Search

The West Coast rock lobster (WCRL), Jasus lalandii, inhabits highly variable environments frequented by upwelling events, episodes of hypercapnia and large temperature variations. Coupled with the predicted threat of ocean acidification and temperature change for the coming centuries, the immune response in this crustacean will most likely be affected. We therefore tested the hypothesis that chronic exposure to hypercapnia and elevated seawater temperature will alter immune function of the WCRL. The chronic effects of four combinations of two stressors (seawater pCO2 and temperature) on the total number of circulating haemocytes (THC) as well as on the lobsters’ ability to clear (inactivate) an injected dose of Vibrio anguillarum from haemolymph circulation were assessed. Juvenile lobsters were held in normocapnic (pH 8.01) or hypercapnic (pH 7.34) conditions at two temperatures (15.6 and 18.9 °C) for 48 weeks (n = 30 lobster per treatment), after which a subsample of lobsters (n = 8/treatment), all at a similar moult stage, were selected from each treatment for the immune challenge. Baseline levels of haemocytes (THC ml−1) and bacteria (CFU ml−1) in their haemolymph were quantified 24 h prior to bacterial challenge. Lobsters were then challenged by injecting 4 × 104 V. anguillarum per g body weight directly into the cardiac region of each lobster and circulating haemocyte and culturable bacteria were measured at 20 min post challenge. No significant differences in THC ml−1 (p normocapnia/high temperature > normocapnia/low temperature > hypercapnia/high temperature. This study demonstrated that despite chronic exposure to combinations of reduced seawater pH and high temperature, the WCRL was still capable of rapidly rendering an injected dose of bacteria non-culturable.

Ammonia is a common pollutant in the aquatic ecosystem and closed aquaculture systems. It may pose a threat to the lobster growth, reproduction and survival. However, there is lack of research of the mechanisms on the toxic effects ammonia at molecular levels. In this work, proteomics and metabolomics were integrated to analyze the proteome and metabolome responses in the ornate spiny lobster Panulirus ornatus treated with ammonia (20 mg L

Anthropogenic aquatic noise is recognised as an environmental pollutant with the potential to negatively affect marine organisms. Seismic surveys, used to explore subseafloor oil reserves, are a common source of aquatic noise that have garnered attention due to their intense low frequency inputs and their frequent spatial overlap with coastal fisheries. Commercially important Southern Rock Lobster (Jasus edwardsii) adults have previously shown sensitivity to signals from a single seismic air gun. Here, the sensitivity of J. edwardsii juveniles and puerulus to the signals of a full-scale seismic survey were evaluated to determine if early developmental stages were affected similarly to adults, and the range of impact. To quantify impact, lobster mortality rates, dorsoventral righting reflex and progression through moult cycle were evaluated following exposure. Exposure did not result in mortality in either developmental stage, however, air gun signals caused righting impairment to at least 500 m in lobsters sampled immediately following exposure, as had previously been reported in adults with corresponding sensory system damage following exposure. Impairment resulting from close range (0 m) exposure appeared to be persistent, as previously reported in adults, whereas juveniles exposed at a more distant range (500 m) showed recovery, indicating that exposure at a range of 500 m may not cause lasting impairment to righting. Intermoult duration was (time between moults) significantly increased in juveniles exposed at 0 m from the source, indicating the potential for slowed development, growth, and physiological stress. These results demonstrate that exposure to seismic air gun signals have the potential to negatively impact early life history stages of Southern Rock Lobsters. The similarity of both the impacts and the sound exposure levels observed here compared to previous exposure using a single air gun offer validation for the approach, which opens the potential for accessible field-based experimental work into the impact of seismic surveys on marine invertebrates.

Lobsters are fished world-wide due to their status as a high value, luxury seafood. A large proportion of the product is sold via live export, with lobsters subject to a range of stressors during holding post-capture. Improving the current understanding of the immune response to these stressors assists in improving efficiency and reducing loss in the chain between capture and consumption. In this study, the immune status of four treatment groups of Southern Rock Lobster (Jasus edwardsii) were studied: controls recently landed from a fishing boat, lobsters displaying advanced shell necrosis, lobsters in an unexplained moribund state and lobsters held in a processing facility for 10 weeks in standard conditions (i.e. high density, fasted). A total of 15 immune parameters and 19 haemolymph biochemical parameters were assayed. Phenoloxidase activity was only sporadically observed in haemocyte lysate and was consistently observed at a low level in the plasma with no difference between treatments for either. Haemocyte lysate prophenoloxidase activity was detected in most individuals, with no differences found between treatments. Prophenoloxidase in the plasma showed the highest level of activity, with the shell necrosis treatment demonstrating an elevated activity level relative to the other three treatments. Cell viability was not affected in any treatment. Lobsters with shell necrosis had a reduced capacity for phagocytosis, a significantly higher total haemocyte count, fewer hyalinocytes and more granulocytes and semigranulocytes. Fasted lobsters showed an opposite shift, with significantly more hyalinocytes compared to the other treatments and very few granulocytes and semigranulocytes. The balance of a range electrolytes, minerals metabolites and enzymes were affected in shell necrosis and fasted treatments, raising them as potential markers for immunocompromised lobsters. Multivariate analysis of all assayed parameters showed that all individuals in the necrosis treatment showed a similar, distinct immune response and that the fasted treatment, along with one control and one moribund individual, showed a separate intermediate response. The remainder of the control and moribund lobsters demonstrated a distinct "non-response" in comparison. These results offer a characterisation of the physiological response to common challenges during post-capture holding of rock lobsters, demonstrating the differential response to pathogenic bacterial infection, long term fasting, non-specific moribundity and the stress of capture and transport.

Crustaceans suffer from diseases that can alter their survival and ecology with additional economic consequences for fisheries and aquaculture. Many parasites have been described from crustaceans and with the advent of novel technologies such as next generation sequencing, the discovery of novel parasites has become increasingly efficient. Molecular techniques are beginning to surpass more conventional tools for parasite discovery, but they typically do not provide information on pathology. Histopathology remains one of the least expensive methods for parasite discovery and allows for both detection of parasites and descriptions of the pathology they cause. When used in concert with modern molecular and electron microscopy techniques, the approach is powerful; however, there are few informational tools for the interpretation of histological slides from crustaceans. Those available do not provide comprehensive images of all organs and early works were limited to lower resolution than currently available. More recent texts provide in-depth details of infection in histological section, but few provide images of healthy material or describe a baseline from which to compare. Here, we provide a series of image plates derived from histologically processed tissues from three palinurid lobsters: Panulirus argus, Palinurus elephas and Panulirus guttatus. Histology from these lobsters shows high visual similarity in all tissue types. We provide a histological atlas of healthy tissue that can be used as a baseline resource for pathobiologists working on these common species (and related crustaceans) and we discuss how disease may result in visual aberrations to these tissues.

The Caribbean spiny lobster Panulirus argus is susceptible to infection by Panulirus argus Virus 1 (PaV1), the only virus known to naturally infect any lobster species. However, P. argus is able to mitigate PaV1 transmission risk by avoiding infected individuals. White Spot Syndrome Virus (WSSV) has a particularly wide host range. WSSV has not been documented in wild populations of spiny lobsters, but has been experimentally transmitted to six other lobster species from the genus Panulirus spp. While WSSV has been detected intermittently in wild populations of shrimp in the Caribbean region, the risk to P. argus has not been evaluated. Potential emergence of the disease could result in fisheries losses and ecological disruption. To assess the risk to P. argus, we tested its susceptibility to WSSV via injection and waterborne transmission. We also tested whether healthy lobsters can detect and avoid conspecifics with qPCR-quantifiable WSSV infections. We found P. argus to be highly susceptible to WSSV via intramuscular injection, with mortality reaching 88% four weeks post inoculation. Panulirus argus was also susceptible to WSSV via waterborne transmission, but WSSV burden was low after four weeks via qPCR. Behavioral assays indicated that P. argus can detect and avoid conspecifics infected with WSSV and the avoidance response was strongest for the most heavily infected individuals – a response comparable to PaV1-infected conspecifics. Panulirus argus is the first spiny lobster found to be susceptible to WSSV in the Americas, but it is possible that a generalized avoidance response by healthy lobsters against infected conspecifics provides a behavioral defense and may reduce WSSV infection potential and prevalence. Such avoidance may extend to other directly transmitted pathogens in spiny lobster populations preventing them from becoming common in their population.Author SummaryErica P. Ross is a PhD candidate at the University of Florida, studying the disease ecology of the Caribbean spiny lobster, with a focus on chemosensory ecology. Donald C. Behringer is an associate professor at the University of Florida and his research focuses on disease ecology, epidemiology, and fishery ecology, with a focus on crustaceans and other marine invertebrates. Jamie Bojko received his PhD from the University of Leeds and is currently a post-doctorate associate at the University of Florida studying experimental and systemic crustacean pathology.

The Caribbean spiny lobster, Panulirus argus (Latreille, 1804) supports important fisheries in the Caribbean region. This species is affected by a deadly virus, Panulirus argus Virus 1 (PaV1), the only known pathogenic virus for this species. As infection progresses, the effects of PaV1 on its host become systemic, with far reaching impacts on the host's physiology, including structural injuries to its gastrointestinal organs, such as the hepatopancreas and the gut. This last one becomes highly compromised in the last stages of infection. Since the gut is a key organ for the physiological stability of lobsters, we compared the transcriptomic changes in the gut of juvenile individuals of Panulirus argus naturally infected with PaV1. In the RNA-Seq analysis, we obtained a total of 485 × 10

Phenoloxidases (POs) play a crucial role in melanization of crustaceans. There are at least two types of POs characterized in crustaceans: the conventional type (POα here) that is expressed in hemocytes and POβ, a secreted protein synthesized in the hepatopancreas. We investigated the source of PO activity in the hemolymph of a lobster and determined the kinetic parameters of mono- and di-PO activities. In the lobster hemolymph, POα, which formed a hexamer similar to both POβ and hemocyanin, contributed to PO activity, whereas the amount of POβ was low. Kinetic analyses using purified prophenoloxidase of crustaceans showed that lobster POα has a higher rate constant, while shrimp POβ has higher specificity in both mono- and di-PO reactions, when tyramine and dopamine were employed as substrates. There should be at least two types of PO molecules in crustacean hemolymph, but the dominant PO molecule type varies among species.

While the California spiny lobster (Panulirus interruptus) is an important commercial and recreational fishery species in California, there is a lack of data on bioaccumulation for the species. This study examined pollutant tissue concentrations in lobsters from San Diego Bay, California. Observed lobster pollutant tissue concentrations in tail muscle were compared to State of California pollutant advisory levels. Concentrations were then used to conduct risk assessment using catch data from the California Department of Fish and Wildlife. Study results found little bioaccumulation of organic pollutants in tail tissue, likely due to low observed lipids. Mercury was present, predominantly in methyl form, at concentrations above advisory levels. Recreational catch data for San Diego Bay showed increased non-cancer risk for fishers at the 90th percentile or greater of reported annual catch. Further studies should focus on non-tail tissues, as exploratory whole lobster samples (n = 2) showed elevated organic pollutants and metals.

There is a critical knowledge gap regarding the impacts of seismic air gun signals on the physiology of adult crustaceans. We conducted four controlled field experiments to examine the impact of seismic acoustic signals on spiny lobster, Jasus edwardsii. Seismic air gun exposure suppressed total haemocyte count (THC) for up to 120 days post-exposure, suggesting a chronic negative impact of immune competency. THC levels after 365 days post-exposure, were elevated two fold, potentially indicating an immune response to infection. Haemolymph refractive index was reduced after 120 days post exposure in one experiment, suggesting a chronic impairment of nutritional condition. There was no effect of air gun exposure on 24 haemolymph biochemical parameters, hepatopancreas index or survival. Collectively these results indicate that the biochemical haematological homeostasis of J. edwardsii is reasonably resilient to seismic acoustic signals, however, air gun exposure may negatively influence the lobster's nutritional condition and immunological capacity.

The literature on late-stage larval and puerulus settlement of tropical spiny lobsters suggests that a broad range of visually-assessed variables may play a role in the selection of settlement substrates. These variables include light level, luminosity, degree of exposure, and the availability of edges, crevices and clefts. A fishery has developed in Indonesia, that captures pueruli for grow-out purposes, along the entire southern coastline from Java to Sumbawa. The present study builds on knowledge gained on the behaviour and substrate preferences of settling pueruli by examining aspects of collector design and substrate surface characteristics in regard to catch rates. A series of tank-based experiments were used to examine preferences for various attributes of collector materials, and preferred material characteristics were then tested in a field experiment to verify the laboratory results. In the tank experiment the most preferred substrate was cement bag paper, followed by insect mesh, weed netting, PVC rubber and cement bag plastic. Crevice angles of 10° and 20° were significantly preferred over 30°. The substrate level and other substrates had less influence on settlement. However, both the lunar phase and type of materials tested in the field had a significant effect on settlement. Significantly greater numbers of pueruli were caught at the time of the new moon than at other moon phases. Cement bag paper was the most preferred material in the field, followed by cement bag plastic, insect mesh, weed fabric, and PVC rubber.

Sex determination pathways are extensively diverse across species, with the master sex-determinants being the most variable element. Despite this, there is a family of DM-domain transcription factors (Dmrts), which hold a highly conserved function in sexual development. This work is the first to describe a heterogametic sex-linked Dmrt in an invertebrate species, the Eastern spiny lobster, Sagmariasus verreauxi. We have termed the Ylinked, truncated paralogue of the autosomal iDmrt1, Sv-iDMY. Considering the master sex-determining function of both DMY in medaka and DM-W in frog, we hypothesised a similar function of Sv-iDMY. By conducting temporal expression analyses during embryogenesis we have identified a putative male sexdetermining period during which iDMY > iDmrt1. Employing a GAL4-transactivation assay we then demonstrate the dominant negative suppression of iDMY over its autosomal iDmrt1 paralogue, suggesting the mechanism with which iDMY determines sex. Comparative analyses of Sv-iDMY, DM-W and medaka DMY, highlight the C′-mediated features of oligomerisation and transactivation as central to the mechanism that each exerts. Indeed, these features may underpin the plasticity facilitating the convergent emergence of these three sporadic sex-linked master-Dmrts.

Fishery landings composition in tropical developing countries is formed of high diversity of species and some of them are not well-reported in landings or national fisheries statistics. This is particularly the case of the endemic red spiny lobster Panulirus penicillatus (Olivier, 1791) on the Pacific coast of Costa Rica. The red spiny lobster is a circumtropical species which has the most extensive global distribution of all the species of spiny lobsters. In order to fill the gap of knowledge regarding the spatial distribution of this species in the eastern Pacific Ocean and highlight its importance to fishing communities, this study focused on data collected from a dive-based bentho-demersal fishery on the northern Pacific coast of Costa Rica. Environmental and fishing effort data were recorded as an input for fitting site-occupancy models to get estimates of occurrence and probability of detection. The most parsimonious model structure for detection suggested that the covariates of diving time and water column visibility had an important effect on detection probability. For the occupancy part of the model, the depth gradient had the strongest effect, with high probability of lobster encounters in shallow rocky reefs near the coastline. These findings point towards an enlargement of the area of occurrence of the endemic sub-population of P. penicillatus to which sampled animals belong. Further, it is discussed potential ecological and fishery factors that explain the low occurrence of the red spiny lobster at this restricted area of the eastern Pacific continental shelf. As a result, a set of research guidelines and management recommendations are suggested for promoting the conservation and sustainable use of this endemic species within the region.

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.

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 ± 0.47 g) were reared (n = 7) at temperatures from 11.0 to 28.5 °C for 145 days. 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.

Beta_defensin have been solely found in vertebrates until β-defensin-like peptides were described as transcript isoforms in two species of Panulirus genus. They were considered as putative antimicrobials since their biological activity have not been demonstrated. Here we purified and characterized a defensin-like peptide from the hemocytes of spiny lobster P. argus, hereafter named panusin. Structurally, panusin presents a cysteine-stabilized α/β motif, and is prone to form homodimers. Biological activity of panusin showed broad-spectrum antimicrobial activity, characterized for being strikingly salt-resistant. Panusin did not showed hemolytic activity but was demonstrated its binding capacity to different lipid membrane models, indicating amphipathicity of β-sheet core as driving force for its antimicrobial activity. Panusin is considered a new kind of arthropod defensin which share structural and biological features with beta-defensin from vertebrates. The presence of beta-defensin like peptides in crustacean might suggest the emergence of the evolutionary relationship of β-defensins from vertebrates., This work was partially supported by IFS Grant reference No F/5024-1 and F/5024-2, and by the VLIR-UOS Grant reference ZEIN2013Z134

Mortality events during live transport of Jasus edwardsii rock lobsters are common around the time of season openings in Tasmania, with lobsters from deeper fishing areas with pale shell colouration (brindle) being perceived as more susceptible than shallow-water, red-coloured (red) lobsters. The aims of this study were to assess and predict the vulnerability of brindle and red lobsters to extended emersion exposure using pre- and post-emersion data which included 28 haemolymph biochemical parameters and 5 behaviour traits. No effect of lobster shell colour on haemolymph biochemistry, behaviour traits and their vulnerability to emersion was found. A combined survival of 97% after 40h and 57% after 64h in a first experiment, and 37% after 64h in a second experiment, was observed. Behaviour traits (i.e., righting response, tail flips and three reflex behaviours) were poor indicator of survival. Haemolymph parameters were either unaffected by emersion (e.g., Brix index, protein and lipids), affected by emersion but not associated with mortality (e.g., total haemocyte counts, calcium, magnesium, bicarbonate, glucose and uric acid), or associated with mortality following a recovery period (e.g., pH, the sodium to potassium ratio, urea, and the activity of amylase). A build-up of anaerobic end-products and nitrogenous waste most likely resulted in the mortality. A model based on lobster size and the pre-emersion concentration of haemolymph bicarbonate and haemocyanin was found to be a useful indicator of future survival. This study provides promising leads towards the development of a blood based vulnerability test for live crustacean prior transport.

Staotcysts, the mechanosensory organs common to many marine invertebrates, have shown sensitivity to aquatic noise. Previously, rock lobsters (Jasus edwardsii) from a remote site with little exposure to anthropogenic noise incurred persistent damage to the statocyst and righting reflex following exposure to seismic air gun signals. Here, J. edwardsii collected from a site subject to high levels of anthropogenic noise were exposed to an equivalent seismic air gun signal regime as the previous study of noise-naïve lobsters. Following exposure, both control and exposed treatments were found to have damage to the statocyst equivalent to that of noise-naïve lobsters following seismic exposure, which led to the conclusion that the damage was pre-existing and not exacerbated by seismic exposure. The source of the damage in the lobsters in this study could not be ascertained, but the soundscape comparisons of the collection sites showed that the noisy site had a 5-10 dB greater level of noise, equivalent to a 3-10 times greater intensity, in the 10-700 Hz range than was found at the remote collection site. In addition to the lack of further damage following seismic exposure, no disruption to the righting reflex was observed. Indeed, compared to the noise naïve lobsters, the lobsters here demonstrated an ability to cope with or adapt to the mechanosensory damage, indicating a need for better understanding of the ecological impacts of the damage caused by low frequency noise on marine organisms. More broadly, this study raises historical exposure to noise as a previously unrecognised but vitally important consideration for studies of aquatic noise.

Indonesia has a unique opportunity to establish the world's largest lobster aquaculture industry, based on a significant natural resource of settling puerulus. These seed lobsters can be captured and on-grown to generate high value, consumption size lobsters. However, existing fisheries regulation prohibits fishing of these seed lobsters in Indonesia. Nevertheless, seed fishing occurs widely, and the seed are smuggled into Vietnam, where the Vietnamese gain most of the commercial benefit. By removing access to pueruli and grow-out opportunities, these regulations have diminished the capacity of Indonesian lobster farmers to develop a sustainable grow-out industry. Although the new policies were introduced as a management intervention to protect adult lobster stocks, they have had the unintended consequence of preventing development of lobster aquaculture while having no known benefit on abundance of adult lobsters. This study collected data on the puerulus available in Indonesia, species composition, seasonality, and capture methods. A census of seed capture determined that 5,243,887 pueruli were fished from two bays in the southeast of Lombok in 2014, representing 64,787 pueruli per km2. Surveys beyond Lombok determined there was high abundance of seed in areas with similar environmental characteristics, where local communities had also engaged in seed fishing. The locations identified as having substantial puerulus abundance comprised the southern coast of Java, Bali, Lombok and Sumbawa covering a distance of 1500 km. Puerulus abundance is highest in enclosed bays with distinct currents, relatively high turbidity attributable to terrestrial inflows, and muddy/sandy substrates. Satellite imaging identified all such suitable locations for puerulus settlement, enabling an estimate of total puerulus catch of 103,480,283 per year. The composition of puerulus fished consisted of two species, Panulirus homarus and P. ornatus. P. homarus was most abundant representing between 63 and 87%, while P. ornatus represented 37 to 13%. Seasonality of catch was unclear, with low catch between December and March and higher catches from April to November. The magnitude of the estimated puerulus resource of Indonesia is >20 times greater than that of Vietnam. If used for aquaculture, the seed available can support an industry generating >12,500 t of market size lobster, and provide social and economic benefit to many thousands of households. To achieve this, fisheries policies could be revised and a regulatory framework established to manage the puerulus fishery and support lobster farming. Tropical marine rock lobster aquaculture is a major opportunity for high-value aquaculture production for the Indonesian economy.

The spiny lobster Panulirus argus (Latreille, 1804) is currently affected by an unenveloped, icosahedral, DNA virus termed Panulirus argus virus 1 (PaV1), a virulent and pathogenic virus that produces a long-lasting infection that alters the physiology and behaviour of heavily infected lobsters. Gut-associated microbiota is crucial for lobster homeostasis and well-being, but pathogens could change microbiota composition affecting its function. In PaV1 infection, the changes of gut-associated microbiota are yet to be elucidated. In the present study, we used high-throughput 16S rRNA sequencing technology to compare the bacterial microbiota in intestines of healthy and heavily PaV1-infected male and female juveniles of spiny lobsters P. argus captured in Puerto Morelos Reef lagoon, Quintana Roo, Mexico. We found that basal gut-associated microbiota composition showed a sex-dependent bias, with females being enriched in amplicon sequence variants (ASVs) assigned to Sphingomonas, while males were enriched in the genus Candidatus Hepatoplasma and Aliiroseovarius genera. Moreover, the alpha diversity of microbiota decreased in PaV1-infected lobsters. A significant increase of the genus Candidatus Bacilloplasma was observed in infected lobsters, as well as a significant decrease in Nesterenkonia, Caldalkalibacillus, Pseudomonas, Cetobacterium and Phyllobacterium. We also observed an alteration in the abundances of Vibrio species. Results from this study suggest that PaV1 infection impacts intestinal microbiota composition in Panulirus argus in a sex-dependent manner.

In crustaceans the insulin-like androgenic gland hormone (IAG) is responsible for male sexual differentiation. To date, the biochemical pathways through which IAG exerts its effects are poorly understood and could be elucidated through the production of a functional recombinant IAG (rIAG). We have successfully expressed glycosylated, biologically active IAG using the Pichia pastoris yeast expression system. We co-expressed recombinant single-chain precursor molecules consisting of the B and A chains (the mature hormone) tethered by a flexible linker, producing rIAGs of the following commercially important species: Eastern spiny lobster Sagmariasus verreauxi (Sv), redclaw crayfish Cherax quadricarinatus (Cq) and giant freshwater prawn Macrobrachium rosenbergii (Mr). We then tested the biological activity of each, through the ability to increase phosphorylation in the testis; both Sv and Cq rIAGs significantly elevated phosphorylation specific to their species, and in a dose-dependent manner. Mr rIAG was tested on Macrobrachium australiense (Ma), eliciting a similar response. Moreover, using bioinformatics analyses of the de novo assembled spiny lobster transcriptome, we identified a spiny lobster tyrosine kinase insulin receptor (Sv-TKIR). We validated this discovery with a receptor activation assay in COS-7 cells expressing Sv-TKIR, using a reporter SRE-LUC system designed for RTKs, with each of the rIAG proteins acting as the activation ligand. Using recombinant proteins, we aim to develop specific tools to control sexual development through the administration of IAG within the critical sexual differentiation time window. The biologically active rIAGs generated might facilitate commercially feasible solutions for the long sought techniques for sex-change induction and monosex population culture in crustaceans and shed new light on the physiological mode of action of IAG in crustaceans.

Against a backdrop of food insecurity, the farming of decapod crustaceans is a rapidly expanding and globally significant source of food protein. Sagmariasus verreauxi spiny lobster, the subject of this study, are decapods of underdeveloped aquaculture potential. Crustacean neuropeptide G-protein coupled receptors (GPCRs) mediate endocrine pathways that are integral to animal fecundity, growth and survival. The potential use of novel biotechnologies to enhance GPCR-mediated physiology may assist in improving the health and productivity of farmed decapod populations. This study catalogues the GPCRs expressed in the early developmental stages, as well as adult tissues, with a view to illuminating key neuropeptide receptors. De novo assembled contiguous sequences generated from transcriptomic reads of metamorphic and post metamorphic S. verreauxi were filtered for seven transmembrane domains, and used as a reference for iterative re-mapping. Subsequent putative GPCR open reading frames (ORFs) were BLAST annotated, categorised, and compared to published orthologues based on phylogenetic analysis. A total of 85 GPCRs were digitally predicted, that represented each of the four arthropod subfamilies. They generally displayed low-level and non-differential metamorphic expression with few exceptions that we examined using RT-PCR and qPCR. Two putative CHH-like neuropeptide receptors were annotated. Three dimensional structural modelling suggests that these receptors exhibit a conserved extracellular ligand binding pocket, providing support to the notion that these receptors co-evolved with their ligands across Decapoda. This perhaps narrows the search for means to increase productivity of farmed decapod populations.

The Caribbean spiny lobster, Panulirus argus (Latreille, 1804) is a highly commercial species and comprises the largest spiny lobster fishery in the world. Although populations have declined throughout its range, there is little known regarding its diseases and pathogens. The objectives of this study were to provide illustrated and standardized methods for postmortem examination, and to describe baseline gross and microscopic pathology for P. argus. From July 2017–March 2019, a postmortem examination including comprehensive histological assessment was performed on 313 fishery-caught lobsters. Epibionts and lesions observed include branchial cirriped infestation (69%), branchial encysted nemertean worm larvae (23%), tail fan necrosis (11%), skeletal muscle necrosis (7%), antennal gland calculi (6%), branchial infarction (2%), and microsporidiosis (0.6%). This report confirms the rare prevalence of microsporidiosis in P. argus and describes nemertean worm larvae in the gill. This study also reports a condition resembling excretory calcinosis in spiny lobster. The methods and data produced by this study facilitate disease diagnosis and sustainable stock management of P. argus.

Spiny lobster post-larvae undertake an extensive migration from the open ocean to the coast, during which time their swimming is fueled solely by energy reserves accumulated through their preceding larval phase. We assessed the influence of future ocean temperatures on the swimming behavior and energy use of migrating post-larvae of Sagmariasus verreauxi, by experimentally swimming post-larvae for up to 6 days at three temperatures and measuring the lipid and protein used, and observing their time spent actively swimming. Increasing the temperature from 17 °C to 23 °C doubled the energy utilized by post-larvae while swimming, while also reducing the time they spent swimming by three times. Therefore, increasing ocean temperatures appear to greatly affect the energetic cost and efficiency of shoreward migration of post-larvae in this lobster species, with the potential to markedly impact post-larval recruitment into coastal populations under future scenarios of ocean warming.

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.

Observations of the cuticle of the Boston Spiny Lobster using scanning electron microscope (SEM) show that it is a natural biocomposite consisting of chitin fibers and sclerotic-protein matrix with hierarchical and helicoidal structure. The SEM images also indicate that there is a hole–pin joining structure in the cuticle. In this joining structure, the chitin fibers in the neighborhood of the joining holes continuously round the holes to form a fiber-round-hole distribution. The maximum pullout force of the fibers in the fiber-round-hole distribution, which is closely related to the fracture toughness of the cuticle, is investigated and compared with that of the fibers in non-fiber-round-hole distribution based on their representative models. It is revealed that the maximum pullout force of the fibers in the fiber-round-hole distribution is significantly larger than that of the fibers in the non-fiber-round-hole distribution, and that a larger diameter of the hole results in a larger difference in the maximum pullout forces of the fibers between the two kinds of the fiber distributions. Inspired by the fiber-round-hole distribution found in the cuticle, composite specimens with the fiber-round-hole distribution were fabricated with a special mold and process to mirror the fiber-round-hole distribution. The fracture toughness of the biomimetic composite specimens is tested and compared with that of the conventional composite specimens with the non-fiber-round-hole distribution. It is demonstrated that the fracture toughness of the biomimetic composite specimens with the fiber-round-hole distribution is significantly larger than that of the conventional composite specimens with the non-fiber-round-hole distribution.

Bacteriocins are ribosomally synthesized antimicrobial peptides, which have been found in diverse bacterial species of terrestrial origins and some from the sea. New bacteriocins with new characteristics, new origins and new applications are likely still awaiting discovery. The present study screened bacteria isolated from marine animals of interest to the aquaculture industry for antimicrobial and bacteriocin-like activities in order to uncover biodiversity of bacteriocin producers, and explore the potential application in aquaculture. In total, 24 of 100 screened isolates showed antimicrobial activities and 7 of these exerted bacteriocin-like activities. Sequencing of 16S rRNA genes identified the isolates as members of the six genera Proteus, Providencia, Klebsiella, Alcaligenes, Bacillus and Enterococcus. In some cases, further analysis of housekeeping genes, rpoB for Proteus and recA for Klebsiella, as well as biochemical tests was necessary for identification to species level, and some of the Proteus isolates may represent novel species. The seven bacteriocinogenic isolates showed a wide antimicrobial spectrum against foodborne and animal pathogens, which opens the way to their potential use as marine drugs and probiotics in food, aquaculture, livestock and clinical settings. As a case study, the protective effect of shortlisted bacteriocinogenic isolates were tested in aquaculture-raised spiny lobster (Panulirus ornatus) juveniles. A single-strain (Bacillus pumilus B3.10.2B) and a three-strain (B. pumilus B3.10.2B, Bacillus cereus D9, Lactobacillus plantarum T13) probiotic preparation were added to the feed of Panulirus ornatus juveniles, which were subsequently challenged with the pathogen Vibrio owensii DY05. Juveniles in the probiotic treatments displayed increased growth and reduced feed conversion rates after 60 days, and increased survival rate after pathogen challenge relative to the control. This study represents the first evidence of bacteriocin production by bacteria associated with lobster, tiger shrimp, snubnose pompano and cobia and the first description of V. owensii as a pathogen in P. ornatus juveniles.

Dietary supplementation with mannan oligosaccharide (MOS) has been demonstrated to provide significant benefits to growth and health of many species of fish and crustaceans. An investigation was made to assess the effect of MOS supplementation on the diet of the tropical lobster Panulirus homarus. The aim of the study was to test the effect of various dietary inclusion levels of MOS on growth, survival rate, intestinal morphology and bacterial abundance, haemocyte make-up, and tissue composition of P. homarus lobsters. A manufactured, semi-moist diet was used as a basal diet and supplemented with 0.2%, 0.4%, 0.6% and 0.8% MOS inclusion, and fed to lobsters with an initial mean weight of 0.16 g for 56 days. A typical fishery bycatch mixture was used as a reference control diet. Growth rate of lobsters was significantly higher for those fed diets supplemented with 0.4 and 0.6% MOS than those fed an un-supplemented diet or bycatch. Survival rates of lobsters ranged from 68 to 78%. The survival rate was significantly higher in lobsters fed diets with 0.2% and 0.4% MOS supplementation and bycatch, compared with those fed the basal or 0.6% MOS diet. There was no significant difference in differential haemocyte count of lobsters fed with various MOS levels or bycatch. The mid-gut surface area of lobsters fed diets supplemented with 0.2%, 0.4% and 0.6% MOS was significantly higher than that of lobsters fed on control, 0.8% MOS or bycatch. The total Vibrio count in the mid-gut of lobsters fed any MOS diet was significantly lower than that of lobsters fed either the control or the bycatch diet. There was no significant difference in protein, lipid or ash content in lobsters fed the different diets. There were significant correlations between concentrations of MOS in diets and lobster growth rates. Optimal MOS concentration was estimated to be 0.69% at day 14 (R2 = 76.79%), 0.55% at day 28 (R2 = 88.18%), 0.48% at day 42 (R2 = 97.72%) and 0.47% at day 56 (R2 = 94.42%), demonstrating a decreasing requirement for MOS with increasing lobster size. In conclusion, mannan oligosaccharide supplementation appears to be effective for improving growth, survival and gastro-intestinal health of the lobster, P. homarus.

The ability of lobsters to recover from stress is an important factor influencing growth and survival in aquaculture; however, there is limited information on the metabolic responses of spiny lobsters to stress. This study determined oxygen consumption rate ( M ˙ O 2 ) and ammonia-N excretion of juvenile spiny lobsters Sagmariasus verreauxi in response to handling, anaesthesia, anaesthesia then force feeding, and exercising to exhaustion in order to determine the recovery time and amount of oxygen required to recover from common sources of stress in aquaculture. Lobsters required 0.29 ± 0.24 mg O2 g DM− 1 over a period of 2.5 ± 1.40 h to recover from handling. Anaesthesia with 2-phenoxyethanol immobilised lobsters but they required 0.72 ± 0.36 mg O2 g DM− 1 over a period of 4.5 ± 1.77 h to recover, indicating that 2-phenoxyethanol causes substantial stress. Force feeding lobsters anaesthetised with 2-phenoxyethanol or clove oil did not result in a higher increase of M ˙ O 2 compared to 2-phenoxyethanol anaesthesia alone. Recovery from exercise to exhaustion required 1.84 mg O2 g DM− 1 over a period of 10.17 ± 0.70 h. Ammonia-N excretion, measured 24 h post-treatment, and atomic O:N ratios were not significantly different between treatments and indicated lobsters in all groups had reached the same level of recovery 24 h post-treatment. Overall, the study demonstrated that lobsters recover rapidly from handling. Conversely, the increase in M ˙ O 2 induced by 2-phenoxyethanol demonstrated this is not an effective anaesthetic for reducing recovery periods and physiological stress associated with handling. However, 2-phenoxyethanol and clove oil eliminated further stress associated with force feeding, indicative of an invasive research procedure. Exercise to exhaustion resulted in the largest magnitude increase in M ˙ O 2 and longest recovery period. It is therefore recommended that aquaculture handling procedures take precautions to limit activity and are carried out without anaesthesia to facilitate more rapid recovery of lobsters.

Intraspecific analyses of the relationship between metabolic rate and mass have rarely been considered during complete ontogeny. Spiny lobsters are fascinating candidates to examine metabolic changes during ontogeny because their life cycle includes an extended planktonic, nektonic, and benthic life stages. The effect of body mass on metabolic rates, aerobic scope, and ammonia-N excretion of Sagmariasus verreauxi juveniles were examined to determine energetic demands through juvenile development. Mass-independent routine oxygen consumption increased allometrically during juvenile development with a mass scaling exponent of 0.83. The mass scaling exponent of active metabolism (0.81) was reduced compared to standard metabolism (0.91) of juvenile lobsters. The aerobic scope of juvenile lobsters decreased with larger body mass. To examine if the mass scaling exponent varies with ontogeny, we compared our data with previous measurements made with larvae of the same species. Comparison between mass scaling exponents showed they were higher for phyllosoma (0.97) compared to juvenile (0.83) development. Higher scaling exponents for phyllosoma may be attributed to increased growth rates of phyllosoma compared to juveniles, which increase oxygen consumption due to the higher energy cost of growth. The mass scaling exponent for complete ontogeny (0.91) of S. verreauxi was larger than the commonly cited 0.67 (1/3) and 0.75 (3/4) mass scaling exponents, indicating that species-specific differences can be a large factor affecting allometric relationships of animals.

Mass propagation of spiny lobster phyllosoma is a critical bottleneck for the development of spiny lobster aquaculture. Eastern spiny lobster Sagmariasus verreauxi has been identified as a good candidate for temperate culture with a relatively short life cycle lasting 6 to 8 months and 17 stages. We investigated the development and functionality of the filter-press of S. verreauxi phyllosoma to better understand the size and quantity of dietary particles ingested over the entire larval cycle. Filter-press morphometrics were obtained from live phyllosoma larvae throughout development, and filtration capacity was ascertained using fluorescent microspheres and the rare earth metal markers yttrium oxide and ytterbium oxide. The filter-press of phyllosoma larvae was fully functional from stage 3, fully developed from stage 4, and the numbers (8 to 50), lengths (79 to 384 μm), and widths (14.8 to 20.3 μm) of food grooves increased with the development stage to accommodate a larger volume of ingested food. In contrast, there was no change in the widths of the lower ampullary setae (1.28 ± 0.16 μm) or the widths of the gap between these setae (0.91 ± 0.07 μm). Lower ampullary setae were arranged in a single layer perpendicular to the food grooves and restricted the entry of food particles into the digestive gland. From stage 3, the filter-press selectively excluded more than 99% of particles of > 1 μm from reaching the digestive gland and demonstrated that only particles of ≤ 1 μm are available for digestion. In the absence of a gastric mill, phyllosoma larvae have no obvious structure for internally grinding prey and the present study shows that phyllosoma larvae are well adapted to feed from the body fluids of wild preys over their entire larval cycle. The results have important practical applications for the development of formulated feeds for spiny lobster larvae.

Operant discrimination learning has been extensively utilized in the study on the perceptual ability of animals and their higher order brain functions. We tested in this study whether American lobster Homarus americanus, which was previously found to possess ability of operant learning with claw gripping, could be trained to discriminate light stimuli of different intensities. For the current purpose, we newly developed a PC-controlled operant chamber that allowed the animal under a body-fixed condition to perform operant reward learning with claw gripping. Lobsters were first reinforced when they gripped the sensor bar upon presentation of a light cue. Then they were trained to grip the bar only when the light stimulus of a specific intensity was presented to obtain food reward while the stimuli of three different intensities including the reinforced one were presented in a random order. Finally, they were re-trained to grip the bar only when the light stimulus of another intensity that was not rewarded in the preceding training to obtain food while other intensities including the one that was rewarded previously were not rewarded any more. In these training procedures, the operant behavior occurred more frequently in response to the rewarded cue than to the non-rewarded one. The action latency for the reinforced stimuli showed a significant decrease in the course of training. These data demonstrate that lobsters can be trained with the light cues of different intensity as discriminative stimuli under a restrained condition that would allow application of electrophysiological techniques to the behaving subjects.

The slipper lobsters belong to the family Scyllaridae which contains a total of 20 genera and 89 species distributed across four subfamilies (Arctidinae, Ibacinae, Scyllarinae, and Theninae). We have collected nucleotide sequence data from regions of five different genes (16S, 18S, COI, 28S, H3) to estimate phylogenetic relationships among 54 species from the Scyllaridae with a focus on the species rich subfamily Scyllarinae. We have included in our analyses at least one representative from all 20 genera in the Scyllaridae and 35 of the 52 species within the Scyllarinae. Our resulting phylogenetic estimate shows the subfamilies are monophyletic, except for Ibacinae, which has paraphyletic relationships among genera. Many of the genera within the Scyllarinae form non-monophyletic groups, while the genera from all other subfamilies form well supported clades. We discuss the implications of this history on the evolution of morphological characters and ecological transitions (nearshore vs. offshore) within the slipper lobsters. Finally, we identify, through ancestral state character reconstructions, key morphological features diagnostic of the major clades of diversity within the Scyllaridae and relate this character evolution to current taxonomy and classification.

Concentrations of polycyclic aromatic hydrocarbons were determined in species of commercial fish and lobsters following an oil-spill just off the protected Madagascan coastline. Samples were collected along the coastline within and outside the affected area. Summed PAH concentrations ranged from 1.9 μg kg(-1) to 63 μg kg(-1) wet weight, but with no higher molecular weight PAHs (202 Da) being detected. All concentrations of benzo[a]pyrene, benz[a]anthracene and dibenz[a,h]anthracene were0.1 μg kg(-1) wet weight, well within the EU and UK set limits for the protection of human health. Additionally, samples were calculated as the benzo[a]pyrene toxic equivalency quotient (TEQ) and found to be well below the level of concern in relation to health of human consumers. Evaluation of the biota PAH data indicated the origin of PAH was predominantly petrogenic with80% arising from oil sources. Profile studies indicate a low-level multisource petrogenic contamination probably representing a pre-spill background for the area.

Nitric oxide (NO) is a short-lived radical generated by nitric oxide synthases (NOS). NO is involved in a variety of functions in invertebrates, including host defense. In a previous study, we isolated and sequenced for the first time the NOS gene from hemocytes of Panulirus argus, demonstrating the inducibility of this enzyme by lipopolysaccharide (LPS) in vitro. In the present work, lobster hemocytes and gills exposed to Escherichia coli O55:B5 LPS showed an increase in both NOS activity and NOS gene expression in vivo. This response was dose and time dependent. The 3D NOS structure was predicted by comparative modeling showing the oxygenase and reductase domains. These domains contain the conserved binding motifs of NOS already found in a variety of organisms. The 3D structure prediction analysis allowed the selection of a fragment of 666 bp that was cloned and subsequently expressed in E. coli BL21, in which a recombinant product of around 31 KDa was obtained. Hyperimmune serum obtained from immunized rabbits was tested and employed to specifically detect the recombinant polypeptide or the endogenous NOS from lobster hemocytes by western blot and immunofluorescence. This study contributes to enlarge the existing knowledge related to NOS structure and NOS participation in the immune response in lobsters. The evaluation of an antibody capable to recognize NOS from lobsters constitutes a novel and interesting tool for the implementation of further studies on NOS functions in crustaceans.

Linuparus White, 1847 comprises three extant species, Linuparus trigonus (Von Siebold, 1824), L. sordidus Bruce, 1965 , and L. somniosus Berry and George, 1972 , as well as 32 fossil species. Most fossil records are from North America and Europe, but the extant species are all confined to the Indo-West Pacific. Different colour forms in L. trigonus and L. sordidus have been noted, with Northern Hemisphere specimens generally darker in colour for both species. The phylogenetic relationships of the extant Linuparus species, including the colour forms, were investigated using mitochondrial 12S rRNA and COI gene sequence analysis. We found no genetic evidence to differentiate the colour morphs of L. sordidus, but the two colour forms of L. trigonus were clearly distinct at the species level. This is supported morphologically by a consistent difference in the shape of the thoracic sternum between the two forms. The paler coloured Southern Hemisphere form is described as a new species, L. meridionalis. Phylogenetic analysis shows that L. trigonus and L. meridionalis sp. nov. are derived sister taxa, while L. somniosus is basal within the genus. Thus the present results support the previous hypothesis that Linuparus was originated in shallow water.

Cross-linking with a bifunctional reagent and subsequent SDS gel electrophoresis is a simple but effective method to study the symmetry and arrangement of subunits in oligomeric proteins. In this study, theoretical expressions for the description of cross-linking patterns were derived for protein homohexamers through extension of the method used for tetramers by Hajdu et al. (1976). The derived equations were used for the analysis of cross-linking by glutardialdehyde of four protein hexamers: beef liver glutamate dehydrogenase (GDH), jack bean urease, hemocyanin from the spiny lobster Panulirus pencillatus (PpHc), Escherichia coli glutamate decarboxylase (GDC) and for analysis of published data on the cross-linking of hexameric E. coli rho by dimethyl suberimidate. Best fit models showed that the subunits in the first four proteins are arranged according to D(3) symmetry in two layers, each subunit able to cross-link to three neighboring subunits for GDH and urease, or to four for PpHc and GDC. The findings indicate a dimer-of-trimers eclipsed arrangement of subunits for GDH and urease and a trimer-of-dimers staggered one for PpHc and GDC. In rho, the subunits are arranged according to D(3) symmetry in a trimer-of-dimers ring. The conclusions from cross-linking of GDH and GDC, PpHc and rho are consistent with results from X-ray crystal structure, those for urease with findings from electron microscopy.

The effects of dietary mannan oligosaccharide (MOS) (Bio-Mos, Alltech, USA) on the growth, survival, physiology, bacteria and morphology of the gut and immune response to bacterial infection of tropical rock lobsters (Panulirus ornatus) juvenile were investigated. Dietary inclusion level of MOS at 0.4% was tested against the control diet (trash fish) without MOS inclusion. At the end of 56 days of rearing period, a challenged test was also conducted to evaluate the bacterial infection resistant ability of the lobsters fed the two diets. Lobster juvenile fed MOS diet attained 2.86 +/- 0.07 g of total weigh and 66.67 +/- 4.76% survival rate which were higher (P < 0.05) than the lobsters fed control diet (2.35 +/- 0.14 g total weight and 54.76 +/- 2.38% survival rate, respectively) thus providing the higher (P < 0.05) specific growth rate (SGR) and average weekly gain (AWG) of lobsters fed MOS diet. Physiological condition indicators such as wet tail muscle index (Tw/B), wet hepatosomatic index (Hiw) and dry tail muscle index (Td/B) of the lobsters fed MOS supplemented diet were higher (P < 0.05) than that of the lobsters fed the control diet. Bacteria in the gut (both total aerobic and Vibrio spp.) and gut's absorption surface indicated by the internal perimeter/external perimeter ratio were also higher (P < 0.05) when the lobsters were fed MOS diet. Lobsters fed MOS diet were in better immune condition showed by higher THC and GC, and lower bacteraemia. Survival, THC, GC were not different among the lobsters fed either MOS or control diet after 3 days of bacterial infection while bacteraemia was lower in the lobsters fed MOS diet. After 7 days of bacterial infection the lobsters fed MOS diet showed higher survival, THC, GC and lower bacteraemia than the lobsters fed the control diet. The experimental trial demonstrated the ability of MOS to improve the growth performance, survival, physiological condition, gut health and immune responses of tropical spiny lobsters juveniles.

Carotenoprotein, containing astaxanthin as the prosthetic group was extracted from the carapace shells of the lobster, Jasus lalandii. The material, with a molecular mass of around 400,000 Da and lambda(max) 525 nm (alpha-crustacyanin), was readily converted under the conditions used for extraction and on standing to the subunit (beta-crustacyanin) of molecular mass around 40,000 Da and lambda(max) 560 nm. The latter complex was stable up to 65 degrees C and between pH 5 and 8. Both the large molecule and the subunit showed the reversible bathochromic shift (lambda(max) approximately 480 nm) after being subjected to conditions of heat above 65 degrees C and below 85 degrees C and pH between 3 and 10, which alter the natural protein orientation. Above 85 degrees C and outside of the pH range, the shift was irreversible. Crustacyanin from J. lalandii differed from the blue crustacyanin of the Homarus species with respect to visible spectra (lambda(max) 632 nm and lambda(max) 585 nm) and by displaying a hypsochromic shift between the alpha- and beta-crustacyanin. Stability of the alpha-crustacyanin and affinity to DE52 also differed. Yellow protein, with a lambda(max) 409/410 nm as described for the Homarus species was not detected in this J. lalandii study, although it may have been co-eluted. The properties of the extracted complex show potential for its use as a natural water soluble food colourant or temperature sensitive indicator.

Lipid, fatty acid and protein content were determined individually on 7 phyllosomata, 69 clear pueruli, 286 pre-moult pueruli, and 86 juvenile western rock lobster (WRL) collected from four locations between the settlement seasons 2000 to 2006 to evaluate compositional changes during the non-feeding puerulus stage. Only the lipid content, particularly the phospholipids, decreased significantly with development. Protein declined sharply following moult to the juvenile. PL comprised between 86-94% of total lipid in all animals, and declined most between phyllosomata and clear pueruli (238.5 to 121.4 mg g(-1) DW) (p

We compared two indices of nutritional condition, the relative weight of the digestive gland (RWDG) and the ratio of weight to carapace length (weight/CL ratio), of the first three benthic phases, algal juveniles (7–25 mm CL), postalgal juveniles (25–45 mm CL), and subadults (45–79 mm CL) of Caribbean spiny lobster ( Panulirus argus ) dwelling in a shelter-poor reef lagoon before (1995) and after (2005) deployment of large artificial shelters (“casitas”) in 1998. To provide an experimental baseline, in 2005 we also conducted a laboratory experiment to assess the effect of starvation on both nutritional indices. Through 25-day trials, starvation significantly decreased the mean RWDG of all three juvenile phases, but did not affect their weight/CL ratios. In the field, casitas significantly enhanced juvenile lobsters relative to control sites and to prior values, but between 1995 and 2005 there was a significant decrease in condition of algal juveniles (both indices) and postalgal juveniles (only the RWDG), but not of subadults. These results would appear to counter previous findings that, for P. argus , food is not limiting and nutritional condition is not density-dependent. However, in 2000, a new pathogenic virus that has a distinct predilection for small juveniles, Panulirus argus Virus 1 (PaV1), emerged in the reef lagoon. Among other effects, this lethal virus causes a marked atrophy of the digestive gland. Through the laboratory trials and in the field, prevalence of visibly diseased individuals was highest in algal juveniles and lowest in subadults. These individuals were excluded from statistical analyses, but the disease progression may affect the digestive gland progressively given that newly infected lobsters take several weeks to become visibly diseased. Up to 2006, PaV1 had not significantly impacted lobster density on casita sites, but because P. argus is highly gregarious and transmission of PaV1 appears to be mainly by contact, futher investigation is required. These issues are relevant to enhancement of spiny lobsters and assessment of ecological impacts of marine diseases.

Flash photolysis and K-edge x-ray absorption spectroscopy (XAS) were used to investigate the functional and structural effects of pH on the oxygen affinity of three homologous arthropod hemocyanins (Hcs). Flash photolysis measurements showed that the well-characterized pH dependence of oxygen affinity (Bohr effect) is attributable to changes in the oxygen binding rate constant, k(on), rather than changes in k(off). In parallel, coordination geometry of copper in Hc was evaluated as a function of pH by XAS. It was found that the geometry of copper in the oxygenated protein is unchanged at all pH values investigated, while significant changes were observed for the deoxygenated protein as a function of pH. The interpretation of these changes was based on previously described correlations between spectral lineshape and coordination geometry obtained for model compounds of known structure (Blackburn, N. J., Strange, R. W., Reedijk, J., Volbeda, A., Farooq, A., Karlin, K. D., and Zubieta, J. (1989) Inorg. Chem., 28, 1349-1357). A pH-dependent change in the geometry of cuprous copper in the active site of deoxyHc, from pseudotetrahedral toward trigonal was assigned from the observed intensity dependence of the 1s --4p(z) transition in x-ray absorption near edge structure (XANES) spectra. The structural alteration correlated well with increase in oxygen affinity at alkaline pH determined in flash photolysis experiments. These results suggest that the oxygen binding rate in deoxyHc depends on the coordination geometry of Cu(I) and suggest a structural origin for the Bohr effect in arthropod Hcs.

Changes in major digestive enzymes through developmental and molt stages were studied for the spiny lobster Panulirus argus. There were significant positive relationships between specific activity of trypsin and amylase enzymes and lobster size, whereas esterase and lipase specific activities decreased as lobsters aged. No relationship was found between amylase/trypsin ratio and lobster size. Positive trends were found, however, for trypsin/lipase and amylase/lipase ratios. Results suggest that changes in enzyme activity respond to the lobsters' physiological needs for particular dietary components although multivariate analysis suggested that enzyme activities could be not totally independent of diet. On the other hand, the pattern of changes of major enzyme activities through molt cycle was similar for most enzymes studied. Following molt, trypsin, chymotrypsin, amylase, and lipase activities gradually increased to maximal levels at late intermolt (C4) and premolt (D). There were no variations in the electrophoretic pattern of digestive enzymes through developmental and molt stages and thus, it is demonstrated that regulation is exerted quantitatively rather than qualitatively. Further studies on the effect of other intrinsic and extrinsic factors on digestive enzyme activities are needed to fully understand digestive abilities and regulation mechanisms in spiny lobsters.

Pathogens can profoundly impact the ecology of the organisms they infect through changes in host behavior that influence demographic processes. For example, juvenile Caribbean spiny lobsters (Panulirus argus Latreille) infected with the PaV1 virus (Panulirus argus Virus 1) are avoided by their normally social conspecifics, which alters local spatial distributions and presumably rates of disease transmission. PaV1 infections are nearly always lethal, but prior to succumbing to the disease, infection may impact other host dynamics (e.g., movement, growth, or survival) that effect transmission of the virus. We used mark-recapture surveys and laboratory studies to determine the impact of PaV1 infection on lobster movement and physiological condition, and in turn, the effect of condition on susceptibility to infection. Significantly more healthy lobsters were recaptured than heavily diseased lobsters in 5-d mark-recapture surveys, indicating either greater emigration or greater mortality of infected lobsters. Results of a laboratory bioassay suggest that lobsters with early-stage infections moved at an equivalent rate to healthy lobsters, however, as infection progressed, lobsters inoculated with PaV1 moved less than healthy lobsters ultimately remaining sedentary. Infected lobsters captured in nature also had significantly lower hemolymph (blood) serum protein values, indicating poor physiological condition. However, poor condition did not predispose lobsters to greater risk of infection because prevalence was not different between starved lobsters and healthy lobsters challenged with PaV1. Although lobsters infected with PaV1 eventually become sedentary and of poor nutritional condition, during the early stages of infection they remain active and are thus capable of dispersing the virus throughout the population. © 2008 Elsevier B.V. All rights reserved.

Hemocyanin and phenoloxidase belong to the type-3 copper protein family, sharing a similar active center whereas performing different roles. In this study, we demonstrated that purified hemocyanin (450 kDa) from the spiny lobster Panulirus argus shows phenoloxidase activity in vitro after treatment with trypsin, chymotrypsin and SDS (0.1% optimal concentration), but it is not activated by sodium perchlorate or isopropanol. The optimal pHs of the SDS-activated hemocyanin were 5.5 and 7.0. Hemocyanin from spiny lobster behaves as a catecholoxidase. Kinetic characterization using dopamine, L-DOPA and catechol shows that dopamine is the most specific substrate. Catechol and dopamine produced substrate inhibition above 16 and 2 mM respectively. Mechanism-based inhibition was also evidenced for the three substrates, being less significant for L-DOPA. SDS-activated phenoloxidase activity is produced by the hexameric hemocyanin. Zymographic analysis demonstrated that incubation of native hemocyanin with trypsin and chymotrypsin, produced bands of 170 and 190 kDa respectively, with intense phenoloxidase activity. Three polypeptide chains of 77, 80 and 89 kDa of hemocyanin monomers were identified by SDS-PAGE. Monomers did not show phenoloxidase activity induced by SDS or partial proteolysis.

We examined the histopathological and hematological response of the Caribbean spiny lobster to experimentally induced infections with Panulirus argus Virus 1 (PaV1). The fixed phagocytes in the hepatopancreas were the primary sites of PaV1 infection in spiny lobsters. Fixed phagocytes were activated in early infections. However, as the disease progressed, the fixed phagocytes became infected and eventually lysed. Infected cells were subsequently observed in the hepatopancreas, gill, heart, hindgut, glial cells around the ventral nerves, and in the cuticular epidermis and foregut. In advanced infections, spongy connective tissues were heavily infected, as were glial cells around the optic nerves. The structure of the hepatopancreas was significantly altered as the disease progressed. The hemal sinuses among the hepatopancreatic tubules filled with massive amounts of cellular aggregates, including infected circulating hemocytes and spongy connective tissues. Atrophy of the hepatopancreatic tubules occurred in the late stage of viral infection. The virus caused significant decreases in total hemocyte counts and significantly altered several constituents in the hemolymph lysates of diseased lobsters, including: glucose, phosphorus, and triglycerides.

Based on data series for 8 consecutive years of settlement of post-larvae of the California spiny lobster Panulirus interruptus , its relationship to commercial catch was examined. The purpose was to establish a database to build a model for prediction of commercial catches of this lobster. The study area is a rocky shelf southwest of the bay entrance of Bahia Tortugas in Baja California Sur. A set of artificial collectors was placed at this site to obtain postpuerulus and early juveniles settlement for each month. Different simple regression analyses were performed on annual series of mean rates of post-larval settlement vs. commercial catches by the Bahia Tortugas fishing cooperative. We tested several delay periods between the series (4–8 years to approximate the age of fishery recruitment) to find the best-fit relationship. The relationship with a 5-year delay showed the highest positive correlation (0.7925) and was statistically significant. This delay period was used to regress the data for the years 1993–2001. The results showed that variations in the abundance of settled post-larvae in a given year are reflected in the commercial catch 5 years later and exhibit the same upward or downward trend. The one exception was the high post-larval settlement rate in 1997–1998, coinciding with the El Nino event, which was not mirrored by a similar increase 5 years later in the 2002–2003 catch. The pre-recruit index seems a very promising tool for predicting catches for this lobster fishery.

The high export value of the Indian spiny lobster Panulirus homarus increasingly attracts the aquaculturists for farming and fattening. However, lack of knowledge on the effect of environmental parameters on the immune system of this animal could result in high mortality, which ultimately may cause major loss to the industry. Here, we report the effect of salinity (20, 25, 35, 40, and 45 per thousand), pH (5.0, 8.0, and 9.5), dissolved oxygen (DO) (1 and 5 mg L(-1)), and ammonia-N concentration (0, 0.5, 1.5 and 3 mg L(-1)) on the immune response of P. homarus measured in the haemolymph in terms of Total Haemocyte Count (THC), phenoloxidase (PO) activity, and NBT-reduction. Our data showed significant reduction (P0.05) in THC, and NBT-reduction at lower (20 per thousand) and higher (45 per thousand) salinities. However, PO activity showed significant disparity, showing an increasing trend from 20 to 45 per thousand. Significant reduction (P0.05) in THC and PO activity under acidic and alkaline conditions, under hypoxic condition (1 mg L(-1)), and at the higher ammonia-N concentrations than their respective optimal conditions were observed. Thus, suggesting that extreme environmental parameters can induce modifications in the immune system of the spiny lobster P. homarus, which may enhance their susceptibility to opportunistic pathogens. The humoral parameters such as THC, PO activity, and NBT-reduction can be used as potential stress indicators for healthy management of spiny lobsters.

Spiny lobsters are prevalent in tropical and temperate seas and support some of the largest commercial fisheries in the world. Studies on the metabolic rate of spiny lobsters contribute to the understanding of the relation of these widely distributed crustaceans with their environment, to the design of holding facilities and to the development of future aquaculture technologies. The current study attempts to describe the oxygen consumption of juvenile lobsters Panulirus argus in relation with body weight, temperature and feeding. Individual routine metabolism increased with body weight according to the regression line Log 10 M O 2 (mg O 2 /h) = 0.754Log 10 W (g) − 0.678 ( r 2 = 0.71) whereas weight-specific respiration rate decreased according to the regression line Log 10 V O 2 (mg O 2 /kg/h) = − 0.774Log 10 W (g) + 3.395 ( r 2 = 0.81). The metabolism of juvenile P. argus showed a linear increase with temperature and this relationship was best described by the regression line V O 2 (mg O 2 /kg/h) = 6.183 T (°C) − 110.21 ( r 2 = 0.72). All regressions were found to be statistically significant. From the evaluated factors, feeding had the highest effect on oxygen consumption. The oxygen consumption increased after ingestion with the maximum within the first 5 h. Results are discussed in relation with their relevance for maintaining P. argus under captive conditions.

On-growing of wild-caught juvenile spiny (Palinuridae) lobsters to market size is an emerging aquaculture industry in Asia, Oceania and Central America but most notably in Vietnam where a US$90 million per annum industry flourishes. The sustainability of the supply of wild juveniles is an ever present threat to industry expansion, which ultimately may be overcome through successful life cycle closure. However, another and perhaps more imminent impediment is the present reliance on fresh fishery by-catch as a source of food for lobster culture. The decreasing availability and increasing cost of the by-catch, together with the downstream environmental impacts caused by this type of feeding, has focused attention on the need for more sustainable and environmentally-friendly pelleted feeds. Efforts to develop palatable and high performance pelleted dry feeds for spiny lobster grow-out have not met with a lot of success until recently. This paper reviews the progress that has been made and identifies where further research effort is needed.