ALBERT

Notes: Abstract The distribution and fate of14C-proline were investigated in immature tiger prawns,Penaeus esculentus Haswell, collected in Moreton Bay, Cleveland, Australia, by trawling during 1986/1987. Initially the prawns were fed14C-proline in food pellets to follow the pathway of proline absorption and distribution in the body.14C-proline was also injected directly into the prawn to provide sufficient tracer to follow the incorporation of14C into other amino acids and into proteins. A comparison was made of the metabolism of injected14C-proline over 48 h in prawns that had been fed and those that had been starved for 10 d. Free amino acids (FAA) in the muscle and protein-bound amino acids were analysed separately. Labelled proline was completely absorbed and distributed within the body 3 h after ingestion, about 80% being in the tissues, mostly in muscle. There was no significant difference between the total CO2 output in fed and starved prawns, but the latter metabolised about twice the amount of labelled proline over 48 h. At this time, in abdominal muscle of fed prawns, about 95% of the total muscle label was in the FAA; of the label in the FAA, 78% was proline and 18% glutamic acid, with the remainder in hydroxyproline, aspartic acid, glutamine, alanine and Kreb's cycle intermediates. In the starved prawns, proline was 58%, glutamic acid 24%, with correspondingly higher amounts in the other compounds. In the muscle protein, the distribution of label was similar in fed and starved prawns, with 72 to 74% as proline. The experiments showed that proline is not very labile in the tiger prawn and its rate of synthesis is slow. It does not appear to be an important source of energy as in some insects and cephalopods, but during starvation is only slowly oxidised for energy.

Notes: Abstract Penaeus esculentus Haswell were collected from Moreton Bay, Queensland, Australia, by trawling during 1987–1988. After standardisation for nutritional state and moult stage (C to D0), prawns were fed one of three 14C labelled lipids: a non-essential fatty acid (FA: palmitic acid, 16:0); an essential polyunsaturated fatty acid (PUFA: eicosapentaenoic acid, 20:5n-3); and the essential sterol, cholesterol. The diet was not deficient in any of these lipids. Experiments were run for 24 and 72 h, the rate of production and radioactivity of CO2 was determined, and at the end of each experiment lipid classes of the neutral (NL) and polar lipid (PL) fractions of the digestive gland, abdominal muscle and integument were separated and the 14C counted. Approximately 34% of the 14C16:0 and 14C20:5n-3 was oxidised to CO2 at similar rates up to 72 h, both reacing a peak at 12 h; cholesterol was not oxidised to CO2. The distribution in the tissues of 14C from all three labels was similar: digestive gland 〉 muscle 〉 remainder 〉 blood 〉 gills 〉 proventriculus 〉 hindgut. This was not due to mass of the tissue or its lipid content. Most of the label from the FA was in PL (digestive gland 〉 50%, muscle and integument 〉 80%). In the NL, most of the label was in free FA; in the PL classes, 14C was predominant in phosphatidylcholine (PC), especially in the muscle and integument. The data indicate that the digestive gland is a major site of lipid synthesis, as well as assimilation and storage. The distribution of the 14C20:5n-3 label differed appreciably from that of 14C16:0 only in the PL fraction, where it was more evenly distributed. It was concluded that, when in excess, the fate of this essential PUFA is similar to that of non-essential FA. Labelled cholesterol was distributed readily through the tissues, but appeared to be mostly retained as such.

Notes: Abstract Neutral lipid and phospholipid fractions and their component lipid classes in the digestive gland, abdominal muscle, epidermis and cuticle ofPenaeus esculentus Haswell were analysed and compared during the moulting cycle and during starvation. The prawns were collected from Moreton Bay, Queensland, Australia, by trawling during 1985–1987, and were fed on a standard, semi-purified diet. The digestive gland appears to be a major site of lipid synthesis, storage and mobilisation in preparation for moulting. Neutral lipid, 59 to 80% of which was triacylglycerol, was the larger fraction. It accumulated during early premoult, mainly due to the increase in triacylglycerol. The digestive gland contained only 18% of the total body lipid, or 8% of body lipid as triacylglycerol. Thus, the reserve lipid available for energy production is very small. Digestive gland triacylglycerol was markedly depleted after 4 d starvation and was almost completely absent after 8 d. In the other tissues, the major fraction was phospholipid, of which over 50% was phosphatidylcholine and up to 20% phoshatidylethanolamine; cholesterol was the major class in the neutral lipid fraction and appeared to be very stable. Most of this lipid was probably a component of cellular membranes. The lipid composition of muscle changed very little during the moulting cycle: total lipid levels in the epidermis were high in late premoult and early postmoult, when new cuticle is being secreted, but the proportions of the component lipids were closely similar. Cuticle lipid, together with other major components, was resorbed from the old cuticle prior to ecdysis, but the cuticle phospholipids appeared to be labile at all moult stages. The total of all lipids in fedP. esculentus was about 3.6% dry weight, of which about 70% was phospholipid. Earlier research had shown that when digestive gland lipid is exhausted after a short period of starvation, muscle is metabolised for energy. The present research showed that in the remaining muscle only about 13% of lipid was lost after 21 d starvation, mostly as phosphatidylcholine. This is in keeping with the need to maintain this tissue in a functional state. In contrast, epidermal lipid levels were markedly reduced after only 4 d starvation and the proportions of phospholipids changed significantly. This sensitivity of the cuticle lipids to starvation may be the cause of delayed moulting, which is characteristic of poor nutrition.

Notes: Abstract Penaeus semisulcatus de Haan adult gravid females, eggs and non-feeding early larvae, including Protozoea I, were used to test the hypothesis that retinoid (Vitamin A) is not required in early decapod crustacean development. In the adult gravid females, retinoids were detected only in the eyes (1.56±0.23 μg g-1 wet mass), whereas there were up to 97 μg g-1 wet mass total carotenoid in digestive gland and epidermis. This was mostly esterified, except in the ovaries, where free astaxanthin predominated (30 μg g-1). No retinoids could be detected in the eggs, the naupliar stages or Protozoea I, but free astaxanthin was metabolised exponentially, falling from 19 μg g-1 in the eggs to 4 μg g-1 in Protozoea I. This suggests that retinoid is not essential in early development and that carotenoid could be taking its place. Also, including retinoids in artificial diets appears to be unnecessary, provided adequate carotenoid is supplied.

Notes: Abstract The fatty acids (FA) in neutral lipid (NL) and polar lipid (PL) of digestive gland, muscle and integument of Penaeus esculentus Haswell were analysed and compared during the moulting cycle and during starvation. The prawns were collected from Moreton Bay, Queensland, Australia, by trawling during 1985–1987, and were fed with a standard semi-purified diet. Compared with a natural diet, the artificial diet had much higher levels of 18:1n-9 and 18:2n-6, but only trace amounts of 20:4n-6, but there was no evidence of dietary imbalance. The fatty acid composition (percentage of total lipid) of the digestive gland changed markedly during the moulting cycle and during starvation, but the small changes observed in both muscle and integument suggested that these tissues maintained their composition under both conditions. When the fatty acids were calculated as absolute amounts, muscle composition, as well as that of the digestive gland, changed significantly. In the digestive gland, saturated FA (SFA), monounsaturated FA (MUFA), diunsaturated FA (DUFA) and polyunsaturated FA (PUFA) all increased up to the middle of the moulting cycle and then declined; with starvation all groups decreased. In muscle, SFA, MUFA and DUFA all increased during the moulting cycle; starvation caused SFA, MUFA and PUFA to decrease, whereas DUFA did not vary. Starvation caused both 18:2n-6 (linoleic) and 18:3n-3 (linolenic) in the digestive gland to reach or almost reach zero. The other essential PUFA, 20:4n-6 (arachidonic), 20:5n-3 (eicosapentaenoic) and 22:6n-3 (docosahexaenoic), decreased during the moulting cycle, but during starvation 20:4n-6 did not decrease as much. In muscle, the levels of 18:2n-6 and 18:3n-3 increased, while 20:4n-6, 20:5n-3 and 22:6n-3 remained approximately constant during the moulting cycle. Starvation reduced 20:5n-3 and 22:6n-3 to about 60%. The data suggest that levels of 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3 are regulated, and that 20:4n-6 can be synthesised from 18:2n-6. There is no clear evidence that 20:5n-3 and 22:6n-3 are essential in P. esculentus, but tissue catabolism of cell membranes during starvation may have provided sufficient amounts for maintenance.

Notes: Abstract Female Penaeus esculentus Haswell were collected by 15 to 20 min duration trawls during 1990. Carotenoids were analysed in the digestive gland, abdominal muscle, the remainder of the body (hereafter called “integument”) and ovary of prawns in Stage 2 through Stage 4 (fully mature) of maturation. The only oxycarotenoids (xanthophylls) identified were astaxanthins or astaxanthin esters; occasionally low levels of β-carotene were detected in the digestive gland. The concentrations of astaxanthin monoesters (AM) and diesters (AD) were highest, with only minor amounts of free astaxanthins (Ast), except in the maturing ovaries, where free astaxanthins predominated (up to 80% of the total carotenoid). Of the total carotenoid, 82 to 94% was in the integument, but at maturity the digestive gland contained 10.7±3.4% and the ovary 5.6±0.9% of the total carotenoid. Only the ovary increased in mass during maturation, reaching up to 5.2% of total prawn mass. During this period, digestive gland concentrations of AM, AD and Ast all increased (tota 20 to 120 μg g-1); levels in the muscle and integument varied little throughout maturation (total ∼0.4 and 100 μg g-1, respectively); ovary AM levels remained low throughout (1.5 to 1.2 μg g-1), AD increased from only 2 to 5 μg g-1, but Ast increased from 2 to 34 μg g-1. Apart from the ovary, AM concentrations were the most variable. In common with other decapod Crustacea, the maturing ovary of P. esculentus contained high levels of carotenoids, indicating that these may have an important role in early development. The natural diet of P. esculentus includes a variety of carotenoids, but except for a little β-carotene, the digestive gland, where absorption occurs, contained astaxanthins, with only an occasional trace of β-carotene. This suggests that the conversion of dietary carotenoids to astaxanthin occurs soon after ingestion.

Notes: Abstract Using the starvation technique, changes in protein and free amino acids were examined in Penaeus esculentus Haswell collected from Moreton Bay, Australia, by trawling in 1985. Prawns of 17.7±0.26 g wet weight were held at 25°C until 2 d after moulting. Groups of seven or eight were then starved fro 5, 10, or 15 d, with appropriate control groups. At the end of each period, ecreted amino acids were collected for 24 h and whole-muscle amino acids and free amino acids (FAA) g-1 in each prawn were analysed. Concentrations of whole-muscle amino acids showed only minor changes with starvation, but concentrations of many of the FAA changed significantly. Total FAA averaged 1 182±45 μmol g-1 dry weight. Individual FAA, in order of abundance, were glycine, arginine, proline, taurine, threonine, hydroxyproline, alanine, glutamic acid, valine, aspartic acid and lysine; the remaining FAA each contributed 〈0.2% of the total. Only taurine and alanine did not show significant changes with starvation. Concentrations of glycine, arginine, hydroxyproline, glutamic and aspartic acid increased, while those of proline, threonine, valine and lysine decreased with starvation, that of proline approaching zero after 15 d starvation. Excreted amino acid-nitrogen represented 〈2% of excreted ammonianitrogen ornithine being the most abundant (35%), followed by leucine (22%) and lysine (17%). The relative abundance of excreted amino acids did not correspond with those of the FAA. It is suggested that, as starvation progresses, the muscle protein is progressively hydrolysed, but with the remaining muscle maintaining its amino acid composition. The liberated amino acids enter the FAA pool and become available for energy production. Proline may have an important role as an energy source, but the ability to synthesise proline may be limited, and thus the artificial food of penaeid prawns may be improved by its addition.