Properties of pyruvate kinase in the lobster

Pyruvate kinase from the tail muscle and claw muscle of a lobster (H. vulgaris) has been purified by ammonium sulfate precipitation, chromatography on DEAE-cellulose and Sephadex G-200 gel filtration. The overall procedure results in a 200-fold purification with a yield of 70%. The molecular weights of the enzymes were found to be 254 000 ± 10% (tail muscle) and 266 000 ± 10% (claw muscle). Moreover both enzymes have a tetrameric structure with a molecular mass of 65 000 ± 10% for the subunit. Kinetic properties of the enzymes are such that a regulation of anaerobic glycolysis through allosteric modulation of pyruvate kinase activity appears to be non-functional for the two tissues investigated.     

Daily and circadian rhythms of locomotor activity in the American lobster, Homarus americanus

It is widely accepted that American lobsters, Homarus americanus (Milne-Edwards), are nocturnally active. However, the degree to which this rhythm is expressed by different individuals and the underlying causes of lobster activity rhythms, are poorly understood. In order to address these issues we recorded daily patterns of lobster locomotion using two novel techniques. In the first, reed switch assemblies were used to monitor the distance traveled by freely moving lobsters (n=43), each fitted with a small magnet, as they walked around a 1 m diameter racetrack. The advantages of this technique included: (1) lobsters were freely moving; (2) the system could be deployed in laboratory tanks or in the field and; (3) actual distances moved were measured, not just relative activity. The second technique involved placing individual lobsters (n=10) into custom-designed running wheels. This allowed for continuous monitoring of locomotor activity for extended durations (>45 days) under normal light/dark (L/D) cycles, as well as in constant darkness (D/D) and constant light (L/L).Under ambient light conditions lobsters in the racetracks moved an average of 60.1±6.5 m/day in flow-through seawater tanks. Overall, lobsters were significantly more active at night, moving 4.1±0.4 m/h in the dark vs. 1.0±0.2 m/h in the light. However, many of the lobsters moved as much during the day as during the night.Lobsters in the running wheels moved an average of 36.6±11.7 m/day and 80% expressed clear daily rhythms of activity, with a mean periodicity of 24.0±0.1 h under L/D conditions. Under D/D conditions 90% of the animals expressed free-running circadian rhythms with a mean periodicity of 24.2±0.3 h, indicating that this species possesses endogenous rhythmicity. While the running wheel results show that the nocturnal pattern of locomotor activity for this species is strongly influenced by an endogenous circadian clock, the results from the racetracks show that there is remarkable variability in the extent to which they express this pattern under natural conditions.     

Comparison of glycolytic and associated enzyme activities in the claw and tail muscles of the lobster with those of the mantle of Mytilus edulis

In the mussel Mytilus edulis, the activities of phosphoenolpyruvate car?ykinase (PEP-CK) and of malate dehydrogenase (MDH) are greater than those of pyruvate kinase (PK) and lactate dehydrogenase (LDH). The activities of PEP-CK are very low in the lobster (H. vulgaris.) and the ratio of MDH/LDH activities are 0·041 and 1·83 in the tail and the claw muscles respectively. Intracellular L-lactate and L-alanine concentrations suggest a different carbohydrate utilization in the tail and the claw muscle of the lobster. Consistent with this finding is the fact that L-lactate concentration is higher in the tail muscle than in the claw muscle; the opposite is true for L-alanine concentrations.     

Copolymerization of lobster lactate dehydrogenase isoenzymes with pig H4 isoenzyme in vitro

The simple isoenzymatic pattern for LDH in Crustacea and their restrictive ability to copolymerize with mammal isoenzymes indicate early divergence and different evolutionary pressure with respect to control of the catalytic activity.     

An unusual cuticular tumor-like growth on the abdomen of a lobster, Homarus americanus

Tumors are rare in crustaceans, and whereas a few have been reported from the lobster Homarus americanus none have been adequately described. A lobster with an unusual, large, blue-colored tumor-like growth projecting laterally outward from the first abdominal somite was caught off Stonington, Maine, USA. The growth was rugose and covered by a relatively normal appearing cuticle with dispersed focal melanization. The underlying stroma consisted of an internal area of rescaffolded fibrous connective tissue, restructured muscle fibers, few arterioles, and an epidermal area comprised of columnar, highly vacuolated epithelial cells. No infectious pathogens or unusual inclusions were observed with microscopy and no eukaryotic pathogens were detected via molecular sequencing. Given the nature of the histology and the appearance of the growth, we identify the mass as a benign papilliform hamartoma that likely originated as a result of abnormal wound repair possibly initiated around ecdysis. This represents the first tumor-like hamartoma reported from a lobster, and the second hamartoma reported from a crustacean.     

Measurement and Analysis of Extracellular Acid Production to Determine Glycolytic Rate

Extracellular measurement of oxygen consumption and acid production is a simple and powerful way to monitor rates of respiration and glycolysis¹. Both mitochondrial (respiration) and non-mitochondrial (other redox) reactions consume oxygen, but these reactions can be easily distinguished by chemical inhibition of mitochondrial respiration. However, while mitochondrial oxygen consumption is an unambiguous and direct measurement of respiration rate², the same is not true for extracellular acid production and its relationship to glycolytic rate ^3-6. Extracellular acid produced by cells is derived from both lactate, produced by anaerobic glycolysis, and CO₂, produced in the citric acid cycle during respiration. For glycolysis, the conversion of glucose to lactate^- + H⁺ and the export of products into the assay medium is the source of glycolytic acidification. For respiration, the export of CO₂, hydration to H₂CO₃ and dissociation to HCO₃^- + H⁺ is the source of respiratory acidification. The proportions of glycolytic and respiratory acidification depend on the experimental conditions, including cell type and substrate(s) provided, and can range from nearly 100% glycolytic acidification to nearly 100% respiratory acidification ⁶. Here, we demonstrate the data collection and calculation methods needed to determine respiratory and glycolytic contributions to total extracellular acidification by whole cells in culture using C2C12 myoblast cells as a model.     

Effects of near-bed current speeds on the distribution and behaviour of the lobster, Homarus gammarus (L.)

The distribution of the lobster, Homarus gammarus (L.) appears to be limited to areas of the sea bed with rocky outcrops. It seems likely that this limitation operates because lobsters need to be able to avoid currents created by tidal and wave action and that it is size selective. A series of experiments was made with lobsters in a flume tank, to observe the effects of a range of current speeds and substratum types on lobster behaviour. In addition, depth profiles of flow rates were made by Pitot tube measurements to estimate flow rates within the boundary layer immediately above the sea bed. Predictions of the influence of natural water currents on lobster behaviour have been made using available data on near-bed current speeds. The implications of these predictions to fisheries management and reef construction plans are discussed.     

Escape of the cercarial body of Gorgoderina vitelliloba from its anterior tail chamber

Mitchell J. B. and Mason A. R. 1978. Escape of the cercarial body of Gorgoderina vitelliloba from its anterior tail chamber. International Journal for Parasitology8: 193–198. Before the cercarial body of Gorgoderina vitelliloba can encyst in the body of its second intermediate host it must escape from the anterior tail chamber where it has been confined throughout the free living cercarial phase. Escape from the tail chamber is influenced by pH, enzymes, bile salts and by the physical nature of its environment. The cercarial body effects its escape after first becoming activated and then breaking its connection with the tail.     

Changes in oxidative stress parameters and acid phosphatase activity in the pre-regressing and regressing tail of Indian jumping frog Polypedates maculatus (Anura,Rhacophoridae)

Activities of acid phosphatase (normal and Co²⁺-sensitive), superoxide dismutase and catalase and levels of lipid peroxidation, hydrogen peroxide were compared in the tails of tadpoles of stage III, XVIII, XXI and XXIII, respectively, of the Indian Jumping frog Polypedates maculatus. It is noticed that acid phosphatase activity (normal and Co²⁺-sensitive), and levels of lipid peroxidation and hydrogen peroxide increased during tail regression. There is also an increase in the level of superoxide dismutase and catalase in the regressing tail. A positive correlation between activity of acid phosphatase and lipid peroxidation, hydrogen peroxide and lipid peroxidation, acid phosphatase and hydrogen peroxide was noticed in the tail of tadpoles during different developmental stages, suggesting a critical interaction between reactive oxygen species and lysosomal activity during metamorphosis.     

Mechanisms underlying burst generation of the pyloric muscle in the mantis,shrimp, Squilla oratoria

The pyloric constrictor muscles of the stomach in Squilla can generate spikes by synaptic activation via the motor nerve from the stomatogastric ganglion. Spikes are followed by slow depolarizing afterpotentials (DAPs) which lead to sustained depolarization during a burst of spikes. 1. The frequency of rhythmic bursts induced by continuous depolarization is membrane voltage-dependent. A brief depolarizing or hyperpolarizing pulse can trigger or terminate bursts, respectively, in a threshold-dependent manner. 2. The conductance increases during the DAP response. The amplitude of DAP decreases by imposed depolarization, whereas it increases by hyperpolarization. DAPs from successive spikes sum to produce a sustained depolarizing potential capable of firing a burst. 3. The spike and DAP are reduced in amplitude by decreasing [Ca]o, enhanced by Sr2+ or Ba2+ substituted for Ca2+, and blocked by Co2+ or Mn2+. DAPs are selectively blocked by Ni2+, and the spike is followed by a hyperpolarizing afterpotential. 4. The spike and DAP are prolonged by intracellular injection of the Ca2+ chelator EGTA. A hyperpolarizing afterpotential is abolished by EGTA and enhanced by increasing [Ca]o. The DAP is diminished in Na(+)-free saline and reduced by tetrodotoxin. 5. It is concluded that the muscle fiber is endowed with endogenous oscillatory properties and that the oscillatory membrane events result from changes of a voltage- and time-dependent conductance to Ca2+ and Na+ and a Ca2+ activated conductance to K+.     

The anterior tail chamber and survival of Gorgoderina vitelliloba

Mitchell J. B., Mason A. R. and Whalley A. J. S. 1980. The anterior tail chamber and survival of Gorgoderina vitelliloba. International Journal for Parasitology10: 181–182. Survival in pond water of intact cystocercous cercariae of Gorgoderina vitelliloba was significantly better than that of isolated cercarial bodies, although the latter were able to survive in Ringer's Solution. Survival of cercarial bodies in water was improved somewhat when sterility was maintained. It is suggested that the wall of the anterior tail chamber protects the body from the osmotic stress imposed by life in fresh water.     

Changes in blood serum protein levels during the moulting cycle of the lobster, Homarus gammarus (L.)

The blood serum protein level of lobsters kept in a stable environment rose from a mean value of 36.5 μg protein/ml of blood shortly after moulting to about 70 μg/ml in the first 40 % of the moult cycle; in the next 30 % of the cycle there was little change but in the last 30 % the mean blood protein level rose to ≈ 85 μg/ml. It is concluded that the variation in blood protein level between lobsters is too great at any stage of the moulting cycle for the technique to give an indication of the onset of moulting.     

Agonistic interactions between the invasive green crab, Carcinus maenas (Linnaeus) and juvenile American lobster, Homarus americanus (Milne Edwards)

Invasive organisms have the potential for competition with native organisms. In the Southern Gulf of St. Lawrence, juvenile American lobsters have a potential spatial overlap with adult green crabs. Crustaceans use agonistic behaviour to settle disputes, with the larger organism often winning contests for limited resources such as food and shelter. Two experiments were carried out using adult green crabs (53-76 mm carapace width) and juvenile American lobsters (28-57 mm carapace length). The first experiment used a limited food resource. We found that green crabs were the first to the food in all trials, fed in significantly more trials than lobsters and spent a significantly greater proportion of time with the food. The lobsters were only able to displace the green crabs from the food in 2 of 65 attempts. The second experiment was designed to examine shelter competition; unexpectedly some predation by green crabs on lobsters occurred, which allowed us to test hypotheses about how relative size and shelter use affect predation. Green crabs captured and consumed juvenile lobsters in 6 of 11 trials. The lobsters that survived spent significantly more time in shelter. There was no clear relationship between shelter use and size of lobster. The lobsters that were larger in relation to the green crabs suffered a higher rate of predation, which we believe was due to more conspicuous activity and less use of shelter. It appears that green crabs have the potential to negatively impact native juvenile lobster.     

Ventilation and oxygen consumption in the hagfish, Myxine glutinosa L.

Ventilation was measured directly in the hagfish, Myxine glutinosa L., by means of an electro-magnetic blood flowmeter. Ventilatory flow and frequency increased from 0.86 ± 0.27 ml·min^?, and 18.2 ± 5.1·min^?, respectively, at 7°C to 1.70 ± 0.20 ml·min^?, and 70.1 ± 9.5·min^? at 15 ·C.Standard oxygen consumption,V?O₂, was measured in non-buried hagfish. V?O₂ was 0.57 ± 0.17μl O₂·g^?1·min^?1 at 7°C, and 0.85 ± 0.12μl O₂·g^?1·min^?1 at 15°C.     

Histopathology of Aerococcus viridans var. homari infection (Gaffkemia) in the lobster, Homarus americanus, and a comparison with histological reactions to a gram-negative species, Pseudomonas perolens

Histological response of lobsters to injection of Aerococcus viridans var. homari, cause of gaffkemia, was followed over a 14-day period. Salient features in infected lobsters, Homarus americanus, were: aggregations of hemocytes occurring in hemal spaces throughout the tissues and increasing in number and size with time; the early phagocytosis of bacteria by the system of fixed phagocytes (FPs) present in hemal spaces of the hepatopancreas; and premature release of differentiating hemocytes from the hemopoietic tissue, so that by 14 days that tissue consisted mainly of large stem cells. Mass release of differentiating hemocytes presumably occurred to replace hemocytes lost from the circulation by their incorporation into aggregations or by lysis of individual cells ruptured through the pressure of phagocytized bacteria that were multiplying in them. Bacteria and their remains were present in FPs at 2 days but not visible in single or aggregated hemocytes until 6 days, when free bacteria were also present in the hemolymph. By 6 days, all bacteria, whether phagocytized or free, appeared normal and were surrounded by nonstaining halos that extended well beyond the stainable capsular material. As predicted earlier in physiological studies, gaffkemia is a nontoxic, noninvasive bacteremia. There was hemal stasis and consequent injury in the antennal gland due to free and aggregated hemocytes that occluded hemal spaces of that organ, but other tissues and organs appeared normal except for depletion of glycogen. Aggregations of hemocytes were present in lobsters 2 and 12 days after injection of a nonpathogenic, Gram-negative bacterium, Pseudomonas perolens. Unlike the case with gaffkemia, necrotic hemocytes were common in the aggregations, presumably in response to damage by endotoxin. A further difference was that aggregations were common in the heart of P. perolens-injected lobsters but rare in the heart of gaffkemic lobsters. Bacteria were not seen in hemolymph, hemocytes, or other cells of P. perolens-injected lobsters.     

The claw closer muscle of Neohelice granulata (Grapsoidea,Varunidae): a morphological and histochemical study

Longo, M.V., Goldemberg, A.L. and Díaz, A.O. 2011. The claw closer muscle of Neohelice granulata (Grapsoidea, Varunidae): a morphological and histochemical study. —Acta Zoologica (Stockholm) 92 : 126–133. The claw closer muscle of Neohelice granulata was studied according to histological, histochemical, and morphometrical criteria. Adult male crabs in intermoult stage were collected from Mar Chiquita Lagoon (Buenos Aires, Argentina). Muscle fibers show evident striations and oval‐elongated nuclei with loose chromatin. The loose connective tissue among muscle fibers consists of cells and fibers embedded in an amorphous substance. Muscle histochemistry reveals two slow fiber types: ‘A’ and ‘B’. Prevailing A fibers are larger, and they usually show, with respect to B type, a weaker reaction to whole techniques. Fibers with short (SS), intermediate (IS), and long sarcomeres (LS) appear in the claw closer muscle, being the LS fibers predominant. Concluding, the histochemical and morphometrical characteristics of the claw closer muscle fibers of N. granulata are indicative of slow fibers. The slow A type (low resistant to fatigue) prevails.     

The external morphology and distribution of cuticular hair organs on the claws of the American lobster, Homarus Americanus (Milne-Edwards)

Four classes of microscopic cuticular hair organs were found on the chelae of Homarus americanus (Milne-Edwards). Type I and Type II organs possess long single sensillae, 30–60 μm and 70–130 μm, respectively. Type III organs are toroid bumps 20–30 μm in diameter, with a small tuft of fibers projecting from the center. Type IV organs are small conical hairs ≈ 1 μm in length.Type IV organs were uniformly distributed over the claws. The distributions of Types I, II, and III organs were analysed through multivariate analysis of variance (MANOVA) with respect to claw, side, and area. Type I organs showed significance of the three-way interaction only. Type II organs showed significance of side, area, and the claw by area interaction. Type III organs showed significance of area, and all interactions. Changes in number and density of Types I, II and III organs, and in number of Type IV organs, were found with growth.     

Lobster (Homarus americanus) hemocytes: Classification,differential counts,and associated agglutinin activity

Four hemocyte types were recognized in the lobster based on size and refractile nature of the granules, the ratios of cytoplasm to nucleus, and Giemsa stain characteristics. Two hyaline types were designated as prohyalocytes (1.8%) and hyalocytes (64.2%), and two granular types were termed eosinophilic granulocytes (12.2%) and chromophobic granulocytes (21.9%). There was no significant difference in the percentages of the different hemocyte types (differential hemocyte counts) between sexes, but hyalocyte and eosinophilic granulocyte percentages varied significantly between populations of lobsters. The data suggested that the difference in agglutinin activity (HA) between lobsters with the same total hemocyte numbers was due to activity associated with fixed hemocytes or quantitative differences in HA activity associated with one or more hemocyte types, rather than an increase in the percentage of any one particular type in circulation.