Osmotic and chloride regulation in the hemolymph of the tiger prawn Penaeus monodon during molting in various salinities

The effect of molting on osmotic and chloride concentrations in the blood of the prawn Penaeus monodon Fabricius (20±3 g) at various salinities was investigated. Prawns were obtained from ponds in Iloilo, Philippines, in 1984. They were stocked in salinities of 8, 20, 32 and 44, and their hemolymph was sampled during molt (Time 0), and then 0.125, 0.25, 0.5, 1, 2, 4, 6, 10 and 14 d after molting. Prawns during and immediately after molt tended to conform to the environmental osmolality. Subsequent postmolt (0.5 d) stages displayed more divergence from external salinity. The isosmotic point was higher (940±30 mOsm kg^-1) during molt than during intermolt (663±8 mOsm/kg^-1), suggesting different osmotic requirements in early molt. Hyperregulation of hemolymph chloride below 20 S, as well as isoionic point (301±6 mM), were independent of molting stage. At 20 S and above, newly molted (0 to 0.25 d post-molt) individuals tended to conform to the external chloride concentration while intermolt (0.5 d) post-molt individuals did not. Contribution of hemolymph chloride to hemolymph osmolality was greater during intermolt than during ecdysis, suggesting an important role for other negatively charged ions during molt. When molt occurred in 20 S (the test salinity most similar to the isoionic salinity), there was little or no change in hemolymph osmolality or chloride concentration from 0 to 14 d postmolt. At 8, 32 and 44 S, the change from molt to intermolt values in hemolymph osmotic and chloride concentrations was hyperbolic. Non-linear least-squares regression showed that prawns generally achieved intermolt values within 1 d after molting. Prawns at intermolt regulated hemolymph osmolality (620 to 820 mOsm kg^-1) and chloride concentration (300 to 450 mM) at a much narrower range than during molt (520 to 1 170 mOsm kg^-1 and 250 to 520 mM, respectively). Hemolymph osmolality was a more sensitive indicator of physiological response than hemolymph chloride concentration. Distribution and culture of P. monodon might be limited in low salinities by its ability to maintain a hemolymph osmolality 500 mOsm kg^-1 during molt and 600 mOsm kg^-1 in intermolt, and in high salinities by its capacity to reduce the hemolymph osmolality from values at molt to those in intermolt. Osmotic and chloride concentrations in the blood of P. monodon clearly varied with both molt stage and salinity of the medium. Dependence on external factors, however, gradually declined in older molt stages, suggesting a reduction in integument permeability and greater development of ion absorption/secretion mechanisms as the exoskeleton hardened.SEAFDEC Contribution No. 197     

Body increases during molt and molting cycle of the oriental brown shrimp Penaeus japonicus

Individuals of the oriental brown shrimp Penaeus japonicus Bate, were raised separately (1 shrimp per rearing cage) with surplus food, in almost dark, non-sediment conditions. Results were obtained regarding growth of each part of the body in conjunction with exuviation and molting cycle. Increases in carapace, body length and body weight conformed to the general pattern discribed by Hiatt (1948), with no difference in growth resulting from sex, or inflexion point in juvenile stage. The molting cycle of shrimp weighing 1.5 to 15 g was 6 to 17 days at a water temperature of 20° to 28°C. The molting cycle was prolonged in proportion to the size of the shrimp; shortened as the water temperature increased.     

Variation in lipid classes during the molting cycle of the prawn Penaeus japonicus

The variation in the concentration and fatty acid composition of lipid classes during the molting cycle of the prawn Penaeus japonicus was investigated. The lipid concentration of the whole body reached a maximum at mid-premolt (Stage D₂) and then decreased to low level at late premolt (Stage D_3–4). The accumulation of lipids during the premolt period seemed to be attributable to the increase of both polar and neutral lipids. The increase of neutral lipids at Stage D₂ was derived from not only triglycerides but also free sterols and free fatty acids. Regarding the fatty acid composition of every lipid class, a marked variation occurred mainly at the intermolt (Stage C). In this stage, the polar lipids were rich in monoenoic acids such as 18:1 and poor in polyenoic acids such as 20:53 and 22:63. The triglycerides were rich in polyenoic acids at Stage C, but poor in monoenoic acids such as 16:1 and 18:1. The steryl esters contained large amounts of saturated acids such as 16:0 and 18:0 throughout the molting cycle, however the level of polyenoic acids increased at Stage C.     

Effects of various diets on the nutritional value of Artemia sp. as food for the prawn Penaeus monodon

The nutritional value of Artemia sp. as food for marine fish and crustacean larvae has been linked to the level of its polyunsaturated fatty acid (PUFA) content. Experiments in August 1984 were conducted to determine the effects of various artificial diets and algae on fatty acid composition of PUFA-deficient Artemia sp. (Utah GSL strain) and their resulting value as food for postlarvae of the prawn Penaeus monodon (Fabricius). Nauplii of the brine shrimp were grown on extracts of corn, copra, soybean and rice bran containing precursors (C18) to long-chain PUFA and also on algal species containing different levels of long-chain PUFA (C20). The nauplii were then used as food for P. monodon postlarvae. The results revealed that absence of C20 polyunsaturates from the feeds and their presence in the algae were reflected in the polyunsaturated fatty acid content of the tissues of Artemia sp. When fed with brine shrimp fed on algae, P. monodon displayed better postlarval survival and significantly higher growth; related to the content of polyunsaturated fatty acids in Artemia sp. A practical feeding approach in prawn hatcheries would be to grow Artemia sp. on a cheap diet such as rice bran, and then to enhance its nutritional value with a diet high in PUFA prior to harvesting, in order to improve hatchery production.     

Fatty acid composition of organs and tissues of the tiger prawn Penaeus esculentus during the moulting cycle and during starvation

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.     

Population structure of the giant tiger prawn Penaeus monodon in Australian waters, determined using microsatellite markers

 We describe three highly polymorphic microsatellite loci which have been isolated from the giant tiger prawn Penaeus monodon. The number of alleles present among 312 samples at the loci Pmo9, Pmo25 and Pmo27 were 84, 34 and 35, respectively, with heterozygosities all >90%. Analyses of the distribution of length variation at three microsatellite loci among five Australian P. monodon populations revealed strong differentiation between populations from the west and those from the northern and eastern coasts. Tests for population differentiation (F st) values and an analogous measure for microsatellite loci (R st) all demonstrated that Western Australian P. monodon are a separate genetic stock which exhibits reduced genetic variation relative to the other populations. Reduced variability is consistent with a recent population bottleneck or colonization by a small founding population from the east when sea links between Indonesia, New Guinea and Australia were re-established following the last ice age. The results of this study are in agreement with previous surveys of P. monodon conducted with allozymes and mtDNA. Received: 18 December 1998 / Accepted: 27 August 1999     

Carotenoids in the tiger prawn Penaeus esculentus during ovarian maturation

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 -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 g g^-1); levels in the muscle and integument varied little throughout maturation (total 0.4 and 100 g g^-1, respectively); ovary AM levels remained low throughout (1.5 to 1.2 g g^-1), AD increased from only 2 to 5 g g^-1, but Ast increased from 2 to 34 g 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 -carotene, the digestive gland, where absorption occurs, contained astaxanthins, with only an occasional trace of -carotene. This suggests that the conversion of dietary carotenoids to astaxanthin occurs soon after ingestion.     

Influence of salinity and temperature on the oxygen consumption in young juveniles of the Indian prawn Penaeus indicus

Routine oxygen consumption of very young juveniles (0.1 g) of Penaeus indicus H. Milne Edwards was significantly influenced by ambient temperature and weight of the animal, but not by ambient salinity, when tested at salinities (7, 21, and 35) to which they had been long-term (over 10 days) acclimated. Standard oxygen consumption of young juvenile prawns (1 to 3 g), subjected to step-wise changes in ambient salinity, from sea water to low salinity waters (2 to 6), and measured after short-term (24 h) salinity acclimation at each step, was lowest at salinities where prawns such as those tested occur naturally (10 to 15). The metabolic rates do not appear to have a direct relation to the osmotic gradient, even when the influence of interfering activity is eliminated. It appears that factors other than osmotic gradient will have to be sought in order to explain the metabolic patterns of P. indicus in relation to salinity.     

Effect of salinity on oxygen consumption in postlarvae of the penaeid shrimps Penaeus monodon and P. stylirostris without and with acclimation

The effects of salinity on oxygen consumption was investigated in cultured Penaeus monodon Fabricius and P. stylirostris Stimpson postlarvae (32 and 35 d after metamorphosis, respectively). In both species, no difference appeared between non-acclimated and fully-acclimated individuals. The metabolic rate was unaffected by salinity variations, but P. stylirostris displayed a tendency (non-significant at P<0.05) to increase respiration at lower salinities. These observations are discussed in reference to the ecology of the young prawns.     

Population structure of the black tiger prawn, Penaeus monodon, among western Indian Ocean and western Pacific populations

We examined the population structure of the black tiger prawn, Penaeus monodon Fabricius, 1798, in the Indo-West Pacific by analyzing the geographic distribution of elongation factor 1-alpha intron sequences from specimens collected during the winter and spring of 1997. Both the molecular phylogeny of alleles and F-statistics indicated very strong differentiation between populations from the western Indian Ocean and western Pacific. This pattern is concordant with other recent studies of marine species in this region, implying that the Indo-Australian Archipelago represents a biogeographic break between populations in the Indo-West Pacific. F _ST-values among populations in the western Indian Ocean also indicate structure within this region, whereas no structure was found among western Pacific populations. Nucleotide diversity was significantly lower in the western Indian Ocean populations than in the western Pacific, implying that the populations have regional differences in demographic history. Received: 16 November 1998 / Accepted: 26 May 1999     

Responses of hemolymph osmolality and tissue water of Penaeus chinensis Osbeck juveniles subjected to sudden change in salinity

Hemolymph osmolality and tissue water of laboratory-reared Penaeus chinensis Osbeck juveniles (0.83 to 1.86 g) were investigated, after they had been transferred individually from 10, 20, 30 and 40 ppt to 10, 20, 30 and 40 ppt for 0.25, 0.5, 1, 2, 5 and 10 d, respectively. Hemolymph osmolality and tissue water of shrimp were stablilized within 5 d after they had been subjected to a sudden change in salinity from each salinity level. Hemolymph osmolality had a positively linear relationship with medium osmolality. Tissue water decreased with increased medium osmolality, and decreased with increased hemolymph osmolality. The mean (SD) isosmotic point was 703 (8) mOsm kg^–1 which is equivalent to 24.2 (1.0) ppt. P. chinensis juveniles exhibited hyperosmotic regulation in salinities below isosmotic value, and hypoosmotic regulation in those above. The shrimp originally adapted to high salinity levels (30 and 40 ppt) showed less fluctuation of tissue water than those adapted to low salinity levels (10 and 20 ppt) when they were subjected to a sudden change in salinity.     

Effects of body size and temperature on the metabolic rate of Penaeus monodon

Laboratory measurements of oxygen consumption were made on Penaeus monodon (Fabricius) from protozoea to adult stage at temperatures between 15° and 35°C. The logarithmic relationship between weight-specific respiration rate (WRt) and temperature (T) for two size groups, Protozoea 1 (PZ1) to Postlarva 1 (PL1) and PL to adult, are given as; WRt=10^{0.431+0.0146 (T)} (ml O₂ g^-1 h^-1) and WRt=10^{-0.948+0.0338 (T)} (ml O₂ g^-1 h^-1), respectively. Additionally, equations relating metabolic rate, temperature and size for the two size groups are; PZ1-PL1: log M=0.431+0.0146T+(1.25 (log TL)+0.579), and PL1-adult: log M=-0.948+0.0338T+(2.60(log CL)-0.683), where M=oxygen consumption in ml O₂ individual ^-1h^-1, T=temperature in °C, TL=total length in cm, and CL=carapace length in cm. Activation energies of 6 186.75 J for PZ1-PL1 and 14 066.62 J for PL-adults point to different metabolic pathways or to differences in the ratio between the metabolic pathways used.     

Changes in protein-bound and free amino acids in the muscle of the tiger prawn Penaeus esculentus during starvation

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 mol 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.     

Effects of salinity fluctuation on the hemolymph composition of the blue crab Callinectes sapidus

The hemolymph of the blue crab Callinectes sapidus was hyperosmotic during 20-10-20 S and 30-10-30 S diurnal cycles. The hemolymph became isosmotic at 26 S and hyposmotic at 28 S in the 10-30-10 S diurnal cycle. Hemolymph Na⁺ was hyperionic to seawater throughout all cycles. Hemolymph Cl^- was hyperionic below 24 S and either isionic or hypoionic from 24 to 30 S. Hemolymph K⁺ concentrations were hyperionic below 26 S and either isionic or hypoionic from 26 to 30 S. Hemolymph Mg⁺⁺ values were hypoionic over the experimental salinity range (10 to 30). Hemolymph ninhydrin-positive substances (NPS) levels were directly related to ambient salinity.     

Structure and function of the chelate pereiopods of the banana prawn Penaeus merguiensis

Penaeus merguiensis de Man systematically searches the substratum and removes small particles of food from it using the small chelae of the first three pairs of pereiopods. One or more chelae are used to transfer food particles to the mouth. The propus and dactylus of these limbs bear numerous setae arranged in discrete groups along the length of the segments. The tip of each seta is elaborately sculptured and has a large sub-terminal pore. It is probable that these are chemosensory organs responsible for discriminating edible from non-edible material. Proximal to almost every setal group is a much-branched seta (rarely two) which probably measures the depth to which the chela has been inserted into the substratum. At the articulation of the propus and carpus of the first pereiopod are three groups of differently sculptured setae, whose function is to clean the other chelate pereiopods. An elaborate system of pegs and a ridge on the apposable fringes of the chelae might be a mechano-receptive device with a particle size discriminating function. The ridges merge into opposable terminal denticulate pads whose function is to grasp food and other particles, which may be quite small (10 to 20 m upwards).     

Structure and function of the third maxillipeds of the banana prawn Penaeus merguiensis

The functions of the third maxilliped of Penaeus merguiensis de Man are described. It has a 6-segmented endopodite which is used in feeding, grooming and possible pheromone reception. In feeding, large pieces of food are gripped by the stout spines on inner borders of the ischia which hold them to the mouth so that particles can be torn off and ingested. All the distal segments have long setae used in grooming chelate pereiopods and the antennules. In mature males there is a tuft of about 300 slender setae at the end of the propus, but its function remains obscure.     

Impact of mineral deposition on shrimp, Penaeus monodon in a high alkaline water

This study compares water quality parameters, shrimp growth and mortality rates, and biomass at harvest in two ponds of equal size, seeded with the same density (7 m2) of White Spot Syndrome Virus (WSSV) and Monodon Baculo Virus (MBV) negative post-larvae (PL)-20 of shrimp, Penaeus monodon in the Vellar estuary of South India. The primary difference between the ponds was the water source; one was filled from the estuary and the second with water from bore wells with high alkalinity. Temperature in both ponds was similar and reached 320C after 185 days of culture. Dissolved oxygen (DO) levels were within the acceptable range although levels in the alkaline pond were near the lower limit for the last 90 days before harvest. Salinity levels were similar in both ponds, above optimal levels, and increased over the 185 days. Alkalinity in the estuarine water was typically <50 ppm and again 200-320 ppm in the alkaline pond. In the alkaline pond, beginning on the 75th day mineral deposits was observed covering all parts of the shrimp including the eye and the inner gill chambers, and by harvest, 42% of the shrimp showed this coating. Elemental analysis identified the major constituents as calcium, phosphorus and manganese. Survival rates in the estuarine-water-fed pond was 92% with a total pond biomass at harvest of 1.65 tons ha-1 compared to survival of 79% in the alkaline pond and a biomass at harvest of 1.020 tons ha-1. When well water must be used, its alkalinity should be monitored and diluted with water from other sources.     

Effects of salinity and calcium concentration on arm regeneration byOphiothrix angulata (Echinodermata: Ophiuroidea)

At 33 salinity a tissue stump formed 2 to 3 d after autotomy and developing ossicles were present by the fourth day inOphiothrix angulata (Say). Regeneration proceeded rapidly from the sixth day until the thirteenth day, when the rate decreased greatly. The length of the regenerated arm and the number of ossicles formed did not vary over a salinity range of 28 to 38 S, but were significantly less at 23 S. The number of ossicles regenerated increased linearly (y=1.9 x-7.7;r=0.9089) with the calcium concentrations ranging from 3.8 to 9.5 mM. No ossicle formation occurred at 3.8 mM calcium concentration. Rate of net uptake of calcium-45 into the ossicles of intact individuals in salinities of 28 and 33 was significantly greater than that in 23 and 38 S. However, net uptake rate of calcium into the soft tissues of the arms was significantly higher at 18S than at the lower two salinities.     

Effects of endogenous and some exogenous factors on the activity of the juvenile banana prawn Penaeus merguiensis

In conditions of alternating light and dark, juvenile Penaeus merguiensis de Man are more active during the dark phase. The rhythm persists in continuous dim red light, but not in continuous bright white light. The period of circadian rhythm shown in continuous dim red light varies between individuals from 22.75 to 26.0 h, with a mean of 23.8 h. The response of prawns to an artificially produced tidal situation is mediated by the presence or absence of water flow. No endogenous component of this tidal rhythm was demonstrated. There is an irregular short-term rhythm (period 2 to 3 h). It is suggested that this allows starved prawns to conserve energy.     

External synchronizers of tidal activity rhythms in the prawns Penaeus indicus and P. monodon

Phasing of persistent circatidal rhythmicity to an artificial tidal cycle was assessed in the prawns Penaeus indicus Milne Edwards and P. monodon (Fabricius) collected from the Vellar estuary, South India, in the period between June and December 1984. Simulated 6 h cycles of 20 and 30 S, and 6 h cycles of 20° and 30°C induced a persistent tidal rhythmicity after 20 cycles. The imposed 6 h cycles of 25 and 30 S, and 25° and 30°C induced tidal rhythms after 30 cycles. In both cases, re-established tidal activity rhythms were evident for at least 48 h — higher activity occurring during the higher salinity and lower temperature phases of the simulated tidal cycles. Artificial tidal cycles of still water and running water synchronized the tidal rhythm after 20 cycles. Combined 30 S, 20°C, for 6 h and 20 S and 30°C for 6 h established a persistent tidal rhythm after 10 cycles, whereas wave action had no influence on tidal synchronization. The influence of possible interactions of tidal rhythms and in situ tidal variables on circatidal activity is discussed.