Does Differential Receptor Distribution Underlie Variable Responses to a Neuropeptide in the Lobster Cardiac System?

Central pattern generators produce rhythmic behaviors independently of sensory input; however, their outputs can be modulated by neuropeptides, thereby allowing for functional flexibility. We investigated the effects of C-type allatostatins (AST-C) on the cardiac ganglion (CG), which is the central pattern generator that controls the heart of the American lobster, Homarus americanus, to identify the biological mechanism underlying the significant variability in individual responses to AST-C. We proposed that the presence of multiple receptors, and thus differential receptor distribution, was at least partly responsible for this observed variability. Using transcriptome mining and PCR-based cloning, we identified four AST-C receptors (ASTCRs) in the CG; we then characterized their cellular localization, binding potential, and functional activation. Only two of the four receptors, ASTCR1 and ASTCR2, were fully functional GPCRs that targeted to the cell surface and were activated by AST-C peptides in our insect cell expression system. All four, however, were amplified from CG cDNAs. Following the confirmation of ASTCR expression, we used physiological and bioinformatic techniques to correlate receptor expression with cardiac responses to AST-C across individuals. Expression of ASTCR1 in the CG showed a negative correlation with increasing contraction amplitude in response to AST-C perfusion through the lobster heart, suggesting that the differential expression of ASTCRs within the CG is partly responsible for the specific physiological response to AST-C exhibited by a given individual lobster.

Short-term exposure to hydrogen peroxide induces mortality and alters exploratory behaviour of European lobster (Homarus gammarus).

Bath treatment chemotherapeutants, used to control sea lice infestations in the salmonid aquaculture industry, are released directly into the marine environment around fish farms and pose a serious risk to non-target species, particularly crustaceans. Hydrogen peroxide (H2O2) is the most frequently used bath treatment chemotherapeutant on Norwegian fish farms, however, limited information is available on its toxicity to European lobsters (Homarus gammarus), a commercially important species at risk of exposure due to its distribution overlapping with salmon farm locations. The aim of this study was to investigate the lethal effects of H2O2 on pelagic (stage I-IV) larvae/post-larvae and its sub-lethal effects on the benthic stage V H. gammarus. To assess the lethal effects of H2O2, we carried out a series of 1 h toxicity tests and assessed mortality after a 24 h post-exposure period. Exposure to H2O2 was toxic to all pelagic larval stages tested, with estimated median lethal concentrations (LC50) of 177, 404, 665 and 737 mg/L for stage I, II, III and IV, respectively. These concentrations represent approximately 10, 23, 40 and 43%, of the recommended H2O2 concentrations used for delousing salmon on Norwegian fish farms, respectively. To assess the sub-lethal effects of H2O2 on H. gammarus, stage V juveniles were exposed to H2O2 at concentrations of 85, 170 and 510 mg/L for 1 h and shelter-seeking behaviour and mobility endpoints were assessed. Numerous behavioural parameters including distance travelled to shelter, time to locate shelter and the number of shelter inspections, were negatively affected in lobsters exposed to H2O2 when assessed immediately after the exposure period. However, no differences between control and exposed lobsters were detected after a 24 h post-exposure period. Our results demonstrate that short term exposures to H2O2 are lethal to pelagic H. gammarus life stages and can negatively affect the shelter seeking behaviour of benthic life stages, though these behavioural changes may be short-lived.

Acute and long-term manganese exposure and subsequent accumulation in relation to idiopathic blindness in the American lobster, Homarus americanus.

Manganese (Mn) is a hypoxic reactive metal commonly found in marine sediments. Under hypoxic conditions the metal becomes fully reduced to Mn2+ and is biologically available to the benthic community for uptake. Mn is also a potent neurotoxin and it may play a role in the etiology of idiopathic blindness that has been observed in American lobsters. An acute exposure study was designed to expose American lobster, Homarus americanus, to 0, 20, 80, 150, and 300 mg L-1 (ppm) for 96 hs to explore disparities in Mn accumulation among several tissues: optic nerve, brain, hepatopancreas, muscle, hemolymph, gill, and exoskeleton. These concentrations were based on realistic pore-water concentrations (20 mg L-1), high sediment concentrations (80 mg L-1), and unrealistically high concentrations to determine lethality (150 and 300 mg L-1). A positive correlation between Mn accumulation and exposure concentration was observed in all tissues examined. In the internal tissues, manganese concentrations showed a high affinity towards brain, optic nerve, and hemolymph. In the exoskeleton and gills, Mn concentrations were also high, possibly because of internal uptake as well as external adsorption. Concentrations of Mn in tissues from the acute exposure study followed the accumulation pattern: hemolymph > gill > exoskeleton > optic nerve > brain > hepatopancreas = muscle. A long-term exposure study lasting seven weeks was designed to investigate the potential link between high Mn exposure and idiopathic blindness, a condition that affects an estimated 50 % of the adult American lobster population off Southern New England (SNE), USA. A comparison of these exposure studies showed evidence of time-dependent Mn accumulation in brain, muscle, exoskeleton, and gill tissue. Although the relationship between Mn exposure and blindness was not apparent, there was a modest trend in the development of blindness (Chi-square, p = 0.102) in animals exposed to a high concentration (150 mg L-1) of the metal. With no mortalities occurring in the acute study and only one mortality in the long-term study, it is highly unlikely that Mn is acutely toxic to American lobsters at environmentally relevant concentrations. Its potential role in idiopathic blindness remains to be determined.

Impact of a Novel Cryoprotectant Blend on the Sensory Quality of Frozen Lobster (Homarus americanus).

Frozen storage of lobster meat (Homarus americanus) can result in undesirable quality changes that decrease consumer acceptability of these products. Current seafood industry methods use cryoprotective agents that contain phosphates including sodium tripolyphosphates (STPP). However, recent evidence suggests that cryoprotective mixtures that combine different carbohydrates and STPP can have equal or even greater cryoprotective properties compared to using STPP alone. The objective of this study was to compare the overall consumer acceptability of lobster meat stored for 6 months in different blends of these cryoprotective solutions. One hundred and seven panelists were recruited to score the acceptability of the lobster samples using nine-point hedonic scales. A check-all-that-apply (CATA) question containing 27 literature-informed, sensory descriptors was also used to identify terms frequently used to describe lobster meat. Analysis of variance analysis, indicated a significant increase for overall liking (22.1%, P < 0.0001), liking of flavor (23.6%, P < 0.0001) and texture (15.6%, P = 0.000) scores for samples stored in a novel carbohydrate blend plus sodium chloride (NaCl) and STPP compared to the water control. Subsequent penalty analysis revealed that overall liking scores were most positively associated with the attributes tender, sweet, moist and soft. Moreover, the attributes with the highest positive mean impact were more frequently used to describe lobster samples stored in solutions containing NaCl and the novel carbohydrate blend, as well as NaCl and STPP (Lobster-3 and Lobster-5 samples, respectively). PRACTICAL APPLICATION: The positive impact on the sensory quality of this novel blend of cryoprotective compounds (carbohydrates and NaCl) is proof of concept that this mixture is comparable, if not better than preservatives currently used by the seafood industry. Given the necessary regulatory approval and industry acceptance, lobster processors may consider this novel blend as a suitable alternative to freeze lobster products for up to 6 months.

Effects of Sublethal Chlorpyrifos Exposure on Postlarval American Lobster (Homarus americanus).

The organophosphate pesticide chlorpyrifos has been introduced to the marine environment via adsorption to agricultural soil runoff or as spray drift. Chlorpyrifos affects the survival of some larval decapod crustaceans, but no data exist on the impacts to the American lobster, Homarus americanus. The purpose of the present study was to assess the levels at which chlorpyrifos affects the survival of postlarval H. americanus. Using acute saltwater exposures, the 24- and 48-h median lethal concentrations were established for stage IV H. americanus (1.56 and 1.33 µg/L, respectively). Movement, acetylcholinesterase activity, intermoult period, specific growth rate, and moult increment were measured during exposure to sublethal concentrations. Movement patterns were assessed to establish a 48-h median inhibition concentration for cessation of normal movement (0.66 µg/L). Acetylcholinesterase activity was found to be inhibited immediately post-exposure to 0.50, 0.57, and 0.82 µg/L chlorpyrifos but could be recovered within a period (9-15 d) in clean seawater. Sublethal growth effects of increased intermoult period, decreased specific growth rate, and decreased moult increment were observed during exposure to an environmentally relevant concentration (0.82 µg/L). The present study suggests that H. americanus stage IV larvae were marginally less sensitive to chlorpyrifos compared with other decapods and that acute lethality of H. americanus postlarvae is not likely to occur with chlorpyrifos concentrations previously reported from aquatic environments. Environ Toxicol Chem 2019;38:1294-1301. © 2019 SETAC.

Plastic pollution affects American lobsters, Homarus americanus.

This paper provides the first record of ingestion of plastic debris by American lobster, Homarus americanus. Plastics particles, identified as rubber pieces, were found in the stomachs of 3 from 17 individuals of lobsters kept in laboratory conditions. Debris had evidence of cuts, what suggest they were actively consumed.

The effects of emamectin benzoate or ivermectin spiked sediment on juvenile American lobsters (Homarus americanus).

This study examined the effects of a range of ½-log concentrations of emamectin benzoate (commercially applied as SLICE®) and ivermectin (commercially applied as IVOMEC®) on juvenile American lobster, Homarus americanus. Phase I of the research assessed acute (up to 4 days) and chronic (30-day) toxicity of sediment dosed with the active ingredients emamectin benzoate (EMB) formulated as SLICE® or ivermectin (IVM) formulated as IVOMEC® at various nominal concentrations (EMB: 15, 48, 150, 475 and 1500 ng g-1 wet sediment; IVM: 3, 9.5, 30, 95 and 300 ng g-1 wet sediment) on juvenile Atlantic lobster (stages IV). Phase II evaluated sublethal effects (e.g., growth, moulting success) of all lobster surviving past the 30 day exposure period, over an additional 41 days. Chemical analysis of EMB and IVM in sediment samples from the exposure tanks revealed a strong linear association (R2 values 0.99 and 0.98 for EMB and IVM, respectively) between nominal dose and measured concentration of compound. EMB exposure concentrations at very high levels (≥ 343.3 ng g-1) were acutely toxic to juvenile lobster such that 100% of lobsters had died after 13 days of exposure. The maximum cumulative mortality of lobsters exposed to the highest concentrations of EMB and IVM was 100% after 10 days and 25 days, respectively. The 10-day LC50 estimates (±â€¯95% CI) for EMB and IVM were 250.23 ±â€¯90.4 and 212.14 ±â€¯202.64 ng g-1, respectively. Using abnormal behaviour as an indicator, the 15-day EC50 estimates (±â€¯95% CI) for EMB and IVM were 96.19 ±â€¯51.42 and 15.82 ±â€¯6.93 ng g-1, respectively. The NOEC (no observed effect concentration) for abnormal behaviour was 0.0 ng g-1 for each product and the LOEC (lowest observed effect concentration) was 8.8 and > 3.0 ng g-1 for EMB and IVM, respectively. Observations on sublethal effects included delayed moulting to stage VI and reduced growth at higher exposure concentrations for both therapeutants. Using failure to moult to stage V or VI as an indicator, the 15-day EC50 estimates (±â€¯95% CI) for EMB and IVM were 32.72 ±â€¯18.26 and 14.00 ±â€¯12.43 ng g-1, respectively. The NOEC for failure to moult to stage V only was 343.3 and 14.7 ng g-1 for EMB and IVM, respectively. Whereas, the LOEC was 1066.7 and > 61.0 ng g-1 for EMB and IVM, respectively. The concentrations of EMB and IVM tested in the present study were acutely toxic to juvenile lobster exposed to the highest dosages (343.3 and 1066.7 ng EMB g-1 and 61.0 and 300.0 ng IVM g-1). There was significant evidence of chronic toxicity, longer exposure increased mortality with LT50 values decreasing with increasing test material concentration.

Exposure to teflubenzuron negatively impacts exploratory behavior, learning and activity of juvenile European lobster (Homarus gammarus).

Infestations with salmon lice, a parasitic copepod, is a major problem in the salmon farming industry. Teflubenzuron is an in-feed pharmaceutical applied to control lice outbreaks; the standard medication is 10 mg per kg fish per day for seven days. Surveys reveal that teflubenzuron accumulates and persists in the sediment around fish farms and causes deformities and mortality in juvenile European lobster (Homarus gammarus), a species commonly found in the vicinity of salmon farms in Norway. To date, there is no information on sub-lethal effects of teflubenzuron on, for example, behavior. We conducted an experiment to assess possible difference in the shelter seeking behavior of teflubenzuron-exposed (N = 19) vs. not exposed (N = 19) H. gammarus juveniles. The teflubenzuron-exposed juveniles had been given very low concentrations, 1.7 µg per pellet twice per week for 113 days prior to this experiment. The concentration of teflubenzuron was estimated to be less than 1 ng/g lobster when they were tested in the behavior experiment. Animals were placed in a lane with a shelter at one end. Once a lobster had found and entered the shelter, they were repeatedly displaced back to the opposite end of the lane, for a total of 3 repeated runs per animal. Three of the exposed juveniles failed to settle in the shelter, and the remaining teflubenzuron-exposed animals took significantly more time to explore the environment and to find and recognize shelter. Furthermore, exposed lobsters also exhibited slower walking speed compared to the controls. These results demonstrate that teflubenzuron significantly reduces exploratory behavior, learning and activity of juvenile H. gammarus. Thus, exposure to teflubenzuron could increase predation mortality of juvenile lobsters in the wild.

Homarus gammarus (Crustacea: Decapoda) larvae under an ocean acidification scenario: responses across different levels of biological organization.

The present study evaluated the effects of exposure to different target pCO2 levels: control (C: 370µatm, pH=8.15) and ocean acidification (OA: 710µatm, pH=7.85) on development and biochemical responses related with oxidative stress and energy metabolism during the crustacean Homarus gammarus (L.) larval development, integrating different levels of biological organization. After hatching in the laboratory, larvae from the same female brood were exposed to the described conditions from hatching until reaching Stage III (last larval stage - 11days). H. gammarus larvae demonstrated some susceptibility when addressing the predicted pCO2 levels for 2100. Further analysis at the biochemical and physiological level highlighted the occurrence of oxidative stress in the OA scenario (Superoxide Dismutase reduction and higher DNA damage) that was followed by developmental effects, increased inter-moult period from SII to SIII and reduced growth. The extended exposure to these conditions may affect organisms' key life-cycle functions such as physiological resistance, growth, sexual maturation, or reproduction with implications in their future fitness and population dynamics.

Long-term microplastic retention causes reduced body condition in the langoustine, Nephrops norvegicus.

Microplastic represents a rising proportion of marine litter and is widely distributed throughout a range of marine habitats. Correspondingly, the number of reports of species containing microplastics increases annually. Nephrops norvegicus in the Firth of Clyde have previously been shown to retain large aggregations of microplastic fibres. The potential for N. norvegicus to retain plastic over an extended time period increases the likelihood of any associated negative impacts to the individual. This study represents the longest observation of the impacts of microplastic retention in invertebrates. We exposed N. norvegicus to plastic over eight months to determine the impacts of extended exposure. Over this period we compared the feeding rate, body mass, and nutritional state of plastic-fed N. norvegicus to that of fed and starved control groups. Following the experimental period, the plastic-fed langoustine contained microplastic aggregations comparable to those of small individuals from the Clyde Sea Area. Comparisons between fed, unfed and plastic-fed individuals indicated a reduction in feeding rate, body mass, and metabolic rate as well as catabolism of stored lipids in plastic contaminated animals. We conclude that N. norvegicus exposed to high levels of environmental microplastic pollution may experience reduced nutrient availability. This can result in reduced population stability and may affect the viability of local fisheries.

Forces generated during stretch in the heart of the lobster Homarus americanus are anisotropic and are altered by neuromodulators.

Mechanical and neurophysiological anisotropies mediate three-dimensional responses of the heart of ITALIC! Homarus americanus Although hearts ITALIC! in vivoare loaded multi-axially by pressure, studies of invertebrate cardiac function typically use uniaxial tests. To generate whole-heart length-tension curves, stretch pyramids at constant lengthening and shortening rates were imposed uniaxially and biaxially along longitudinal and transverse axes of the beating whole heart. To determine whether neuropeptides that are known to modulate cardiac activity in ITALIC! H. americanusaffect the active or passive components of these length-tension curves, we also performed these tests in the presence of SGRNFLRFamide (SGRN) and GYSNRNYLRFamide (GYS). In uniaxial and biaxial tests, both passive and active forces increased with stretch along both measurement axes. The increase in passive forces was anisotropic, with greater increases along the longitudinal axis. Passive forces showed hysteresis and active forces were higher during lengthening than shortening phases of the stretch pyramid. Active forces at a given length were increased by both neuropeptides. To exert these effects, neuropeptides might have acted indirectly on the muscle via their effects on the cardiac ganglion, directly on the neuromuscular junction, or directly on the muscles. Because increases in response to stretch were also seen in stimulated motor nerve-muscle preparations, at least some of the effects of the peptides are likely peripheral. Taken together, these findings suggest that flexibility in rhythmic cardiac contractions results from the amplified effects of neuropeptides interacting with the length-tension characteristics of the heart.

Transcriptional responses to teflubenzuron exposure in European lobster (Homarus gammarus).

Increasing use of pharmaceutical drugs to delouse farmed salmon raises environmental concerns. This study describes an experiment carried out to elucidate the molecular mechanisms of the antiparasitic drug teflubenzuron on a non-target species, the European lobster. Juvenile lobsters (10.3±0.9 mm carapace length) were fed two environmentally relevant doses of teflubenzuron, corresponding to 5 and 20% of a standard salmon medication (10 mg/kg day), termed low and high dose in this study. After 114 days of dietary exposure, whole-animal accumulation of teflubenzuron was determined. One claw from each animal was collected for transcriptional analysis. Overall, exposed animals showed low cumulative mortality. Six animals, two from the low dose treatment and four from the high dose, showed exoskeletal abnormalities (claw deformities or stiff walking legs). Residual levels of teflubenzuron in juvenile lobster were 2.7-fold higher in the high dose (282 ng/g) compared to the low dose treatment (103 ng/g). The transcriptional examination showed significant effects of teflubenzuron on 21 out of 39 studied genes. At the transcriptional level, environmentally relevant levels of the anti-salmon lice drug impacted genes linked to drug detoxification (cyp3a, cyp6a2, cyp302a, sult1b1, abcc4), cellular stress (hsp70, hsp90, chh), oxidative stress (cat, gpx3) and DNA damage (p53), as well as molting and exoskeleton regulation (chi3l1, ecr, jhl1, chs1, ctbs, gap65, jhel-ces1) in claw tissue (muscle and exoskeleton). In conclusion, teflubenzuron at sub-lethal levels can affect many molecular mechanisms in European lobster claws.

Distinct or shared actions of peptide family isoforms: II. Multiple pyrokinins exert similar effects in the lobster stomatogastric nervous system.

Many neuropeptides are members of peptide families, with multiple structurally similar isoforms frequently found even within a single species. This raises the question of whether the individual peptides serve common or distinct functions. In the accompanying paper, we found high isoform specificity in the responses of the lobster (Homarus americanus) cardiac neuromuscular system to members of the pyrokinin peptide family: only one of five crustacean isoforms showed any bioactivity in the cardiac system. Because previous studies in other species had found little isoform specificity in pyrokinin actions, we examined the effects of the same five crustacean pyrokinins on the lobster stomatogastric nervous system (STNS). In contrast to our findings in the cardiac system, the effects of the five pyrokinin isoforms on the STNS were indistinguishable: they all activated or enhanced the gastric mill motor pattern, but did not alter the pyloric pattern. These results, in combination with those from the cardiac ganglion, suggest that members of a peptide family in the same species can be both isoform specific and highly promiscuous in their modulatory capacity. The mechanisms that underlie these differences in specificity have not yet been elucidated; one possible explanation, which has yet to be tested, is the presence and differential distribution of multiple receptors for members of this peptide family.

Bacteriostatic suppression in Norway lobster (Nephrops norvegicus) exposed to manganese or hypoxia under pressure of ocean acidification.

Future ocean acidification (OA) and warming following climate change elicit pervasive stressors to the inhabitants of the sea. Previous experimental exposure to OA for 16 weeks at pH levels predicted for 2100 has shown to result in serious immune suppression of the Norway lobster, Nephrops norvegicus. The lobsters are currently affected by stressors such as periodical hypoxia inducing high levels of bioavailable manganese (Mn). Here, we aimed to investigate possible effects of interactions between OA and these stressors on total hemocyte counts (THCs) and on recovery of inoculated bacteria in the lobsters, measured as a proxy for bacteriostatic response. The effects were judged by following numbers of culturable Vibrio parahaemolyticus in hepatopancreas, 4 and 24h post inoculation in lobsters kept in replicate tanks with six different treatments: either ambient (pCO2∼500 µatm/pH∼8.1 U) or CO2-manipulated seawater (OA; pCO2∼1550 µatm/pH∼7.6 U) for 8 weeks. During the last 2 weeks, additional stress of either hypoxia (∼23% oxygen saturation) or Mn (∼9 mg L(-1)) was added except in control treatments. Our results showed clear effect on bacteriostatic response in Norway lobsters exposed to these stressors. In lobsters kept in ambient seawater without additional stressors, the number of culturable bacteria in hepatopancreas was reduced by ∼34%. In combined treatment of ambient seawater and hypoxia, the reduction was ∼23%, while in the Mn-exposed animals, there was no reduction at all. This was also the case in all OA treatments where mean numbers of culturable V. parahaemolyticus tended to increase. In lobsters from ambient seawater with or without hypoxia, the THC was not significantly different as was also the case in OA without additional stressors. However, in OA treatments combined with either hypoxia or Mn, THC was reduced by ∼35%. While the reduction of culturable V. parahaemolyticus in lobsters was clearly affected by these stressors, we found no notable effects on growth, survival or hemolytic properties of the bacteria itself. Thus, we conclude that this predicted stress scenario is beneficial for the pathogen in its interaction with the host. As OA proceeds, it may force the health of the ecologically and economically important N. norvegicus to a tipping point if exposed to more short-term stressors such as the periodical events of hypoxia and Mn. This could impact lobster condition and biomass and may as well increase the risk for bacterial transmission to consumers.

Sublethal exposure to azamethiphos causes neurotoxicity, altered energy allocation and high mortality during simulated live transport in American lobster.

In the Bay of Fundy, New Brunswick, sea lice outbreaks in caged salmon are treated with pesticides including Salmosan(®), applied as bath treatments and then released into the surrounding seawater. The effect of chronic exposure to low concentrations of this pesticide on neighboring lobster populations is a concern. Adult male lobsters were exposed to 61 ngL(-1) of azamethiphos (a.i. in Salmosan(®) formulation) continuously for 10 days. In addition to the direct effects of pesticide exposure, effects on the ability to cope with shipping conditions and the persistence of the effects after a 24h depuration period in clean seawater were assessed. Indicators of stress and hypoxia (serum total proteins, hemocyanin and lactate), oxidative damage (protein carbonyls in gills and serum) and altered energy allocation (hepatosomatic and gonadosomatic indices, hepatopancreas lipids) were assessed in addition to neurotoxicity (chlolinesterase activity in muscle). Directly after exposure, azamethiphos-treated lobsters had inhibition of muscle cholinesterase, reduced gonadosomatic index and enhanced hepatosomatic index and hepatopancreas lipid content. All these responses persisted after 24-h depuration, increasing the risk of cumulative impacts with further exposure to chemical or non-chemical stressors. In both control and treated lobsters exposed to simulated shipment conditions, concentrations of protein and lactate in serum, and protein carbonyls in gills increased. However, mortality rate was higher in azamethiphos-treated lobsters (33 ± 14%) than in controls (2.6 ± 4%). Shipment and azamethiphos had cumulative impacts on serum proteins. Both direct effects on neurological function and energy allocation and indirect effect on ability to cope with shipping stress could have significant impacts on lobster population and/or fisheries.

Mortality and deformities in European lobster (Homarus gammarus) juveniles exposed to the anti-parasitic drug teflubenzuron.

This study describes experiments carried out to examine effects of the antiparasitic drug teflubenzuron, used in delousing farmed salmon, on a non-target species, the European lobster (Homarus gammarus). Juvenile lobsters were fed two doses of teflubenzuron, 10 and 20mg/kg successively for 7 days corresponding to a standard medication of the fish (10mg/kg day) and twice the standard dose (20mg/kg day). Monitoring lasted 3 months to include at least one moulting period for all individuals. Cumulative mortality was higher in all replicates given medicated feed compared with the control group. Mean cumulative mortality for each dosing was 41 ± 13% for 10mg/kg and 38 ± 8% for 20mg/kg, i.e. no difference. Drug residue was analysed in all juveniles that died, in addition to 12 juveniles at day 8 and the first 12 surviving lobsters. A decline in concentration of teflubenzuron from over 8,000 ng/g (day 5) to 14 ng/g (day 70) was observed in the juveniles that died during the experiment. Twelve individuals that died contained 82 ng/g or less whereas the mean concentration in the first 12 lobsters that survived moulting was 152 ng/g. Following a single oral administration, the half-life of teflubenzuron in lobster was estimated to 3.4 days and the initial concentration (C0) to 515 ng/g at time t0. At the end of the study a considerable number of juvenile lobsters were observed with deformities in various organs; carapace, walking legs, cheliped, tail fan, abdomen and antenna. The occurrence of observed deformities varied from 0 to 15% in treated replicates and will most likely affect ability to locate and consume food (antenna, claw and walking legs), respiration (carapace) and ability to move/swim (walking legs, tail fan and abdomen). In total, the mortality and senescent damages were close to 50% in all replicates. Juveniles that survived medication without deformities however, moulted and increased in size at each moult equally well as the unmedicated controls.

Coordination of distinct but interacting rhythmic motor programs by a modulatory projection neuron using different co-transmitters in different ganglia.

While many neurons are known to contain multiple neurotransmitters, the specific roles played by each co-transmitter within a neuron are often poorly understood. Here, we investigated the roles of the co-transmitters of the pyloric suppressor (PS) neurons, which are located in the stomatogastric nervous system (STNS) of the lobster Homarus americanus. The PS neurons are known to contain histamine; using RT-PCR, we identified a second co-transmitter as the FMRFamide-like peptide crustacean myosuppressin (Crust-MS). The modulatory effects of Crust-MS application on the gastric mill and pyloric patterns, generated in the stomatogastric ganglion (STG), closely resembled those recorded following extracellular PS neuron stimulation. To determine whether histamine plays a role in mediating the effects of the PS neurons in the STG, we bath-applied histamine receptor antagonists to the ganglion. In the presence of the antagonists, the histamine response was blocked, but Crust-MS application and PS stimulation continued to modulate the gastric and pyloric patterns, suggesting that PS effects in the STG are mediated largely by Crust-MS. PS neuron stimulation also excited the oesophageal rhythm, produced in the commissural ganglia (CoGs) of the STNS. Application of histamine, but not Crust-MS, to the CoGs mimicked this effect. Histamine receptor antagonists blocked the ability of both histamine and PS stimulation to excite the oesophageal rhythm, providing strong evidence that the PS neurons use histamine in the CoGs to exert their effects. Overall, our data suggest that the PS neurons differentially utilize their co-transmitters in spatially distinct locations to coordinate the activity of three independent networks.

Disease effects on lobster fisheries, ecology, and culture: overview of DAO Special 6.

Lobsters are prized by commercial and recreational fishermen worldwide, and their populations are therefore buffeted by fishery practices. But lobsters also remain integral members of their benthic communities where predator-prey relationships, competitive interactions, and host-pathogen dynamics push and pull at their population dynamics. Although lobsters have few reported pathogens and parasites relative to other decapod crustaceans, the rise of diseases with consequences for lobster fisheries and aquaculture has spotlighted the importance of disease for lobster biology, population dynamics and ecology. Researchers, managers, and fishers thus increasingly recognize the need to understand lobster pathogens and parasites so they can be managed proactively and their impacts minimized where possible. At the 2011 International Conference and Workshop on Lobster Biology and Management a special session on lobster diseases was convened and this special issue of Diseases of Aquatic Organisms highlights those proceedings with a suite of articles focused on diseases discussed during that session.

Histological intersex (ovotestis) in the European lobster Homarus gammarus and a commentary on its potential mechanistic basis.

This paper provides the first report of the intersex (ovotestis) condition in the European lobster Homarus gammarus. A single specimen (10% of males sampled) presenting the condition was discovered as part of routine sampling, from the Weymouth Bay region of the English Channel, UK. The lobster presented externally as a male, but upon histological examination was seen to contain an ovotestis, containing elements of both male and female gonadal tissue. Previtellogenic oocytes were present in several otherwise normal seminiferous tubules throughout the testis. The seminiferous tubules were also engaged in the production of apparently normal sperm lineages, and mature spermatozoa were present within the tubule lumens. In some cases, oocytes were in direct contact with mature spermatozoa within the same seminiferous tubules. The significance of this finding is placed into context with a previous observation of elevated intersexuality in the congeneric species H. americanus collected from specific sites in Canadian waters. The potential mechanism for development of intersex in lobsters, which is probably related to a disrupted signalling to the germinal component of the testis from the decapod androgenic gland, may be an effect of exposure to endocrine disrupting chemicals in the marine environment.

Exposure to the steroid hormone 20-hydroxyecdysone modulates agonistic interactions in male Homarus americanus.

In this study we present evidence that 20-hydroxyecdysone (20E) affects agonistic behavior in male American lobsters and that male and female animals differ in their response to the hormone. Thirty-minute staged fights were conducted between large males exposed either to artificial seawater (ASW) or 20E and small, anosmic opponents. The nephropores of both combatants were blocked. Fights were videotaped and quantitatively analyzed for aggressive, defensive and avoidance behaviors using an ethogram in which behaviors are ranked according to aggressiveness. Unlike female lobsters, exposing male lobsters to 20E did not increase their aggressive behavior; however, there was a marginally significant trend toward an increase in defensive behaviors with a lower aggressive content than in their ASW-exposed counterparts. The opponents of 20E-exposed animals performed significantly more aggressive behaviors than their counterparts. In fights with 20E-exposed animals, the overall aggressive intensity of the fight was increased and the animals performed a greater number of avoidance behaviors. Unlike the effects of 20E on females, where exposure to 20E caused an increase in overall agonistic arousal, males only exhibited a change in frequency of their behaviors. These findings suggest that while 20E affects both males and females in agonistic encounters, the nature of the effect is different for the two sexes.