreceptors, consistent with the singular peak sensitivity previously reported. Oplophoridae retinas contain two opsin clades (LWS and MWS) consistent with dual-sensitivity. Oplophorus gracilirostris and Systellaspis debilis have LWS in the proximal rhabdom (R1-7 cells) and MWS2 localized in the distal rhabdom (R8 cell). Surprisingly, Janicella spinicauda has LWS in the proximal rhabdom (R1-7) and co-localized MWS1 and MWS2 opsin paralogs in the distal rhabdom, providing the first evidence of co-localization of opsins in a crustacean rhabdomeric R8 cell. Furthermore, opsins were found in multiple non-ocular tissues of J. spinicauda, including nerve, tendon, and photophore. These combined data demonstrate evolutionary novelty and opsin duplication within Oplophoridae, with implications for visual ecology, evolution in mesophotic environments, and a mechanistic understanding of adaptive counter-illumination using photophore bioluminescence.

Evaluating conserved domains and motifs of decapod gonadotropin-releasing hormone G protein-coupled receptor superfamily.

G protein-coupled receptors (GPCRs) are an ancient family of signal transducers that are both abundant and consequential in metazoan endocrinology. The evolutionary history and function of the GPCRs of the decapod superfamilies of gonadotropin-releasing hormone (GnRH) are yet to be fully elucidated. As part of which, the use of traditional phylogenetics and the recycling of a diminutive set of mis-annotated databases has proven insufficient. To address this, we have collated and revised eight existing and three novel GPCR repertoires for GnRH of decapod species. We developed a novel bioinformatic workflow that included clustering analysis to capture likely GnRH receptor-like proteins, followed by phylogenetic analysis of the seven transmembrane-spanning domains. A high degree of conservation of the sequences and topology of the domains and motifs allowed the identification of species-specific variation (up to ~70%, especially in the extracellular loops) that is thought to be influential to ligand-binding and function. Given the key functional role of the DRY motif across GPCRs, the classification of receptors based on the variation of this motif can be universally applied to resolve cryptic GPCR families, as was achieved in this work. Our results contribute to the resolution of the evolutionary history of invertebrate GnRH receptors and inform the design of bioassays in their deorphanization and functional annotation.

Disparate response of decapods to low pH: A meta-analysis of life history, physiology and behavior traits across life stages and environments.

We employed a meta-analysis to determine if the presumed resilience of decapods to ocean acidification extends to all biological aspects, environments, and life stages. Most response categories appeared unaffected by acidification. However, certain fitness-related traits (growth, survival, and, to some extent, calcification) were impacted. Acid-base balance and stress response scaled positively with reductions in pH, which maintains homeostasis, possibly at the cost of other processes. Juveniles were the only stage impacted by acidification, which is believed to reduce recruitment. We observed few differences in responses to acidification among decapods inhabiting contrasting environments. Our meta-analysis shows decapods as a group slightly to moderately sensitive to low pH, with impacts on some biological aspects rather than on all specific life stages or habitats. Although extreme pH scenarios may not occur in the open ocean, coastal and estuarine areas might experience lower pH levels in the near to medium future, posing potential challenges for decapods.

Regulation of early spermatogenesis in the giant prawn Macrobrachium rosenbergii by a GCL homolog†.

The germ cell-less gene is crucial for gonad development in various organisms. Early interventions in its expression suggested a regulatory role at the mitotic stages of spermatogenesis, and its early knockout resulted in complete sterility in Drosophila. Genomic and transcriptomic data available for the catadromous giant prawn Macrobrachium rosenbergii enabled the identification of a germ cell-less homolog for this species, which we termed MroGCL (mRNA accession number OQ533056). An open reading frame containing 494 amino acids and a typical evolutionarily conserved BTB/POZ domain suggests possible protein-protein interaction functions in keeping with the Drosophila germ cell-less protein. Genomic mapping of MroGCL showed a full length of 120 896 bases. Analysis of the temporal expression of MroGCL showed constant expression in early prawn embryonic and larval stages, but a significant increase 10 days after metamorphosis when crucial sexual differentiation processes occur in prawns. In adult animals, high expression was detected in the gonads compared to the somatic tissues. RNAi-based knock-down experiments showed that both the silenced and control groups reached advanced spermatogenic stages, but that there was a significant decrease in the yield of spermatozoa in about half of the silenced animals. This finding supports our hypothesis that MroGCL is crucial for mitosis during early stage spermatogenesis. In conclusion, this study contributes to the understanding of crustacean gonad development and provides a stepping stone in the development of environmentally valuable sterile crustacean populations.

Evaluation of Black Soldier Fly Larvae Meal on Growth, Body Composition, Immune Responses, and Antioxidant Capacity of Redclaw Crayfish (Cherax quadricarinatus) Juveniles.

This study investigated the effects of substituting fish meal (FM) with black soldier fly (Hermetia illucens) meal (BSM) on the growth performance, body composition, immune response, and antioxidant enzyme activity of juvenile redclaw crayfish (Cherax quadricarinatus). Four isonitrogenous (41%) and isolipidic (11%) diets (i.e., FM substitutes) were formulated from BSM prepared using larvae that were fed soybean meal (BSM-S), fishery byproducts (BSM-F), or pitaya (BSM-P). The experimental diets were fed twice daily to triplicate groups of juvenile redclaw crayfish (0.56 ± 0.04 g). After the feed trial, the FM and BSM-F groups exhibited significantly lower feed conversion ratios and significantly higher weight gain; specific growth rates; and concentrations of saturated fatty acids, highly unsaturated fatty acids, eicosapentaenoic acid, and docosahexaenoic acid in the muscle. Among the tested groups, the BSM-F group exhibited significantly enhanced immune responses and increased antioxidant enzyme activity (i.e., superoxide dismutase, phenoloxidase, and glutathione peroxidase); the BSM-P group exhibited a significantly higher feed intake and hepatopancreatic index; and the FM group exhibited a significantly higher muscle body index and apparent digestibility for the dry matter of crude protein. The findings indicate that the juvenile redclaw crayfish fed BSM-F achieved the highest weight gain among the groups.

Antidepressants - The new endocrine disruptors? The case of crustaceans.

Antidepressants are high-volume pharmaceuticals that accumulate to concentrations in the µg·L-1 range in surface waters. The release of peptide hormones via neurosecretory cells appears as a natural target for antidepressants. Here I review research that suggests that antidepressants indeed disrupt endocrine signalling in crustaceans, by acting on the synthesis and release of neurohormones, such as crustacean hyperglycaemic hormone, moult inhibiting hormone and pigment dispersing hormone in decapods, as well as methyl farnesoate in Daphnids. Hence, antidepressants can affect hormonal regulation of physiological functions: increase in energy metabolism and activity, lowered ecdysteroid levels, potentially disrupting moult and somatic growth, reducing colour change capacity and compromising camouflage, as well as induction of male sex determination. Several studies further suggest effects of antidepressants on crustacean reproduction, but the hormonal regulation of these effects remains elusive. All things considered, a body of evidence strongly suggests that antidepressants are endocrine disrupting compounds in crustaceans.

Genome assemblies of two species of porcelain crab, Petrolisthes cinctipes and Petrolisthes manimaculis (Anomura: Porcellanidae).

Crabs are a large subtaxon of the Arthropoda, the most diverse and species-rich metazoan group. Several outstanding questions remain regarding crab diversification, including about the genomic capacitors of physiological and morphological adaptation, that cannot be answered with available genomic resources. Physiologically and ecologically diverse Anomuran porcelain crabs offer a valuable model for investigating these questions and hence genomic resources of these crabs would be particularly useful. Here, we present the first two genome assemblies of congeneric and sympatric Anomuran porcelain crabs, Petrolisthes cinctipes and Petrolisthes manimaculis from different microhabitats. Pacific Biosciences high-fidelity sequencing led to genome assemblies of 1.5 and 0.9 Gb, with N50s of 706.7 and 218.9 Kb, respectively. Their assembly length difference can largely be attributed to the different levels of interspersed repeats in their assemblies: The larger genome of P. cinctipes has more repeats (1.12 Gb) than the smaller genome of P. manimaculis (0.54 Gb). For obtaining high-quality annotations of 44,543 and 40,315 protein-coding genes in P. cinctipes and P. manimaculis, respectively, we used RNA-seq as part of a larger annotation pipeline. Contrarily to the large-scale differences in repeat content, divergence levels between the two species as estimated from orthologous protein-coding genes are moderate. These two high-quality genome assemblies allow future studies to examine the role of environmental regulation of gene expression in the two focal species to better understand physiological response to climate change, and provide the foundation for studies in fine-scale genome evolution and diversification of crabs.

Two new species of Parahesione (Annelida: Hesionidae) associated with ghost shrimps (Crustacea: Decapoda) and their phylogenetic relationships.

Two new species of Hesionidae, Parahesione pulvinata sp. nov. and Parahesione apiculata sp. nov. are described based on materials collected at tidal flats in Okinawa (Japan) from burrows of the ghost shrimps Neocallichirus jousseaumei and Glypturus armatus. The two new species are characterized by having eight enlarged cirri, dorsal cirrophores with dorsal foliose lobe and biramous parapodia, and by lacking median antenna. Parahesione apiculata sp. nov. has digitate lobes on the posterior margin of the dorsal foliose lobe (absent in P. pulvinata sp. nov.). The two new species were never found outside the ghost shrimp burrows, suggesting they are obligate symbionts. Phylogenetic analyses based on four concatenated genes suggest that the symbiotic lifestyle has evolved several times in Hesionidae.

Molecular evidence and differences in gonopod morphology lead to the recognition of a new species of the freshwater crab genus Candidiopotamon Bott, 1967 (Crustacea, Brachyura, Potamidae) from eastern Taiwan.

A new freshwater crab of the potamid genus Candidiopotamon Bott, 1967, is described from eastern Taiwan. Candidiopotamonpenglai sp. nov. is morphologically similar to C.rathbuni (De Man, 1914) from western Taiwan, but can be distinguished by the morphology of the male first gonopod (G1), as well as by their mitochondrial DNA (16S rRNA and COI genes). In the G1 of C.rathbuni, the subterminal segment shows a cline from robust in northern populations to slender in southern populations. In the G1 of C.penglaisp. nov., a distinctly larger and more distally directed keel-like projection is found on the distal inner edge of the terminal segment, with northern populations having an inward-curving subterminal segment and southern populations a straight subterminal segment. The genetic differentiation of the two species of Candidiopotamon within Taiwan is discussed, and morphological differences are compared. A key to the species of Candidiopotamon is also provided.

Molecular Phylogeny and Evolution of the Tuerkayana (Decapoda: Brachyura: Gecarcinidae) Genus Based on Whole Mitochondrial Genome Sequences.

Tuerkayana is of particular interest because it has been separated, in recent years, from Cardisoma and Discoplax but studies of its taxonomic status, especially from a whole mitochondrial genome perspective, have been lacking. In this study, the mitogenomes of four species (Tuerkayana magnum, Tuerkayana rotundum, Tuerkayana hirtipes, and Tuerkayana celeste) of Tuerkayana are sequenced and contrasted with other species in Brachyura for the first time. The phylogenetic tree of Brachyura, which includes 206 crab species (189 species of Brachyuran and 17 Anomura species) with a complete mitogenome, was constructed to evaluate the phylogenetic position of Tuerkayana and Gecarcinidae within Brachyuran, and explore the monophyly of Gecarcinidae. Furthermore, two single gene trees based on cox1 and 16SrRNA separately within interspecies of Gecarcinidae were reconstructed, providing molecular evidence for Tuerkayana and further clarifying the division of genera in Gecarcinidae. Based on the mitogenome dataset of 206 crabs, the branch-site model was utilized to explore selective pressure in individual codons with CodeML. The strong selective pressure shown in nad6 indicates that it may have played a significant role in the evolution of Gecarcinidae.

Notes on the Spreading of Penaeus aztecus Ives 1891 (Decapoda, Penaeidae) in the Mediterranean Sea and on Its Repeated Misidentifications in the Region.

The shrimp Penaeus aztecus, native to the western Atlantic, was first reported in the eastern Mediterranean Sea in 2010. New records, from different Mediterranean localities, multiplied in the following years. The accurate search of the literature on non-indigenous species discovered it was misidentified more than once as another alien shrimp, P. semisulcatus, native to the Indo-Pacific region, with the result that its earlier presence in the Black Sea went unnoticed. Morphological characteristics allowing the identification of these two species, the autochthonous P. kerathurus and two other alien Penaeus species present in the Mediterranean, are reprised. The present distribution of P. aztecus based on literature records and surveys carried out in the northern and central Adriatic between 2016 and 2021 is mapped. The unintentional transport of larvae carried in ballast water by transoceanic vessels departing from the U.S. East Coast is suggested as the most probable introduction pathway. The significance of the correct identification of non-indigenous species, a "Descriptor" adopted in the Marine Strategy Framework Directive for determining the good environmental status of marine waters in the European States, is emphasized.

Molecular Phylogeny of Thoracotreme Crabs Including Nine Newly Determined Mitochondrial Genomes.

Mitochondrial genomes are used widely for the molecular phylogenetic analysis of animals. Although phylogenetic analyses based on the mitogenomes of brachyurans often yield well-resolved phylogenies, most interfamilial phylogenetic relationships in Thoracotremata remain unclear. We determined nine new mitogenomes of Thoracotremata, including mitogenomes of Camptandriidae (Deiratonotus japonicus), Dotillidae (Ilyoplax integra, Ilyoplax pusilla, and Tmethypocoelis choreutes), Macrophthalmidae (Ilyograpsus nodulosus), Pinnotheridae (Arcotheres sp. and Indopinnixa haematosticta), Plagusiidae (Guinusia dentipes), and Percnidae (Percnon planissimum). Interestingly, Percnon planissimum (Percnidae) was found to possess ≥ 19 repeated sequences in the control region. The gene orders of Il. nodulosus, Arcotheres sp., and In. haematosticta were revealed to be unique among thoracotreme crabs. Although the results of Bayesian and maximum likelihood (ML) phylogenetic analyses of three datasets were incongruent, highly supported clades (PP ≥ 0.99 or BS ≥ 99%) were not contradictory among the analyses. All analyses suggested the paraphyly of Grapsoidea and Ocypodoidea, corroborating the findings of previous studies based on molecular phylogenies of thoracotreme crabs. The phylogenetic positions of symbiotic thoracotreme crabs, Pinnotheridae and Cryptochiridae, were highly supported (Pinnotheridae + Ocypodidae and Cryptochiridae + Grapsidae, respectively) for the Bayesian analyses but not for the ML analyses. Analyses of more thoracotreme species' mitogenome sequences in additional studies will further strengthen the framework for thoracotreme evolution.

Vibrio harveyi in a Caribbean spiny lobster (Panulirus argus) with hepatopancreas necrosis.

A Caribbean spiny lobster, Panulirus argus, was submitted for necropsy after a number of species-specific fatalities in a public aquarium. The hemolymph was opaque and did not clot and the hepatopancreas had multiple foci of necrosis centered on gram-negative bacteria. Pure cultures of Vibrio harveyi were isolated, identified initially by matrix laser desorption ionization-time of flight mass spectrometry, and confirmed by multilocus sequencing of the gyrB, recA, rpoA, and pyrH genes. As Caribbean spiny lobsters continue to be used for consumption and displayed in public aquariums, chronicling potential pathogens is warranted to inform differential diagnoses and to develop management strategies to reduce incidence of infectious disease in captive populations.

In situ microplastic ingestion by neritic zooplankton of the central Mexican Pacific.

Microplastics (MPs) are present across the global ocean and can be encountered by many species, including zooplankton. Although they fall within the size range of zooplankton prey, there are few studies on MPs ingestion carried out in situ. In this study, we analyzed MPs ingestion during two seasons (rainy and dry) of organisms from 5 taxonomic groups of zooplankton from two bays of the Mexican central Pacific: Manzanillo and Navidad. In total, 2643 individuals were analyzed, and of those 23 individuals contained MPs. The ingestion rate by taxonomic group was 1 MP/36 copepods (0.02), 1 MP/30 decapods-mysis (0.03), 1 MP/29 decapods-megalopa (0.03), and 1 MP/200 fish larvae (0.005). No plastics were found in chaetognaths, amphipods, or decapods-zoea. The average length of the ingested particles was 468.1 ± 113.8 µm, with a minimum of 15.6 and a maximum of 647.6 µm. All MPs >300 µm were fibers, with diameters <50 µm. Fragments were the most abundant MPs (54.2%), followed by fibers (34.2%) and spheres (11.4%). Statistical analyses showed no significant differences (p > 0.05) between the bays or seasons. Using RAMAN spectroscopy, it was possible to identify 6 different types of polymers, with poly (ethylene:propylene) being the most abundant (42.8%). This polymer is commonly used to manufacture plastic bags, ropes and fishing nets. The results confirm that certain zooplankton groups are consuming MPs and suggest that omnivorous species are more likely to ingest MPs, possibly due to their capacity for foraging flexibility and opportunistic feeding strategies. However, the ingestion of MPs cannot be attributed to a single factor; it is necessary to consider variables such as the sampling area, feeding strategy, size, and seasonality to understand the dynamics of MPs ingestion by zooplankton groups.

Microsporidia Ameson earli sp. n. and A. michaelis (Sprague 1965) infecting blue crabs Callinectes sapidus from shedding facilities in Louisiana.

During a survey for pathogens and commensals of blue crabs in commercial soft shell shedding facilities in Louisiana, we discovered an occurrence of microsporidiosis in two of forty examined crabs. Judging from spore shape and size, tissue tropism and external signs of muscle pathology, the causative agent of infections was identified as Ameson michaelis, a muscle-infecting species that has been repeatedly detected in populations of Callinectes sapidus in Louisiana since 1965. However, retrospective ultrastructural examination revealed that in one of Ameson-infected crabs, infection was caused by a parasite with ultrastructural characters not completely compliant with the ones of A. michaelis. The major difference was the absence of microtubule-like appendages attached to the exospore, typical of A. michaelis and other Ameson spp. SSUrDNA-inferred pairwise evolutionary distances between the novel species and other Ameson spp. ranged from 0.006 to 0.051; it was 0.039 in the case of A. michaelis. Hence, we describe here a new species in the genus Ameson, and name it after Prof. Earl Weidner, our colleague and friend, an outstanding microsporidiologist and the author of pioneer papers on the ultrastructure and physiology of A. michaelis.

New Insights into the Life-history Traits of the Night Shrimp Processa hemphilli (Caridea: Processidae) in the Western Atlantic.

We analyzed sexual dimorphism, sex ratio, relative growth, morphological sexual maturity, fecundity, embryo volume, and reproductive output of a population of the night shrimp Processa hemphilli from the Southeast coast of Brazil. Sampling was seasonally conducted, from June 2016 to April 2019 in Ubatuba, São Paulo. Our results showed the presence of sexual dimorphism in the body size, with females being larger than males. In addition, there was no difference in the overall sex ratio. However, it was possible to observe a skewed sex ratio to males in the smaller size classes, whereas females were more abundant in the larger size classes. The estimated carapace length at the onset of morphological sexual maturity of females and males was 3.6 mm and 3.2 mm, respectively. The mean fecundity of females was 285 ± 142 embryos (mean ± SD) embryos and was positively correlated with the carapace length. The embryo volume differed between the developmental stages with a constant increase in volume throughout the development between stages I and III. The mean reproductive output of females was 52.4%. Our results suggest that P. hemphilli have a pure search mating system, show sex allocation in the growth of body structures, and that females have high reproductive investment. All of these results represent the first insights into the life-history traits of the species in the western Atlantic and contribute to the knowledge about processid species.

Occurrence of Anthropogenic Debris in Three Commercial Shrimp Species from South-Western Ionian Sea.

Deep Sea environments represent the final collector of anthropogenic debris mainly represented by both plastic and non-plastic materials with different size. This led to potential contamination of deep marine fauna due to direct and indirect ingestion, representing a potential hazard for the species itself and for the final consumer. In this framework, the present study explored the occurrence of anthropogenic debris in the gastrointestinal tract of three Decapoda species of high commercial and ecological value (Parapenaeus longirostris, Aristeus antennatus, and Aristaeomorpha foliacea) from south-western Ionian Sea. After morphometrical measurements and sex determination, the gastrointestinal tract of 136 specimens were extracted and then chemically digested. A total of 230 low density microparticles were isolated, with a high frequency of occurrence in all the analyzed species (76% in P. longirostris, 70% in A. antennatus, and 83% in A. foliacea) mainly represented by fibers (92.6%) with a size between 0.10 and 0.49 mm, and with a dominance of the blue color. The results of the present study report for the first time the anthropogenic debris presence in the studied Decapoda from south-western Ionian Sea, highlighting the necessity to broaden the knowledge about anthropogenic debris pollution status in Mediterranean deep-sea species.

The pet trade as a source of non-native decapods: the case of crayfish and shrimps in a thermal waterbody in Hungary.

Ornamental aquaculture and the related pet industry are known to be important sources of non-native species worldwide. In the temperate zone, thermal waterbodies are attractive places for irresponsible owners to release unwanted freshwater pets including decapod crustaceans. Several non-native ornamental species have been reported in the thermal locality of Miskolctapolca (a suburb of Miskolc, Hungary). So we surveyed this site in March 2019-November 2021 to update local occurrence records and detect potentially newly released species. A well-established population of Neocaridina denticulata and the occurrence of Caridina cf. babaulti had previously been noted. However, for the first time at this site, we found the shrimps Atyopsis moluccensis, Caridina gracilirostris and C. multidentata, as well as the crayfish Procambarus virginalis, P. clarkii, Cherax quadricarinatus, C. boesemani and C. snowden, and several formally undescribed Cherax species originating from New Guinea. Furthermore, in most species, gravid females carrying eggs were also noticed. Three shrimps, A. moluccensis, C. gracilirostris and C. multidentata, were recorded for the first time in European wild. Further monitoring of this locality and better education of the general public regarding the risks associated with the release of non-native species are strongly recommended.

A comprehensive molecular phylogeny of the brachyuran crab superfamily Xanthoidea provides novel insights into its systematics and evolutionary history.

Maximum likelihood and Bayesian phylogenies for the brachyuran crab superfamily Xanthoidea were estimated based on three mitochondrial and four nuclear genes to infer phylogenetic relationships and inform taxonomy. Habitat data was then used in conjunction with several diversification rates analyses (BAMM, BiSSE, HiSSE, and FiSSE) to test evolutionary hypotheses regarding the diversification of xanthoid crabs. The phylogenies presented are the most comprehensive to date in terms of global diversity as they include all four constituent families (Xanthidae, Panopeidae, Pseudorhombilidae, and Linnaeoxanthidae) spanning all oceans in which xanthoid crabs occur. Six Xanthoidea families are recognised. Panopeidae and Xanthidae sensu stricto are the two largest family-level clades, which are reciprocally monophyletic. Pseudorhombilidae is nested within and is here treated as a subfamily of Panopeidae. Former subfamilies or tribes of Xanthidae sensu lato are basally positioned clades in Xanthoidea and are here assigned family-level ranks: Garthiellidae, Linnaeoxanthidae, Antrocarcinidae, and Nanocassiopidae. The genera Linnaeoxantho and Melybia were recovered in separate clades with Linnaeoxantho being sister to the family Antrocarcinidae, while Melybia was recovered within the family Panopeidae. The existing subfamily classification of Xanthidae and Panopeidae is drastically restructured with 20 xanthid and four panopeid subfamilies provisionally recognised. Diversification-time analyses inferred the origin of Xanthoidea and Garthiellidae in the Eocene, while the other families originated during the Oligocene. The majority of genus- and species-level diversification took place during the Miocene. Ancestral state reconstruction based on depth of occurrence (shallow vs. deep water) shows some ambiguity for the most recent common ancestor of Xanthoidea and Nanocassiopidae. The most recent common ancestors of Antrocarcinidae and Panopeidae were likely deep-water species, while those of Garthiellidae and Xanthidae were probably shallow-water species. Several shifts in net diversification rates were detected but they were not associated with depth-related habitat transitions.