Amino acids in haemolymph, single fibres and whole muscle from the claw of freshwater crayfish acclimated to different osmotic environments

The concentrations of free amino acids were measured in whole claw muscle, single fibres and haemolymph of Australian freshwater crayfish, Cherax destructor, during the intermoult stage. The average total pool of amino acids in short-sarcomere fibres (179 mmol kg(-1)) was 60% greater than in long-sarcomere fibres, due to higher concentrations of alanine, cysteine, glutamate, leucine and proline. The two fibre types exhibited differences in the banding pattern of the isoforms of troponin using gel electrophoresis. The average pool of amino acids in haemolymph was 2.7 mmol kg(-1). Cherax has symmetrical claws and the total pool of amino acids from whole muscles (approx. 79 mmol kg(-1)) was similar in left and right claw muscles. In animals acclimated to osmotic environments between 0 and 220 mOsm, the osmotic pressure of the haemolymph increased from 356 to 496 mOsm, but no systematic changes were observed in the amino acid profiles of muscles or haemolymph. The major findings were that (a) concentrations of amino acids differed between the two major fibre types in claw muscle and (b) amino acids in the muscle fibres did not play a major part in intracellular osmoregulation in Cherax, suggesting this species is an anisosmotic regulator.     

Calcium,phosphorus and adenylate levels and Na(+)-K(+)-ATPase activities of prawn,Macrobrachium nipponense,during the moult cycle

Changes in calcium and phosphorus concentrations, adenylate (AMP, ADP and ATP) levels, and ratios and ATPase activities of Macrobrachium nipponense were investigated during the moult cycle. Ca level in the exoskeleton was lowest in early postmoult (stage A), increasing at stages B and through intermoult (stage C) and peaking in premoult (stage D1 and D2). The P concentrations in the exoskeleton and muscle in late premoult and early postmoult stages were higher than those at other moult stages, and were lowest in the intermoult. Muscle adenylate energy charge (AEC) changed with moult stages, and was in agreement with the change in inorganic P level in the muscle. AEC may be a direct indicator of energy metabolic activity during the moult cycle. ATP/ADP and ATP/AMP ratios in premoult and postmoult stages were higher than that in intermoult stage. Na(+)-K(+)-ATPase activities of gills, muscles and hepatopancreatic of prawns were higher in early postmoult and late premoult animals, whereas they were lower in late postmoult, intermoult and early premoult animals. Gill residual ATPase activity was significantly higher in postmoult animals, while the peak value of hepatopancreatic residual ATPase activity appeared in intermoult stage.     

Haemolymph constituents and osmolality as functions of moult stage,body weight,and feeding status in marron,Cherax cainii () and yabbies,Cherax destructor (Clark, 1936)

The study investigates the change in osmolality and haemolymph constituents in marron Cherax cainii and yabbies Cherax destructor associated with moult stages, body weights and their feeding status. A total of 582 haemolymph samples from 5 moult stages (postmoult-AB, intermoult-C, and premoult stages – D₀, D₁, D₂), two body weight classes (2–15 g and 61–75 g) and nutritional status were used for analysis of osmolality, protein, glucose, and ionic concentrations of potassium and chloride following the standard biochemical procedures. The haemolymph protein, glucose, potassium and chloride levels were highest at intermoult and early premoult stages, and lowest at postmoult in both crayfish species. Except protein, no significant differences were seen in analyzed parameters between various weight classes and two species. Haemolymph osmolality, protein and glucose were significantly higher in fed crayfish, whereas no variations in haemolymph potassium and chloride concentrations were observed between the fed and unfed crayfish. Maximum osmolality was recorded at 7–8 h after feeding in both crayfish species. The results showed that the biochemical changes in the haemolymph of marron and yabbies are related to moult stages, body weight and feeding and thus can be used as tools for determining suitable diets.     

The immune response of the white shrimp Litopenaeus vannamei and its susceptibility to Vibrio infection in relation with the moult cycle

The white shrimp Litopenaeus vannamei (8.0-14.4 g) was examined for haemocyte count, phenoloxidase activity, respiratory burst (release of superoxide anion), phagocytic activity, and clearance efficiency to the pathogen Vibrio alginolyticus in relation with moult cycle (postmoult, A, B; intermoult, C; premoult, D(0)/D(1)D(2)/D(3)). Granular cells were the highest at C and D(0)/D(1)stage, and the lowest at A stage. Hyaline cells and THC (total haemocyte count) were higher at C stage, but lower at postmoult stages. Phenoloxidase activity was the highest at C stage, and the lowest at A stage. Respiratory burst was lower at A stage. Phagocytic activity of shrimps against V. alginolyticus decreased significantly at postmoult and premoult stages. Additionally, the clearance efficiency of shrimps to V. alginolyticus was significantly lower for shrimps at A stage than those at C stage. In another experiment, L. vannamei at different moult stages were injected with tryptic soy broth (TSB)-grown V. alginolyticus (1x10(5)cfu shrimp(-1)) and then held in 34% seawater. After 10 h, the mortality of V. alginolyticus-injected shrimps was significantly higher for shrimps at postmoult stage than those at intermoult stage. Over 48-120 h, the mortality of V. alginolyticus-injected shrimps was 50.0%, 33.3% and 40.0% at postmoult, intermoult and premoult stage, respectively. It is concluded that L. vannamei showed a decrease in resistance at A stage through a reduction of its haemocyte count, phenoloxidase activity, respiratory burst, phagocytic activity and clearance efficiency against V. alginolyticus.     

Effects of seasonality and moult cycle on the proliferation of nerve cells and on the labelling of ecdysone receptors in an estuarine crab

Decapod crustaceans show proliferation of the nerve cells in the olfactory lobe throughout their lives. However, the regulation of this process is still poorly understood, since it may vary with endogenous and exogenous factors. The objective of the present investigation was to quantify the proliferation of nerve cells and number of nerve cells with ecdysone receptors in the clusters of the central olfactory system in Neohelice granulata, according to moult stages and in different seasons (summer and winter). Three injections of bromodeoxyuridine (BrdU) were administered to the crabs. Brains were sectioned by microtome and fixed on slides for immunohistochemistry with anti-BrdU and anti-EcR antibodies. The proliferation of nerve cells was higher in winter than in summer, probably because in winter the crabs do not breed and the premoult and postmoult periods are longer. Crabs in postmoult exhibited more BrdU-labelled cells than crabs in premoult or intermoult in winter, because of a greater number of mitoses related to an increase in body size and addition of olfactory receptor neurons. The number of EcR-labelled cells was higher in premoult than in postmoult or intermoult in winter. The proliferation of nerve cells is regulated seasonally and according to moult stages.     

Tissue accumulation and the moult cycle in juveniles of the Australian freshwater crayfish Cherax destructor

1. This paper investigates moult stage and size-specific changes in whole body composition during growth in juvenile crayfish in order to better describe the nature of growth and energy use. 2. Composition is described in terms of moult stage and size-specific wet, dry and ash-free dry weight, water, carbon, protein nitrogen, non-protein nitrogen, ash and energy. Dry weight and ash-free dry weight (AFDW) peaked in the middle and in the later stages of premoult. Both peaks were about 2–2.3 times postmoult weight. Losses in tissue weight during ecdysis were substantial in the smallest crayfish but declined with size. Water was taken up between late premoult and early postmoult. Tissue accumulation occurred primarily between B and D₁.₃, with further weight gain largely the result of fluid uptake. Ash increased immediately postmoult with a major peak occurring during intermoult. Mean organic carbon varied between 33 and 35.5% of the body and 49% of the exuvia. Chitin varied between 9 and 17% of body AFDW and made up about 50.5% of the exuvia. Protein content varied between 47 and 62% of body AFDW and about 25% of the exuvia. Carbon, chitin and protein were not affected by moult stage but protein declined with ocular carapace length (OCL) in larger crayfish. Mean caloric content varied between 19 and 22 J mg-¹ AFDW depending on size and moult stage. Caloric content increased with OCL during premoult and early intermoult then declined with size until part of the way through premoult. 3. Relationships between protein, chitin and remaining carbon (organic carbon minus chitin) were examined. It is suggested that protein and some carbon are catabolized during the moulting process, possibly to fuel metabolism. Models are presented showing changes in proximate composition over the moult cycle for two sizes of crayfish, and tissue and energy accumulation and loss over a series of moult cycles and sizes from 3.1 mm to 17 mm OCL.     

Moult cycle and its chronology in Mysis mixta and Neomysis integer (Crustacea, Mysidacea): implications for growth assessment

The moult-staging technique was used to determine the main moult stages (MS) and their duration in laboratory-reared juveniles and subadults of Mysis mixta and Neomysis integer. The relative duration of each stage was similar for both species with premoult occupying the major part of the moult cycle (51% and 44% for M. mixta and N. integer, respectively), followed by postmoult (26% and 34%) and intermoult (23% and 22%). Effects of temperature and feeding regimes on the chronology of the moult cycle were investigated. When the duration of the moult cycle (MCD) was extended by manipulating the feeding regime, animals prolonged their late postmoult and early premoult stages. At 12 and 5 °C, no specific moult stage varied in relative duration as long as food supply was high. Field application of moult staging for growth assessment was tested using wild-caught M. mixta. The MCD estimated via moulting experiments was compared to that obtained by analyzing moult stage distribution combined with the experimentally obtained data on stage duration. Close correspondence between two methods of MCD assessment was observed with moult-staging technique being particularly useful in situations when conducting of experiments immediately after collection is not feasible.     

Moult cycle and morphogenesis inHyas araneus larvae (decapoda,majidae), reared in the laboratory

Moult cycle and morphogenesis in larval instars (zoea I, zoea II, megalopa) of the spider crabHyas araneus (L.) were studied in the laboratory. Changes in the epidermis and cuticle were documented photographically at daily intervals to characterize the stages of the moult cycle. Stage A (early postmoult) is a very short period during which the larva takes up water. During late postmoult (B) and intermoult (C) the endocuticle is secreted, and there is conspicuous epidermal tissue condensation and growth. The onset of early premoult (D₀) is characterized by epidermal apolysis, occurring first at the bases of the setae in the telson of zoeal instars or in the rostrum of the megalopa, respectively. Intermediate premoult (D₁) is the main period of morphogenesis, in particular of setogenesis: in the setae of the zoeal telson and carapace there is invagination or (in the zoea II) degeneration of epidermal tissues. Formation of new setae in the interior of epidermal tubules was observed in zoeal maxillipeds and in the antennae of the zoea II and megalopa instars. During late premoult (Stages D_2–4) part of the new cuticle is secreted, and the results of morphogenesis become clearly visible. For technical reasons (rigid exoskeleton) only a preliminary account of the moult cycle in the megalopa can be given. A time schedule is suggested for the stages of the moult cycle. It is estimated that postmoult (A–B) takes ca 9 to 15 % of total instar duration, intermoult (C) ca 22 to 37 %, and premoult (D) ca 48 to 69 %. There is an increasing trend of relative portions of time (% of total instar duration) from instar to instar in Stages A–C (mainly in the latter) and a decreasing trend in Stage D (mainly in D₀ and D_2–4).     

Two fast-type fibers in claw closer and abdominal deep muscles of the Australian freshwater crustacean, Cherax destructor, differ in Ca2+ sensitivity and troponin-I isoforms

One type of fast fiber and two types of slow (slow-twitch, S1 and slow-tonic, S2) fibers are found in decapod crustacean skeletal muscles that differ in contractile properties and myofibrillar protein isoform compositions. In this study the structural characteristics, protein isoform compositions, and Ca2+-activation properties of fast fibers in the claw closer (F1) and abdominal deep flexor (F2) muscles of Cherax destructor were analyzed. For comparison, myofibrillar protein isoform compositions of slow (long-sarcomere) fibers from claw and abdomen were also determined; our results indicate that the slow fibers in the claw closer were the slow-twitch (S1) type and those in the abdominal superficial flexor were primarily slow-tonic (S2) type. F1 fibers had shorter resting sarcomere lengths (2.93 microm in unstretched fibers and 3.06 microm in stretched fibers) and smaller fiber diameter (256 microm) than F2 fibers (sarcomere lengths 3.48 microm in unstretched and 3.46 microm in stretched; 747 microm diameter). Moreover, F1 fibers showed a narrower range in sarcomere lengths than F2 fibers (2.81 to 3.28 microm vs. 2.47 to 4.05 micro m in unstretched fibers). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting showed that the fast fibers from claw and abdomen differed in troponin-I composition; F1 fibers expressed two isoforms of troponin-I (TnI1 and TnI2) in approximately equal amounts, whereas F2 fibers expressed primarily TnI3 and lower levels of TnI1. F1 fibers were more sensitive to Ca2+, as shown by higher pCa values at threshold activation (pCa(10)=6.50+/-0.07) and at 50% maximum force (pCa(50)=6.43+/-0.07) than F2 fibers (pCa(10)=6.12+/-0.04 and pCa(50)=5.88+/-0.03, respectively). F1 fibers also had a greater degree of co-operativity in Ca2+ activation, as shown by a higher maximum slope of the force-pCa curve (n(Ca)=12.98+/-2.27 vs. 4.34+/-0.64). These data indicate that there is a greater fast fiber-type diversity in crustacean muscles than was previously supposed. Moreover, the differences in activation properties suggest that the TnI isoform composition influences the Ca2+ sensitivity of the contractile mechanism.     

Effects of Removal of Muscle Receptor Organ Input on the Temporal Structure of Non-Giant Swimming Cycles in the Crayfish, Cherax Destructor

Most studies of the muscle receptor organs (MROs) of decapod crustaceans have focused on their role in local reflex loops. This may not be their only function. We examine their involvement in the regulation of non-giant swimming cycles by removing stretch receptor (SR) input from the MROs in abdominal segments 2-5 of the crayfish Cherax destructor . SR input was left intact in two control groups, one of which had sham surgery and the other no surgery at all. We recorded electromyograms (EMGs) from selected uropod muscles during tailflipping in sequences of non-giant swimming in tethered animals. The removal of SR input had a significant effect. The opener muscle period was shorter in the experimental group than in either of the control groups. This suggests that by using SR afference, crayfish sacrifice speed for increased control of the swimming movement.     

Effects of Removal of Muscle Receptor Organ Input on the Temporal Structure of Non-Giant Swimming Cycles in the Crayfish,Cherax Destructor

Most studies of the muscle receptor organs (MROs) of decapod crustaceans have focused on their role in local reflex loops. This may not be their only function. We examine their involvement in the regulation of non-giant swimming cycles by removing stretch receptor (SR) input from the MROs in abdominal segments 2-5 of the crayfish Cherax destructor. SR input was left intact in two control groups, one of which had sham surgery and the other no surgery at all. We recorded electromyograms (EMGs) from selected uropod muscles during tailflipping in sequences of non-giant swimming in tethered animals. The removal of SR input had a significant effect. The opener muscle period was shorter in the experimental group than in either of the control groups. This suggests that by using SR afference, crayfish sacrifice speed for increased control of the swimming movement.     

Osmoregulatory capacity,health status and growth as functions of moult stages from various weight classes in marron (Cherax cainii) and yabbies (Cherax destructor)

The study investigated the relationship between five moult stages and the osmoregulatory capacities (OC), moisture content of hepatopancreas (HM%), hepatosomatic indices (HI_wet, HI_dry) and growth of 6 weight classes of 235 marron (Cherax cainii) and 235 yabbies (Cherax destructor) under laboratory conditions. In each moult stage, OC increased linearly with the increase in wet body weight. Intermoult stage C showed the highest OC significantly lower than premoult stages D₀, D₁, D₂ and postmoult stage AB in every weight class in both crayfish. HM% decreased from AB to D₂, while both HI_wet and HI_dry reduced from C to D₂. Percentage dry matter of whole body carcass was highest in stage C and lowest during the AB stage with a significant difference between the two species. Larger than 15-g marron and yabbies differ with each other in OC, HI_wet, and HI_dry, moult intervals and growth rates.     

The moult cycle of the terrestrial isopod Armadillo officinalis Duméril, 1816 (Crustacea: Isopoda: Oniscidea)

The present work focuses on the moult cycle of Armadillo officinalis. For a 100‐day period, 134 animals were observed and routinely examined with the aim of detecting distinctive morphological characters in the several stages and substages of the moult cycle and of disclosing their duration. Statistical tests and Poisson regression models with robust standard errors were used to investigate differences and relationships between moult and the size and gender of the animals. The appearance of the calcium carbonate deposits on the pereon sternites during the premoult stage was documented in detail, and three main substages were identified. The average duration of the premoult and of the biphasic ecdysis was about 12 and 1.5 days, respectively. This observation period, however, did not allow to establish a determined average duration of the intermoult stage, which was extremely variable. This stage lasted for 2 months or more in most of the cases observed, but about 1‐month‐long intermoult stages were also recorded. No statistically significant association was found between the number of moults and gender and size of the animals.     

Endocrine cells in the gut of the shore crab Carcinus maenas immunoreactive to crustacean hyperglycaemic hormone and its precursor-related peptide

The distribution and morphology of gut endocrine cells, which are immunoreactive to crustacean hyperglycaemic hormone (CHH) and the corresponding precursor-related peptide (CPRP), have been described in the shore crab Carcinus maenas. The cells are uniquely distributed throughout the fore- and hindgut, but were never observed in the midgut or associated caeca. Expression of CHH and CPRP in the gut endocrine cells is generally restricted to premoult, although small numbers of immunoreactive cells were observed in intermoult and postmoult. A notable feature of the distribution of these slender cells was that, whilst they are distributed evenly over much of the fore- and hindgut, all extrinsic and intrinsic muscles of the gastric and pyloric stomach examined were surrounded by a ring(s) of cells, suggesting a mechanoreceptive function. Ultrastructural studies revealed that these cells contain numerous immunopositive, electron-dense granules. This suggests that they are "paraneurones", which secrete CHH and CPRP into the haemolymph during ecdysis, accounting for the ecdysial surge in CHH, which is implicated in water uptake and swelling prior to ecdysis.     

Metabolic changes in the African fruit beetle,Pachnoda sinuata,during starvation

Specimens of the fruit beetle Pachnoda sinuata were starved for up to 30 days. The weight of the beetles declined consistently throughout the starvation period. Concentrations of carbohydrates and alanine in flight muscles, fat body and haemolymph decreased rapidly after onset of starvation, while the concentration of proline remained high. Whereas the lipid concentrations in the haemolymph did not change significantly upon starvation, the lipid content in flight muscles and fat body decreased significantly.Beetles that had been starved for 14 days responded to injection of Mem-CC, the endogenous neuropeptide from its corpora cardiaca, with hyperprolinaemia and a decrease in the alanine level, but no such effect was monitored after prolonged starvation of 28 days. Regardless of the period of starvation, Mem-CC injection could not cause hypertrehalosaemia or hyperlipaemia, although carbohydrates were increased in fed beetles after injection.Flight ability of beetles that had been starved for 15 or 30 days was apparently not impaired. During such periods, beetles used proline exclusively as fuel for flight as evidenced by the increase in the level of alanine in the haemolymph and decrease of the level of proline; the concentrations of carbohydrates and lipids remained unchanged.Activities of malic enzyme and alanine aminotransferase (enzymes involved in transamination in proline metabolism), glyceraldehyde-3-phosphate dehydrogenase (enzyme of glycolysis), 3-hydroxyacyl-CoA dehydrogenase (enzyme of beta-oxidation of fatty acids) and of malate dehydrogenase (enzyme of Krebs cycle) were measured in fat body and flight muscles. In flight muscle tissue the maximum activity of NAD(+)-dependent malic enzyme increased, while that of glyceraldehyde-3-phosphate dehydrogenase decreased during starvation, and malate dehydrogenase, 3-hydroxyacyl-CoA dehydrogenase and alanine aminotransferase were unchanged. In fat body tissue, activities of NADP(+)-dependent malic enzyme and 3-hydroxyacyl-CoA dehydrogenase increased during food deprivation and activities of glyceraldehyde-3-phosphate dehydrogenase, malate dehydrogenase and alanine aminotransferase remained unchanged.     

Ammonia excretion of the sand-shrimpCrangon crangon (L.) during the moult cycle

Summary Ammonia excretion in the shrimpCrangon crangon (L.) shows a cyclical pattern during the moult cycle. At the early premoult (stage D₀) there is a 7 to 17% increase over that characteristic at intermoult; at late premoult (stage D₂) excretion decreases to a minimum; immediately after ecdysis, the excretion rate is 1.5 to 2.6 times higher than at the intermoult stage (Table 1).These variations appear to be correlated with protein breakdown and protein synthesis, and possibly with periods of high metabolic activity (growth resulting from cell multiplication).Ammonia excretion of animals in standing water is higher than of those in running sea-water. Excretion might possibly be influenced by a semi-lunar rhythm.     

Effects of reduced salinities on development and bioenergetics of early larval shore crab, Carcinus maenas

The shore crab, Carcinus maenas L. (Portunidae), is a coastal and estuarine species, which can live and reproduce under brackish water conditions; freshly hatched larvae have been observed in the field at salinities below 15‰. In the present laboratory study, the tolerance of hypo-osmotic stress was experimentally investigated in early larvae of a marine (North Sea) population of C. maenas reared at four different salinities (15, 20, 25, 32‰). Two and 4 days after hatching, the Zoea I larvae were moult-staged microscopically, and their rates of respiration and growth (changes in dry weight, W, carbon, C, nitrogen, N, and hydrogen, H) were measured. Survival and development were monitored until the megalopa was reached: 15‰ did not allow for development beyond the first zoeal stage, while metamorphosis to the megalopa was reached at salinities ≥20‰. At 20‰, development was significantly delayed and mortality enhanced as compared with 25 and 32‰. Rates of growth and respiration decreased during exposure to reduced salinities ≤25‰. Hence, the suppression of growth could not be explained as a consequence of enhanced metabolic losses per larva. Instead, a partial C budget indicates that the Zoea I larvae suffered from decreased capabilities of assimilating ingested and subsequently converting assimilated matter to tissue growth. Net growth efficiency (K₂, C-based) was at 25 and 32‰ initially high (>60% during the postmoult and intermoult stages of the Zoea I moult cycle), but decreased during the later stages (down to ≤30% in premoult). An inverse pattern of C partitioning was observed at ≤20‰, with initially low K₂ values (≤21% during the first 2 days of the moult cycle), and a later increase (up to ≥46% in premoult). Thus, larval growth was initially suppressed under conditions of reduced salinity, but this was later (during premoult) partially compensated for by an increase in C assimilation and K₂. Our observations indicate that Zoea I shore crab larvae react during the late stages of their moulting cycle less sensitively against reduced salinities than during postmoult and intermoult. This suggests that the transition between moult cycle stages C and D₀ may be a critical point for effects of hypo-osmotic stress, similarly as already known in relation to effects of nutritional stress. Negative effects were found also when freshly hatched Zoea I shore crab larvae were exposed only transitorily (for 24–72 h) to 20‰, with significantly lower rates of survival, development, growth, respiration, and K₂. These effects increased with increasing duration of initial exposure to reduced salinity.     

Whole body and skeletal muscle glutamine metabolism in healthy subjects

We measured glutamine kinetics using L-[5-15N]glutamine and L-[ring-2H5]phenylalanine infusions in healthy subjects in the postabsorptive state and during ingestion of an amino acid mixture that included glutamine, alone or with additional glucose. Ingestion of the amino acid mixture increased arterial glutamine concentrations by approximately 20% (not by 30%; P < 0.05), irrespective of the presence or absence of glucose. Muscle free glutamine concentrations remained unchanged during ingestion of amino acids alone but decreased from 21.0 +/- 1.0 to 16.4 +/- 1.6 mmol/l (P < 0.05) during simultaneous ingestion of glucose due to a decrease in intramuscular release from protein breakdown and glutamine synthesis (0.82 +/- 0.10 vs. 0.59 +/- 0.06 micromol x 100 ml leg(-1) x min(-1); P < 0.05). In both protocols, muscle glutamine inward and outward transport and muscle glutamine utilization for protein synthesis increased during amino acid ingestion; leg glutamine net balance remained unchanged. In summary, ingestion of an amino acid mixture that includes glutamine increases glutamine availability and uptake by skeletal muscle in healthy subjects without causing an increase in the intramuscular free glutamine pool. Simultaneous ingestion of glucose diminishes the intramuscular glutamine concentration despite increased glutamine availability in the blood due to decreased glutamine production.     

Calcium balance in premoult and post-moult Gammarus pulex (Amphipoda)

SUMMARY. Adult Gammarus pulex lose about 42% body calcium into solution over a 2–3-day period preceding the moult. A further 54% body calcium is lost with the exuviae, leaving c . 4% in the newly moulted animal.
Nearly all of the body calcium in a stage A (post moult) animal is contained in the hepatopancreas and a transient increase in the calcium content of this tissue is seen in some individuals. The haemolymph calcium compartment is heavily depleted at this stage. In stage B animals the hepatopancreas calcium level has returned to, or even fallen below, the intermoult level while the haemolymph calcium concentration remains lower than in intermoult animals.
Animals switch from a strongly negative calcium balance to a state of rapid calcium uptake immediately following moult. Uptake at 10°C proceeds at a rate of between 3 and 10 μmol g⁻¹ h⁻¹ depending on the calcium concentration of the external medium. The lower rate is found in starved animals in 0.1 mM calcium and this concentration is probably close to a minimum for the satisfactory restoration of body calcium. Recalcification is completed in 10–14 days in 0.1 mM calcium and is apparently enhanced by the presence of food in the form of a portion of oak leaf together with the cast exuviae. In 1.0 mM calcium the recalcification period is shortened to 3–4 days. This is considerably longer than the time recorded for French populations of this species (Vincent, 1969), although reasons for this are offered.     

Lactic acid formation in crustaceans and the liver function of the midgut gland questioned.

1. The possibility of the midgut gland of the crustacean (Cherax destructor) functioning as a liver has been investigated. 2. Seven species of crustaceans accumulate lactic acid in the haemolymph when exercised. The rate of disappearance of lactate in Homarus gammarus and in C. destructor is very slow when compared with man. 3. In the midgut gland of C. destructor no firm evidence was obtained for gluconeogenesis from lactate and for ketogenesis from fatty acids. 4. It is concluded that there is at present no justification for the common practice of calling the midgut gland an hepatopancreas.