Desenvolvimento tecnológico no cultivo de filosomas de lagostas palinurídeos / Technological development in the culture of phyllosoma of palinurid spiny lobster / Desarrollo tecnológico en lo cultivo de filosomas de langostas palinurídeos

O desenvolvimento larval completo dos estágios da filosoma da lagosta palinurídeo foi feito em condições de laboratório por vários pesquisadores. A duração dos estágios larvais parece ser específica em relação às espécies. Para as larvas filosomas recentes podem ser fornecidas náuplios de Artemia em combinação com pedaços de gônadas de mexilhões (Mytilus edulis). Gônadas de mexilhões podem ser utilizados exclusivamente após o segundo ou terceiro instar. Grande mortalidade pode ser observada no primeiro instar. A par disso, poucas espécies foram cultivadas devido ao seu dilatado e complexo desenvolvimento larval. A proposta desta revisão é resumir as informações sobre os estudos do cultivo de filosomas.(AU)

The complete larval development of the phylosoma stages of palinurid spiny lobster has been made under laboratory condition by several researchers. Duration of the larval stage seems to be species specific. For the early phyllosomas, food can be consisted of Artemia nauplii in combination with pieces of mussel (Mytilus edulis) gonad. Mussel gonad can be used exclusively after the 2nd or 3rd instar. Heavy mortalities can be noted at the 1st instar. Few spiny lobsters have been raised from egg to puerulus in the lab. Spiny lobsters have a long and complex larval development. The purpose of this review is to sumarize the studies on phyllosoma culture.(AU)

El desarrollo larvario completo de los estadios de filosoma de la langosta espinosa palinúrida ha sido realizado en condiciones de laboratorio por varios investigadores. A duración de las etapas larvais parece ser específica en relacion a las especies. Para las larvas filosomas recientes pueden ser ofrecidas nauplios de Artemia en cobinacion con pedazos de gónadas de mejillones (Mytilus edulis). Gónadas de mejillones pueden ser utilizados exclusivamente após o segundo el tercero instar. Grande mortalidad puede ser observada en lo primeiro instar. Al mismo tiempo, pocas especies han sido cultivadas debido a su dilatado y complejo desarrollo larval. La propuesta desta revisión es resumir la información sobre los estúdios sobre el cultivo de filosomas.(AU)

Mismatch of thermal optima between performance measures, life stages and species of spiny lobster.

In an ocean warming hotspot off south-east Australia, many species have expanded their ranges polewards, including the eastern rock lobster, Sagmariasus verreauxi. This species is likely extending its range via larval advection into Tasmanian coastal waters, which are occupied by the more commercially important southern rock lobster, Jasus edwardsii. Here, thermal tolerances of these lobster species at two life stages were investigated to assess how they may respond to warming ocean temperatures. We found that the pattern, optimum and magnitude of thermal responses differed between performance measures, life stages and species. Sagmariasus verreauxi had a warmer optimal temperature for aerobic scope and escape speed than J. edwardsii. However, J. edwardsii had a higher magnitude of escape speed, indicating higher capacity for escape performance. There were also differences between life stages within species, with the larval stage having higher variation in optimal temperatures between measures than juveniles. This inconsistency in performance optima and magnitude indicates that single performance measures at single life stages are unlikely to accurately predict whole animal performance in terms of life-time survival and fitness. However, combined results of this study suggest that with continued ocean warming, S. verreauxi is likely to continue to extend its distribution polewards and increase in abundance in Tasmania.

Multi-Tissue Transcriptome Analysis Identifies Key Sexual Development-Related Genes of the Ornate Spiny Lobster (Panulirus ornatus).

Sexual development involves the successive and overlapping processes of sex determination, sexual differentiation, and ultimately sexual maturation, enabling animals to reproduce. This provides a mechanism for enriched genetic variation which enables populations to withstand ever-changing environments, selecting for adapted individuals and driving speciation. The molecular mechanisms of sexual development display a bewildering diversity, even in closely related taxa. Many sex determination mechanisms across animals include the key family of "doublesex- and male abnormal3-related transcription factors" (Dmrts). In a few exceptional species, a single Dmrt residing on a sex chromosome acts as the master sex regulator. In this study, we provide compelling evidence for this model of sex determination in the ornate spiny lobster Panulius ornatus, concurrent with recent reports in the eastern spiny lobster Sagmariasus verreauxi. Using a multi-tissue transcriptomic database established for P. ornatus, we screened for the key factors associated with sexual development (by homology search and using previous knowledge of these factors from related species), providing an in-depth understanding of sexual development in decapods. Further research has the potential to close significant gaps in our understanding of reproductive development in this ecologically and commercially significant order.

Transcriptional profiling of spiny lobster metamorphosis reveals three new additions to the nuclear receptor superfamily.

BACKGROUND: The Crustacea are an evolutionarily diverse taxon which underpins marine food webs and contributes significantly to the global economy. However, our knowledge of crustacean endocrinology and development is far behind that of terrestrial arthropods. Here we present a unique insight into the molecular pathways coordinating crustacean metamorphosis, by reconciling nuclear receptor (NR) gene activity from a 12-stage, 3-replicate transcriptome in the ornate spiny lobster (Panulirus ornatus) during larval development. RESULTS: We annotated 18 distinct nuclear receptor genes, including three novel NRs which are upregulated prior to metamorphosis and have hence been named the "molt-associated receptors" (MARs). We also demonstrate the ecdysone-responsive expression of several known molt-related NRs including ecdysone receptor, fushi-tarazu-F1 and E75. Phylogenetic analysis of the curated NR family confirmed gene annotations and suggested that the MARs are a recent addition to the crustacean superfamily, occurring across the Malacostraca from the Stomatopoda to the Decapoda. The ligand-binding domain of these receptors appears to be less conserved than that of typical group-1 NRs. Expression data from two other crustacean species was utilized to examine MAR expression. The Y-organ of the tropical land crab showed a decline in expression of all MARs from intermolt to post-molt. Tissue distributions showed gonad-enriched expression in the Eastern rock lobster and antennal gland-enriched expression in the tropical land crab, although expression was evident across most tissues. CONCLUSION: By mining transcriptome data, we have curated an extensive list of NR genes expressed during the metamorphic molts of P. ornatus, including three novel crustacean NRs which appear to play a role in the molting process. Divergence of the E-region of these new receptors indicates that they may have adopted a function that is unconventional for NRs. Based on expression patterns, we can confirm that a number of NRs play a role in the ecdysone cassette which regulates molting in crustaceans. This study describes in detail the molecular events surrounding crustacean molting and metamorphosis by taking advantage of the distinctive life history unique to achelatan crustaceans.

Haemolymph microbiome of the cultured spiny lobster Panulirus ornatus at different temperatures.

Lobsters have an open circulatory system with haemolymph that contains microorganisms even in the healthy individuals. Understanding the role of these microorganisms becomes increasingly important particularly for the diagnosis of disease as the closed life-cycle aquaculture of the spiny lobster Panulirus ornatus nears commercial reality. This study aimed to characterise haemolymph responses of healthy cultured P. ornatus juveniles at control (28 °C) and elevated (34 °C) temperatures. This was assessed by measuring immune parameters (total granulocyte counts, total haemocyte counts, clotting times), and culture-independent (pyrosequencing of haemolymph DNA) and culture-dependent (isolation using nonselective growth medium) techniques to analyse bacterial communities from lobster haemolymph sampled on days 0, 4 and 6 post-exposure to the temperature regimes. Elevated temperature (34 °C) affected lobster survival, total granulocyte counts, and diversity, load and functional potential of the haemolymph bacterial community. Pyrosequencing analyses showed that the core haemolymph microbiome consisted of phyla Proteobacteria and Bacteriodetes. Overall, culture-independent methods captured a higher bacterial diversity and load when compared to culture-dependent methods, however members of the Rhodobacteraceae were strongly represented in both analyses. This is the first comprehensive study providing comparisons of haemolymph bacterial communities from healthy and thermally stressed cultured juvenile P. ornatus and has the potential to be used in health monitoring programs.

Aspectos técnicos e econômicos da engorda de lagostas: a experiência do vietnam e perspectiva de desenvolvimento da atividade no nordeste brasileiro / Technical and economic aspects of spiny lobster growout: the viatnamese experience and development perspective for the activity in northeastern Brazil / Aspectos técnicos y económicos del engorde de langosta: la experiencia de vietnan y perspectiva de desarrollo de la actividad en nordeste brasileño

A proposta desta revisão é resumir as informações sobre os estudos da engorda de lagostas em gaiolas no Vietnam. Esta sinopse demonstra que os juvenis podem ser cultivados sob regime de confinamento e se adapta bem as condições artificiais com alimentação. Na aquicultura, na engorda de juvenis de lagostas capturadas na natureza (Palinuridae) até o tamanho comercial está emergindo no Vietnam. No Vietnam o cultivo de lagostas em gaiolas no mar começou na década de 1990. Nesse contexto tradicionalmente a Panulirus ornatus e outras espécies são alimentadas com rejeito de pesca e pescado de baixo valor. O objetivo global da engorda pode ser o alívio da pobreza, e neste respeito para os criadores de lagostas é em particular uma oportunidade atrativa para brasileiros devido à captura de pós-larvas de lagostas (com autorização dos órgãos competentes) e sua engorda envolve tecnologia simples, mínimo de capital, e é apropriado como uma atividade alternativa de renda para pescadores.(AU)

The purpose of this review is to sumarize the studies on spiny lobster cage farming in Vietnam. The review showed that spiny lobster juveniles can be held in communal culture systems, since they adapt well to the artificial feeding conditions. In aquaculture, the growout phase of wild-caught juvenile spiny lobsters (Palinuridae) to market size is an emerging sector in Vietnam. Spiny lobster farming in sea cages commenced in the 1990s in Vietnam. Traditionally, Panulirus ornatus lobster and other species are fed with bycatch and cheap fish. The main purpose of the growout phase can be the decrease in poverty, and in this respect lobster farming is a particularly attractive opportunity for Brazil, since the capture of post-hatching lobsters (with authorization of the competent organs) and their growout involves simple technology, minimal capital and is ideally suited as an alternative activity for income generation for fisherman.(AU)

La propuesta de esta revisión es resumir las informaciones sobre los estudios de engorde de langostas en jaulas en Vietnan. Esta sinopsis demuestra que los juveniles pueden cultivarse en régimen de confinamiento y se adapta bien a las condiciones artificiales de alimentación. En la acuicultura, el engorde de juveniles de langostas capturadas en la naturaleza (Palinuridae) hasta el tamaño comercial, está emergiendo en Vietnan. Allá el cultivo de langostas en jaula en el mar empezó en la década de 1990. En ese contexto tradicionalmente la Panulirus ornatus y otras espécies son alimentadas con desechos de pesca y pescado de bajo valor. El objetivo global de engorde puede ser el alivio de la pobreza, y con relación a los creadores de langostas es en particular una atractiva oportunidade para brasileños, debido a la captura poslarvas de langostas (con autorización de los órganos competentes), y su engorde involucra tecnologia simple, mínimo de capital, y es apropriado como uma actividad alternativa de renta para los pescadores.(AU)

Can metamorphosis survival during larval development in spiny lobster Sagmariasus verreauxi be improved through quantitative genetic inheritance?

BACKGROUND: One of the major impediments to spiny lobster aquaculture is the high cost of hatchery production due to the long and complex larval cycle and poor survival during the many moult stages, especially at metamorphosis. We examined if the key trait of larval survival can be improved through selection by determining if genetic variance exists for this trait. Specifically, we report, for the first time, genetic parameters (heritability and correlations) for early survival rates recorded at five larval phases; early-phyllosoma stages (instars 1-6; S1), mid-phyllosoma stages (instars; 7-12; S2), late-phyllosoma stages (instars 13-17; S3), metamorphosis (S4) and puerulus stage (S5) in hatchery-reared spiny lobster Sagmariasus verreauxi. RESULTS: The data were collected from a total of 235,060 larvae produced from 18 sires and 30 dams over nine years (2006 to 2014). Parentage of the offspring and full-sib families was verified using ten microsatellite markers. Analysis of variance components showed that the estimates of heritability for all the five phases of larval survival obtained from linear mixed model were generally similar to those obtained from threshold logistic generalised models (0.03-0.47 vs. 0.01-0.50). The heritability estimates for survival traits recorded in the early larval phases (S1 and S2) were higher than those estimated in later phases (S3, S4 and S5). The existence of the additive genetic component in larval survival traits indicate that they could be improved through selection. Both phenotypic and genetic correlations among the five survival measures studied were moderate to high and positive. The genetic associations between successive rearing periods were stronger than those that are further apart. CONCLUSIONS: Our estimates of heritability and genetic correlations reported here in a spiny lobster species indicate that improvement in the early survival especially during metamorphosis can be achieved through genetic selection in this highly economic value species.

Developmental and gut-related changes to microbiomes of the cultured juvenile spiny lobster Panulirus ornatus.

With recent technologies making it possible for commercial scale closed life-cycle aquaculture production of spiny lobster (Panulirus ornatus) comes a strong impetus to further understand aspects of lobster health. The gut microbiome plays a crucial role in host health, affecting growth, digestion, immune responses and pathogen resistance. Herein we characterise and compare gut microbiomes across different developmental stages (6-7 days post-emergence [dpe], 52 dpe and 13 months post-emergence [mpe]) and gut regions (foregut, midgut and hindgut) of cultured P. ornatus juveniles. Gut samples were analysed using 16S rRNA next-generation sequencing. Core gut microbiomes of P. ornatus comprised the phyla Tenericutes and Proteobacteria. Within class Gammaproteobacteria, families Pseudoalteromonadaceae and Vibrionaceae were dominant members across the majority of the gut microbiomes. Characterisation of bacterial communities from 13 mpe lobsters indicated that the hindgut microbiome was more diverse and compositionally dissimilar to the foregut and midgut. The bacterial composition of the hindgut was more similar among younger juveniles (6-7 dpe and 52 dpe) compared to 13 mpe lobsters. This is the first study to explore gut microbiomes of spiny lobster juveniles. We demonstrate that the composition of the gut microbiome was shaped by gut region, whereas the structure of the hindgut microbiome was influenced by developmental stage.

CYP450s analysis across spiny lobster metamorphosis identifies a long sought missing link in crustacean development.

Cytochrome P450s (CYP450s) are a rapidly evolving family of enzymes, making it difficult to identify bona fide orthologs with notable lineage-specific exceptions. In ecdysozoans, a small number of the most conserved orthologs include enzymes which metabolize ecdysteroids. Ecdysone pathway components were recently shown in a decapod crustacean but with a notable absence of shade, which is important for converting ecdysone to its active form, 20-hydroxyecdysone (20HE), suggesting that another CYP450 performs a similar function in crustaceans. A CYPome temporal expression analysis throughout metamorphosis performed in this research highlights several un-annotated CYP450s displaying differential expression and provides information into expression patterns of annotated CYP450s. Using the expression patterns in the Eastern spiny lobster Sagmariasus verreauxi, followed by 3D modelling and finally activity assays in vitro, we were able to conclude that a group of CYP450s, conserved across decapod crustaceans, function as the insect shade. To emphasize the fact that these genes share the function with shade but are phylogenetically distinct, we name this enzyme system Shed.

Temperature dependent growth, feeding, nutritional condition and aerobic metabolism of juvenile spiny lobster, Sagmariasus verreauxi.

We examined the effects of temperature on the growth, feeding, nutritional condition and aerobic metabolism of juvenile spiny lobster, Sagmariasus verreauxi, in order to determine if temperature acclimated aerobic scope correlates with optimum for growth and to establish the thermal tolerance window for this emerging aquaculture species. Juvenile lobsters (initial weight=10.95±0.47g) were reared (n=7) at temperatures from 11.0 to 28.5°C for 145days. All lobsters survived from 14.5 to 25.0°C while survival was reduced at 11.0°C (86%) and all lobsters died at 28.5°C. Lobster specific growth rate and specific feed consumption displayed a unimodal response with temperature, peaking at 21.5°C. Lobster standard, routine and maximum metabolic rates, and aerobic scope all increased exponentially up to maximum non-lethal temperature. Optimum temperature for growth did not correspond to that for maximum aerobic scope suggesting that aerobic scope is not an effective predictor of the thermal optimum of spiny lobsters. Plateauing of specific feed consumption beyond 21.5°C suggests that temperature dependent growth of lobsters is limited by capacity to ingest or digest sufficient food to meet increasing maintenance metabolic demands at high temperatures. The nutritional condition of lobsters was not influenced by temperature and feed conversion ratio was improved at lower temperatures. These findings add to a growing body of evidence questioning the generality of aerobic scope to describe the physiological thermal boundaries of aquatic ectotherms and suggest that feed intake plays a crucial role in regulating performance at thermal extremes.

Applying the Power of Transcriptomics: Understanding Male Sexual Development in Decapod Crustacea.

The decapod Crustacea are the most species-rich order of the Crustacea and include some of the most charismatic and highly valued commercial species. Thus the decapods draw a significant research interest in relation to aquaculture, as well as gaining a broader understanding of these species' biology. However, the diverse physiology of the group considered with the lack of a model species have presented an obstacle for comparative analyses. In reflection of this, the recent integration of comparative transcriptomics has rapidly advanced our understanding of key regulatory pathways and developmental phenomena, an example being our understanding of sexual development. We discuss our work in the Eastern spiny lobster, Sagmariasus verreauxi, in the context of what is currently known about male sexual development in the decapods, highlighting the importance of transcriptomic techniques in achieving our recent advancements. We describe the progression made in our understanding of male sexual differentiation and maturation, as mediated by the insulin-like androgenic gland hormone (IAG), integrating the role of regulatory binding proteins (IGFBPs), a tyrosine kinase insulin receptor (TKIR), as well as the upstream effect of neuroendocrine hormones (GIH and MIH). We then consider the less well understood mechanism of male sex determination, with an emphasis on what we believe to be the key regulatory factors, the Dsx- and mab-3-related transcription factors (Dmrts). Finally, we discuss the function of the antennal gland (AnG) in sexual development, relating to the emergence of male-biased upregulation in the AnG in later sexual maturation and the sexually dimorphic expression of two key genes Sv-TKIR and Sv-Dmrt1 We then present the AnG as a case study to illustrate how comparative transcriptomic techniques can be applied to guide preliminary analyses, like the hypothesis that the AnG may function in pheromone biosynthesis. In summary, we describe the power of transcriptomics in facilitating the progress made in our understanding of male sexual development, as illustrated by the commercial decapod species, S. verreauxi Considering future directions, we suggest that the integration of multiple omics-based techniques offers the most powerful tool to ensure we continue to piece together the biology of the important group of decapod Crustacea.

Seismic air gun exposure during early-stage embryonic development does not negatively affect spiny lobster Jasus edwardsii larvae (Decapoda: Palinuridae).

Marine seismic surveys are used to explore for sub-seafloor oil and gas deposits. These surveys are conducted using air guns, which release compressed air to create intense sound impulses, which are repeated around every 8-12 seconds and can travel large distances in the water column. Considering the ubiquitous worldwide distribution of seismic surveys, the potential impact of exposure on marine invertebrates is poorly understood. In this study, egg-bearing female spiny lobsters (Jasus edwardsii) were exposed to signals from three air gun configurations, all of which exceeded sound exposure levels (SEL) of 185 dB re 1 µPa(2) · s. Lobsters were maintained until their eggs hatched and the larvae were then counted for fecundity, assessed for abnormal morphology using measurements of larval length and width, tested for larval competency using an established activity test and measured for energy content. Overall there were no differences in the quantity or quality of hatched larvae, indicating that the condition and development of spiny lobster embryos were not adversely affected by air gun exposure. These results suggest that embryonic spiny lobster are resilient to air gun signals and highlight the caution necessary in extrapolating results from the laboratory to real world scenarios or across life history stages.

Inter-cohort growth for three tropical resources: tilapia, octopus and lobster.

Growth parameters are an important component for the stock assessment of exploited aquatic species. However, it is often difficult to apply direct methods to estimate growth and to analyse the differences between males and females, particularly in tropical areas. The objective of this study was to analyse the inter-cohort growth of three tropical resources and discuss the possible fisheries management implications. A simple method was used to compare individual growth curves obtained from length frequency distribution analysis, illustrated by case studies of three tropical species from different aquatic environments: tilapia (Oreochromis aureus), red octopus (Octopus maya) and the Caribbean spiny lobster (Panulirus argus). The analysis undertaken compared the size distribution of males and females of a given cohort through modal progression analysis. The technique used proved to be useful for highlighting the differences in growth between females and males of a specific cohort. The potential effect of extrinsic and intrinsic factors on the organism's development as reflected in the size distribution of the cohorts is discussed.

Inter-cohort growth for three tropical resources: tilapia, octopus and lobster / Crecimiento ínter-cohorte para tres recursos tropicales: tilapia, pulpo y langosta

Growth parameters are an important component for the stock assessment of exploited aquatic species. However, it is often difficult to apply direct methods to estimate growth and to analyse the differences between males and females, particularly in tropical areas. The objective of this study was to analyse the inter-cohort growth of three tropical resources and discuss the possible fisheries management implications. A simple method was used to compare individual growth curves obtained from length frequency distribution analysis, illustrated by case studies of three tropical species from different aquatic environments: tilapia (Oreochromis aureus),red octopus (Octopus maya)and the Caribbean spiny lobster (Panulirus argus).The analysis undertaken compared the size distribution of males and females of a given cohort through modal progression analysis. The technique used proved to be useful for highlighting the differences in growth between females and males of a specific cohort. The potential effect of extrinsic and intrinsic factors on the organism's development as reflected in the size distribution of the cohorts is discussed.

Los parámetros de crecimiento son un componente importante para la evaluación de las poblaciones de especies acuáticas explotadas. Sin embargo, es complicado aplicar métodos directos para estimar el crecimiento y analizar las diferencias entre machos y hembras particularmente en zonas tropicales. El objetivo de este estudio fue analizar el crecimiento entre cohortes de tres recursos tropicales y discutir las posibles implicaciones en el manejo pesquero. Se utilizó un método simple para comparar las curvas de crecimiento individual obtenidas a través del análisis de distribución de frecuencias de longitudes, para ello se usaron tres casos de estudio de especies tropicales provenientes de diferentes ambientes acuáticos: tilapia (Oreochromis aureus),pulpo rojo (Octopus maya)y la langosta del caribe (Panulirus argus).El análisis consistió en comparar la distribución de longitudes de hembras y machos obtenidas del análisis de progresión modal. Esta técnica puede ser utilizada para resaltar las diferencias en el crecimiento entre hembras y machos de una cohorte específica. En este documento se discute el efecto potencial de los factores intrínsecos y extrínsecos en el desarrollo de los organismos como se refleja en la distribución de tallas de las cohortes.

Strengthened currents override the effect of warming on lobster larval dispersal and survival.

Human-induced climate change is projected to increase ocean temperature and modify circulation patterns, with potential widespread implications for the transport and survival of planktonic larvae of marine organisms. Circulation affects the dispersal of larvae, whereas temperature impacts larval development and survival. However, the combined effect of changes in circulation and temperature on larval dispersal and survival has rarely been studied in a future climate scenario. Such understanding is crucial to predict future species distributions, anticipate ecosystem shifts and design effective management strategies. We simulate contemporary (1990s) and future (2060s) dispersal of lobster larvae using an eddy-resolving ocean model in south-eastern Australia, a region of rapid ocean warming. Here we show that the effects of changes in circulation and temperature can counter each other: ocean warming favours the survival of lobster larvae, whereas a strengthened western boundary current diminishes the supply of larvae to the coast by restricting cross-current larval dispersal. Furthermore, we find that changes in circulation have a stronger effect on connectivity patterns of lobster larvae along south-eastern Australia than ocean warming in the future climate so that the supply of larvae to the coast reduces by ~4% and the settlement peak shifts poleward by ~270 km in the model simulation. Thus, ocean circulation may be one of the dominant factors contributing to climate-induced changes of species ranges.

Bioenergetics of Nutrient Reserves and Metabolism in Spiny Lobster Juveniles Sagmariasus verreauxi: Predicting Nutritional Condition from Hemolymph Biochemistry.

The nutritional condition of cultured Sagmariasus verreauxi juveniles over the molt and during starvation was investigated by studying their metabolism, bioenergetics of nutrient reserves, and hemolymph biochemistry. Juveniles were shown to downregulate standard metabolic rate by as much as 52% within 14 d during starvation. Hepatopancreas (HP) lipid was prioritized as a source of energy, but this reserve represented only between 1% and 13% of the total measured energy reserve and was used quickly during starvation, especially in the immediate postmolt period when as much as 60% was depleted within 3 d. Abdominal muscle (AM) protein represented between 74% and 90% of the total measured energy reserve in juvenile lobsters, and as much as 40% of available AM protein energy was used over 28 d of starvation after the molt. Carbohydrate reserves represented less than 2% of the measured total energy reserve in fed intermolt lobsters and provided negligible energy during starvation. Eighteen hemolymph parameters were measured to identify a nondestructive biomarker of condition that would reflect accurately the state of energy reserves of the lobster. Among these, the hemolymph Brix index was the most accurate and practical method to predict HP lipid and the total energy content of both the HP and the AM in juvenile S. verreauxi. The Brix index was strongly correlated with hemolymph proteins, triglyceride, cholesterol, calcium, and phosphorus concentrations, as well as lipase activity; all were useful in predicting condition. Electrolytes such as chloride, magnesium, and potassium and metabolites such as glucose and lactate were poor indicators of nutritional condition. Uric acid and the "albumin"-to-"globulin" ratio provided complementary information to the Brix index, which may assist in determining nutritional condition of wild juvenile lobsters of unknown intermolt development. This study will greatly assist future ecological studies examining the nutritional condition of juvenile lobsters in the wild, as well as the development of husbandry protocols and feeds for aquaculture.

Inducers of settlement and moulting in post-larval spiny lobster.

The rapid and often remote location of suitable habitats used by migrating organisms is often critical to their subsequent recruitment, fitness and survival, and this includes in the marine environment. However, for the non-feeding post-larval stage of spiny lobsters, effective settlement cues for habitat selection are critical to their success but are poorly described. Therefore, the current study examined whether acoustic and substrate cues have the potential to shorten the time to moulting and affect their subsequent nutritional condition in the pueruli of the southern spiny lobster, Jasus edwardsii. Individuals moulted to first instar juveniles up to 38% faster when exposed to the underwater sound from two types of typical settlement habitat (coastal kelp- and urchin-dominated reefs) compared to those with no underwater sound. The settlement delay in the post-larvae without underwater sound also resulted in juveniles in poorer survival and nutritional condition as measured by their protein and lipid contents. In a separate experiment, post-larvae presented with seaweed and rock substrates were found to complete settlement and moult to juvenile by as much as 20% faster compared to those on the sand and control treatments. Overall, the results are the first to demonstrate that the pueruli of J. edwardsii have the ability to detect and respond to underwater sound, as well as determining that both acoustic and substrate cues play a role in modulating physiological development during settlement.

Discovery of a novel insulin-like peptide and insulin binding proteins in the Eastern rock lobster Sagmariasus verreauxi.

This study reports, for the first time in any of the commercially important decapod species, the identification of an insulin-like peptide (ILP), distinct from the androgenic gland hormone. Bioinformatics analysis of the de novo assembled spiny lobster, (Sagmariasus verreauxi) transcriptome, allowed identification of Sv-ILP1 as well as eight binding proteins. Binding proteins were termed as Sv-IGFBP, due to homology with the vertebrate insulin-like growth-factor binding protein and Sv-SIBD1-7, single insulin-binding domain protein (SIBD), similar to those identified in other invertebrate species. Sv-ILP1 was found to be expressed in the eyestalk, gonads and antennal gland of both sexes and to a lesser extent in male muscle, androgenic gland and hepatopancreas. The expression profiles of each binding protein were found to vary across tissues, with Sv-SIBD5, 6 and 7 showing higher expression in the gonad, demonstrated by PCR and digital gene expression. Further spatial investigations, using in-situ hybridisation, found Sv-ILP1 to be expressed in the neurosecretory cells of the thoracic ganglia, in keeping with the tissue expression of Drosophila ILP7 (DILP7). This correlative tissue expression, considered with the phylogenetic clustering of Sv-ILP1 and DILP7, suggests Sv-ILP1 to be a DILP7 orthologue. The broad expression of Sv-ILP1 strongly suggests that ILPs have a role beyond that of masculinisation in decapods. The function of these novel peptides may have application in enhancing aquaculture practices in the commercially important decapod species.

Identification and characterization of androgenic gland specific insulin-like peptide-encoding transcripts in two spiny lobster species: Sagmariasus verreauxi and Jasus edwardsii.

In this study we describe, for the first time in spiny lobsters, the androgenic gland and its putative hormone. The androgenic gland in crustaceans is the key regulator of crustacean masculinity. The transcript encoding the insulin-like androgenic gland specific factor has recently been identified and characterized in a number of decapod crustacean species including commercially important crabs, crayfish, prawns and shrimps. This insulin-like factor has proven to be the androgenic gland masculinizing hormone, and is absent in females. While the androgenic gland and its putative hormone have been identified in all other commercially valuable groups, none had been identified in lobsters. We identified and characterized the androgenic glands of two spiny lobster species (Sagmariasus verreauxi and Jasus edwardsii) and conducted a transcriptomic analysis of the S. verreauxi androgenic gland. Bioinformatics analysis led to the discovery and characterization of the insulin-like androgenic gland specific factors in both species studied. Changes in androgenic gland cell size and quantity between sub-adult and sexually mature males were evident. The transcriptomic database established for the S. verreauxi androgenic gland might enable to elucidate the mechanisms through which the insulin-like factor is secreted, transported to the target cells and how it triggers the physiological effects of sexual differentiation towards maleness and maintenance of the male gonad.

Male Sexual Development and the Androgenic Gland: Novel Insights through the de novo Assembled Transcriptome of the Eastern Spiny Lobster, Sagmariasus verreauxi.

The Eastern spiny lobster, Sagmariasus verreauxi, is commercially important in fisheries, with growing aquaculture potential, driving an interest to better understand male sexual differentiation. Amongst the Decapoda, the androgenic gland (AG) and the insulin-like androgenic gland hormone (IAG) have a well-defined function in male sexual differentiation. However, IAG is not a sex determinant and therefore must be considered as part of a broader, integrated pathway. This work uses a transcriptomic, multi-tissue approach to provide an integrated description of male-biased expression as mediated through the AG. Transcriptomic analyses demonstrate that IAG expression is stage- and eyestalk-regulated (low in immature, high in mature and 6-times higher in hypertrophied glands), with IAG being the predominant AG-specific factor. The low expression of this key factor in immature males suggests the involvement of other tissues in male sexual differentiation. Across tissues, the gonad (87.8%) and antennal gland (73.5%) show the highest male-biased differential expression of transcripts and also express 4 sex-determination regulators, known as Dmrts, with broader expression of Sv-Sxl and Sv-TRA-2. In order to better understand male sexual differentiation, tissues other than the AG must also be considered. This research highlights the gonad and antennal gland as showing significant male-biased expression patterns, including the Sv-Dmrts.