Changes in lipid class and fatty acid contents in the ovary and midgut gland of the female fiddler crab Uca tangeri (Decapoda,Ocypodiadae) during maturation

Changes in biochemical composition, lipid class and fatty acid contents were studied in the ovaries and midgut glands of the fiddler crabs Uca tangeri Eydoux during maturation. Wild females were caught during spring and early summer of 1992 in the Bay of Cádiz (southwest Spain), near the mouth of the San Pedro river. Protein and total lipid contents in the ovaries increased significantly from Stages III to IV, at the expense of total carbohydrate, which showed a large decrease during the same period. In the midgut gland, the protein content did not present any significant variation, whereas total lipids and total carbohydrates presented opposite up and down trends during maturation. In the ovary, total polar lipids increased significantly during the final phase of maturation (Stages III to IV), mainly due to the significant contribution of the phosphatidylcholine and phosphatidylethanolamine fractions. In contrast, total neutral lipids showed an upward trend throughout the whole maturation period, mainly due to significant increases of the triacylglycerol fraction. In the midgut gland, total polar lipids (mainly phosphatidylcholine) and total neutral lipids (mainly triacylglycerol) presented significant decreases from Stages II to III, the phase which preceded major increases in both polar and neutral lipids in the ovaries. Cholesterol content did not vary during maturation in either organ, in the ovary or midgut gland. Major fatty acids in the ovaries [16:0, 16:1 (n-7), 18:1 (n-9), 18:1 (n-7), 18:2 (n-6), 18:3 (n-3), 20:4 (n-6), 20:5 (n-3) and 22:6 (n-3)] did, however, accumulate significantly at later stages of maturation. It is noteworthy that arachidonic acid [20:4 (n-6)] content remained constant during all stages of maturation but decreased significantly in total polar lipids in the later phases of maturation. In contrast, eicosapentaenoic acid [20:5 (n-3)] increased significantly in all lipid fractions in the later stages, and docosahexaenoic acid [22:6 (n-3)] remained constant in the polar lipids and increased during later stages in the triacylglycerol fraction. Major fatty acids in the midgut gland lipids showed significant decreases from Stages II to III, just before the final period of maturation.     

Variation in the lipid content of wild-caught females of the marine shrimpPenaeus kerathurus during sexual maturation

Changes in total lipids, lipid classes and their fatty acid contents were studied in the ovaries and midgut glands ofPenaeus kerathurus Forskäl females during sexual maturation. The shrimp were captured in the Gulf of Cádiz (southwest Spain) in 1990. The lipid content and fatty acids, in relative terms, increased during ovarian development. The greatest changes occurred between Maturation Stages III and IV. Ovarian lipids were dominated by polar classes, whereas in the midgut gland the major classes were triacylglycerols and sterol esters. The amounts of major fatty acids in ovaries (16:0, 16:1n-7, 18:1n-9, 18:1n-7, 20:5n-3 and 22:6n-3) increased with increasing maturity, but declined slightly between Stages III and IV. The total polar lipid content of the midgut was 5.7% (by dry weight) and its fatty acid composition remained constant during the whole study period. Total lipid content of the midgut gland showed an upward trend during sexual maturation, except between Stages II and III, when a slight decrease was observed. Predominant fatty acids in the midgut gland (16:1n-7, 20:5n-3 and 22:6n-3) displayed a noteworthy decline between Stages II and III, corresponding with the marked increase in total lipid fatty acid content in the ovaries during the same period.     

In situ variation in heart rate of the shore crab Carcinus maenas in relation to environmental factors and physiological condition

Heart rate of Carcinus maenas was recorded continuously for two days in situ, together with water temperature, salinity, depth and light intensity. Of each sex, 25 crabs were used and assigned to groups on the basis of size and colour (the carapace changes colour with prolongation of the intermoult phase). Wet weight:dry weight ratios of midgut gland, muscle, gonads and whole body were examined together with midgut gland lipid content and haemolymph protein concentration. Heart rates and wet wt:dry wt ratios for all the tissues examined were higher in early intermoult than in late intermoult stages of the adult C. maenas (P<0.05). Heart rate, lipid content and haemolymph protein concentrations were higher and wet wt:dry wt ratios of midgut gland, gonads and whole body were lower in juvenile crabs than in adults (P<0.05). The relationship between heart rate and wet wt:dry wt of whole body differed among C. maenas in early and late intermoutl. Heart rate was positively correlated with midgut gland lipid content (a Darwinian fitness parameter) in crabs that were in late intermoult. Physico-chemical environmental parameters, sex and colour accounted for 12% of the variation in heart rate of C. maenas recorded in situ. Temperature and prolonged intermoult were the most important factors influencing heart rate. The findings are discussed in relation to laboratory studies of cardiac activity and observed behaviour of C. maenas in the field.     

Factors determining the hatching success of Antarctic krill <Emphasis Type="Italic">Euphausia superba</Emphasis> embryo: lipid and fatty acid composition

The present study addresses the effect of maternal diet on hatching success and condition of embryos and larvae of Antarctic krill Euphausia superba. Lipid and fatty acid content and composition were determined in field and laboratory samples. Developmental stages analyzed in embryos included: multiple-cell, gastrula, and limb-bud stages. Larval stages analyzed included: nauplius I, nauplius II, and metanauplius. Laboratory-reared embryos were spawned by gravid females incubated under three feeding groups: (1) phytoplankton mixture, (2) phytoplankton mixture and minced clam, and (3) phytoplankton mixture, minced clam, and commercial larval food. Hatching success was highest in group 3 (100%), lowest in group 1 (0%), and highly variable in field samples (0–48%). Lipid decreased slightly in embryos during embryonic development, while large decreases in lipid were found during nauplius development. High levels of 18:2(n-6), 20:4(n-6), and 22:6(n-3) observed with group 3 samples coincided with high hatching success in krill embryos. The ratio of 22:6(n-3)/20:5(n-3) also correlated to hatching success of embryos. The fatty acid profile of embryos in group 3 was similar to that of the field-collected embryos, reflecting the contribution of the commercial larval food in the maternal diet. In our study, the maternal diet was found to influence the fatty acid composition of embryos and in turn affects the hatching success of krill. Specific polyunsaturated fatty acids appeared to play important roles in embryogenesis in krill.     

Mechanisms of copper toxicity in the shore crab,Carcinus maenas

The influence of copper(II)chloride on selected key enzymes and metabolites in intermediary metabolism and the energy charge potential of the hyperregulating shore crab, Carcinus maenas (L.) were investigated. Crabs, collected in Odense Fjord, Denmark between September 1989 and May 1990, were exposed to 10 ppm copper(II)chloride, in 10 ppt salinity seawater at 15°C for up to 1 wk. Hexokinase activity was 77% lower in the posterior gills and 60% lower in the midgut gland than in control crabs. Phosphofructokinase activity was reduced in the midgut gland by 82% but was unaffected in the gills. Pyruvate kinase activity was undetectable in the gills of exposed crabs and reduced by 47% in the midgut gland. Citrate synthase activity was depressed by 30% in the posterior gills and unaffected in other tissues. Copper exposure had no apparent effect on tissue cytochrome c oxidase activity. Chela muscle was the only tissue in which copper exposure did not bring about reductions in the enzyme activities tested. As a result of 1 wk of copper exposure the average lactate level increased by 3.5-fold in haemolymph, 9.3-fold in posterior gills and 6.5-fold in midgut gland. The average glucose level was raised by a factor of 6 in the haemolymph, 3.4 in the posterior gills and 1.2 in the midgut gland. Energy charge potential was at a very high level (ca. 0.9) and was unaffected by copper exposure.     

Changes in lipid content,lipid class composition and fatty acid composition of developing eggs and unfed larvae of cod (Gadus morhua)

Changes in the lipid class and fatty acid composition of developing eggs and unfed larvae of cod (Gadus morhua L.) were studied with the objective of determining probable requirements of cod larvae for dietary lipid. The eggs were collected on 24 March 1985 from holding tanks containing cod which had been caught off the northwest coast of Scotland. Phosphatidylcholine (PC) was the only lipid class to decline in absolute terms during embryogenesis. Catabolism of neutral lipid was initiated during the first week after hatching, and the rate of neutral lipid utilisation increased after the larvae had completely absorbed their yolk sacs. The quantity of triacylglycerol (TAG) remained constant during embryogenesis, but the percentage of 22:6(n-3) int TAG increased substantially during this period. It was calculated that ca. 33% of the 22:6(m-3) released during the process of PC catabolism was incorporated into TAG and sterol ester. The results suggest that PC, replete in appropriate essential fatty acids, should represent a major proportion of the lipid in artificial diets for fish and crustacean larvae.     

Interindividual variability in metal status in the shore crab<Emphasis Type="Italic"> Carcinus maenas</Emphasis>: the role of physiological condition and genetic variation

The relations between genetic variation, physiological condition, size and metal status in the shore crab Carcinus maenas (L.) were investigated. Shore crabs were collected at three sites around the island of Funen, Denmark, and carapace width, colour, moulting stage and major ions and protein in the haemolymph were determined. Concentrations of water, cadmium, copper and zinc in midgut glands, muscles and gills were measured, and allozyme variation at 15 loci was studied. Generally, tissue water content and copper, zinc and cadmium concentrations decreased from the moulting stages C₃ over C₄ to D crabs, whereas the opposite was true for copper and zinc concentrations in gills and muscle. However, the water content of the tissues increased with the size of the crabs. Since tissue water contents changed consistently over moulting stages, conclusions on changes in metal concentrations sometimes diverged when expressed on a dry versus wet weight basis. Regarding allozymes, esterase, malate dehydrogenase and phosphoglucomutase were polymorphic. The crabs showed a low level of genetic variability for both polymorphism (0.14) and for heterozygosity (0.07) at the three sites. Allelic frequencies showed very low levels of differentiation among the samples, no evidence of inbreeding and no population subdivision. Multilocus heterozygosity was positively correlated with size. Concentrations of copper and zinc in midgut gland, muscle tissue and gill were not correlated with genotype, whereas a weak correlation between phosphoglucomutase genotypes and concentrations of cadmium in the midgut gland was found.Communicated by M. Kühl, Helsingør     

Studies on the uptake of cadmium by the crab Carcinus maenas in the laboratory. I. Accumulation from seawater and a food source

The crab Carcinus maenas (L.) was exposed to radioactively labelled cadmium dissolved in seawater at concentrations of 0.1, 1 and 10 ppm, the latter concentration being toxic to the crabs (50% mortality after 12.3 days). Net accumulation of cadmium from solution was proportional to the level and time period of cadmium exposure. Total absorbed cadmium levels reached 0.0043 and 0.0412 mg Cd g^-1 dry weight after 40 days exposure to 0.1 and 1 ppm Cd, respectively, and 0.1115 mg Cd g^-1 dry weight after 12.3 days average exposure to 10 ppm Cd. The highest tissue concentration was found in the midgut gland, reaching 0.786 mg Cd g^-1 dry weight after 12.3 days average exposure to 10 ppm Cd. The midgut gland only contained about 10% of the total cadmium absorbed from solution, while the exoskeleton contained the bulk of obsorbed cadmium (59 to 80%) probably passively adsorbed onto the surface. When cadmium was absorbed by the crabs from a food source, the midgut gland contained 16.9% of the total absorbed cadmium whereas the exoskeleton now contained only 22.2%. Ten percent of the cadmium available in the food source (Artemia salina) was accumulated by the crabs. When placed in cadmium-free seawater, crabs that had accumulated cadmium from solution lost 69% of the absorbed cadmium in 10 days, mostly from the exoskeleton which lost 78% of its original absorbed cadmium concentration.     

Arsenic accumulation in the shore crab Carcinus maenas: the influence of nutritional state,sex and exposure concentration

The accumulation of arsenate from seawater by the shore crab Carcinus maenas L. (collected from Odense Fjord, Denmark in 1991 and from Restronguet Creek, UK in 1991) was investigated in a series of laboratory experiments. A field study was also carried out to determine the effects of raised environmental arsenic concentrations on intra-organismal distribution and tissue concentrations. Studies on the influence of nutritional state and sex on accumulation of As(5) from seawater indicated that most of the arsenic taken up from seawater in laboratory experiments was retained in the gills and the midgut gland. Arsenic accumulation exhibited sex-dependent differences which were also evident in correlation analyses carried out between total lipid contents and total arsenic contents of midgut glands of individual crabs. Arsenic concentrations in the gonads of both sexes were strongly influenced by the nutritional state of the crabs. Elevated arsenic concentrations in seawater and food at an arsenic polluted site (Restronguet Creek) significantly influenced arsenic concentrations and distribution among the tissues of C. maenas. Arsenic concentrations and distribution patterns differed markedly from those crabs from an unpolluted site in Odense Fjord. The gills of the crabs from Restronguet Creek contained extremely high arsenic concentrations ranging from 179 to 483 g As g^-1 dry wt. These values were even higher than those measured in the gills of Odense crabs that had been exposed to 3 mgl^-1 As(5) for 2 wk in the laboratory. Arsenic concentrations in the exoskeleton of Odense Fjord crabs were 15 times lower than those measured in exoskeletons of Restronguet Creek crabs. Approximately 69% of the total body burden of arsenic was located in muscle tissue of crabs from Odense Fjord, whereas the major pool of arsenic (46%) in Restronguet Creek crabs was located in the exoskeleton.     

Reproductive biology of shovelnose guitarfish <Emphasis Type="Italic">Rhinobatos productus</Emphasis> from the eastern Gulf of California México

Guitarfishes are a primary component of artisanal elasmobranch fisheries and are commonly taken as trawl fishery bycatch throughout the Gulf of California. However, little is known of the life history of this species. To address this lack of critical biological information, the reproductive biology of Rhinobatos productus was investigated in the eastern Gulf of California. Development of claspers and testes indicated that males reach maturity at 53 cm total length (TL). Measurements of oviducal gland, largest ovum diameter, and uterus width indicated that females >57 cm TL are mature. This species possesses two functional ovaries: the ovarian cycle and gestation run concurrently. Histological analysis of oviducal glands did not provide evidence of sperm storage, but females carrying uterine capsules were observed over an extended period, suggesting the possibility of diapause in the early embryonic development. Following 4–5 months of embryonic growth, pups were typically born from late June to October after a gestation period of approximately 11–12 months. Width of yolk sac was inversely related to embryo length. The rate of reduction of yolk sac width suggests that embryos depend on the sac until birth. Mean fecundity was estimated to be 5 (range 1–10, s.d. = 2.24) with a 1:1 sex ratio. Average size at birth was 175 mm TL. Seasonally, gravid females enter shallow waters for parturition, becoming extremely vulnerable to gill nets used in the artisanal ray fishery.     

Trace metal concentrations and contents in the tissues of the shore crab<Emphasis Type="Italic"> Carcinus maenas</Emphasis>: effects of size and tissue hydration

Water content in the tissues, tissue proportions, and content and concentrations of copper, zinc, iron, cadmium, manganese, and cobalt were determined in male shore crabs Carcinus maenas of different sizes. Average wet to dry weight ratios in midgut gland, gills, muscle, exoskeleton, stomach, heart, testes, hypodermis, and haemolymph were 5.3, 9.5, 4.1, 1.5, 4.8, 6.1, 3.8, 4.7, and 13, respectively. The water content in the soft tissues generally increased with increasing size of the crabs (except in testes). Dry midgut gland, gills, muscle, exoskeleton, stomach, heart, testes, hypodermis, and haemolymph constituted 4, 0.9, 20, 68, 0.5, 0.12, 0.5, 0.7 and 5.3%, respectively, of the body dry weight. Dry midgut gland, gills, stomach, hypodermis, and haemolymph constituted a larger proportion of the body weight in small than in large crabs, whereas the opposite was the case for exoskeleton. Protein concentrations in the haemolymph decreased with size whereas haemolymph volume increased. Seventy percent of the copper body burden was located in the haemolymph. The majority of zinc (61%) occurred in muscle, whereas the midgut gland was the major site of cadmium storage (89%). The exoskeleton contained the major part of the body burden of iron (71%) and manganese (95%). Most of the cobalt body burden was located in the exoskeleton (45%) and haemolymph (44%). As tissue hydration and tissue proportions changed with size, tissue metal concentrations did not necessarily vary identically with size when expressed on a dry weight compared to a wet weight basis. Tissue potassium concentrations were negatively correlated with tissue water contents, indicating that water in tissues with a high wet to dry weight ratio was of extracellular origin. However, comparison of tissue and haemolymph copper levels showed that this extracellular water could not be pure haemolymph as tissue copper concentrations would have been higher than the values actually found.     

Interactions between temperature and salinity during brooding on subsequent zoeal development of the mud crab Rhithropanopeus harrisii

Female mud crabs, Rhithropanopeus harrisii, carrying newly extruded eggs, were collected from the Petaluma River (San Francisco Bay Estuarine System, California, USA) in summer 1985, and exposed to factorial combinations of temperature (20°, 25° or 30°C) and salinity (2, 5, 15, 25, or 32%.). Upon hatching, dry weights of 12 to 15 h-old zoeae were determined. Subgroups of the remaining zoeae were transferred from hatching salinities to the salinities listed above and raised until metamorphosis to megalopa. Low salinities reduced zoeal dry weights by as much as 25%. Temperature played a secondary role in reduction of hatching weight of zoeae. Survival of larvae through zoeal development was best when hatching and rearing salinities were the same; in this case, overall survival increased with temperature. Both duration of zoeal development and megalopal dry weights were strongly influenced by temperature and rearing salinity, with only a small contribution from hatching salinity. The influence of hatching salinity was most obvious at extremes of the range tested. These studies indicate that physical conditions during embryogenesis profoundly influence subsequent larval development. Interpretation of experimental approaches to study ecophysiological adaptations of larval stages should not neglect the role of physical conditions during embryogenesis.     

Cadmium accumulation in the female shore crab Carcinus maenas during the moult cycle and ovarian maturation

 The influence of moulting and ovarian maturation on cadmium accumulation in the tissues of female shore crabs Carcinus maenas exposed to 1 mg Cd l⁻¹ in the water was investigated. Cadmium accumulation in all tissues examined was markedly increased in crabs in the postmoult stages (A and B) compared to crabs in all other moult stages. During the moult cycle, average cadmium accumulation in the midgut gland ranged from 29 μg Cd g⁻¹ dw at premoult stage (D₂) to 589 μg Cd g⁻¹ dw at postmoult stage (A). Average cadmium concentrations in the haemolymph ranged from 0.56 μg Cd ml⁻¹ at intermoult stage (C₄) to 4.6 μg Cd ml⁻¹ at postmoult stage (A), while the gills accumulated from 103 μg Cd g⁻¹ dw in intermoult stage (C₃) to 352 μg Cd g⁻¹ dw in postmoult stage (A). Cadmium concentration in gills and haemolymph was also significantly higher in crabs in late premoult stage (D₃) compared to C₄-crabs, while midgut gland cadmium concentration remained elevated in C₁- and C₃- intermoult stages relative to C₄. During ovarian maturation the cadmium accumulation in midgut gland, gills, ovaries and haemolymph decreased. Average cadmium concentration in the midgut gland decreased from 63 μg g⁻¹ dw in ovarian Stage I to 19 μg g⁻¹ dw in ovarian Stage VI. The same pattern was observed for gills, haemolymph and ovaries. The present study demonstrates that cadmium accumulation in the female shore crab strongly depends on the physiological status of the animal. A possible association between physiological calcium requirements and cadmium accumulation during moulting is discussed. Received: 20 January 2000 / Accepted: 20 July 2000     

The importance of free amino acids to the energy metabolism of eggs and larvae of turbot (Scophthalmus maximus)

A study was undertaken to establish the role of free amino acids (FAA) in aerobic energy dissipation in embryos of turbot (Scophthalmus maximus) which contain an oil globule in the egg. Laboratory-reared developing eggs and larvae (15°C, 34 salinity) were measured for oxygen uptake, ammonia excretion, contents of FAA, protein, and ammonium, and volumes of yolksac and oil globule. Newly spawned eggs from different batches contained 55 to 90 nmol egg^–1 of FAA. Resorption of FAA occurred in parallel with the consumption of yolk. Resorption of the oil globule, however, occurred predominantly after hatching and mainly after yolk resorption. The combined data suggest that approximately 70% of the FAA are utilized as an energy substrate, while the rest are polymerized into body proteins. FAA become a significant energy substrate in the early egg stage and account for 100% of the aerobic energy dissipation 2 d after Fertilization then decrease to ca. 60% at the time of hatching. Lipids derived from the oil globule seem to be the main fuel after hatching and account for ca. 90% of the energy dissipation at the onset of first-feeding. Thus, the energetics of fish embryos which contain an oil globule seems to be different from those that depend exclusively on the nutritional reserves of the yolk.     

Egg survival,embryonic development,and larval characteristics of northern shrimp (<Emphasis Type="Italic">Pandalus borealis</Emphasis>) females subject to different temperature and feeding conditions

Laboratory experiments on ovigerous females of northern shrimp (Pandalus borealis) were used to assess the effects of temperature and food ration on female condition during incubation and examine how combined effects of temperature and female condition influenced egg survival, embryonic development, and larval characteristics. Ovigerous females were maintained at 2°C, 5°C, and 8°C and fed on a low (three times/week; 2–2.7% W/W) or high ration (five times/week at satiation). The increase in temperature accelerated the developmental time of the eggs but their survival at 8°C was reduced. Conversion efficiency of yolk reserves in developing embryos was significantly reduced at elevated temperatures and larvae hatching at 2°C and 5°C were significantly larger and heavier than those hatching at 8°C. The experimental design did not result in any effect of food ration on the energetic condition of females or on egg characteristics and their biochemical composition. However, lower energy reserves were observed for females held at 8°C.     

The functional morphology and development of the alimentary tract of larval and juvenile barnacles (Cirripedia: Thoracica)

The alimentary tract of the nauplius larva of Balanus spp. consists of cuticle-lined foregut and hindgut, with intervening endodermal midgut constricted into anterior and posterior regions. The anterior midgut cells in the region of the constriction (constriction cells) secrete proteins (probably digestive enzymes). The remaining anterior midgut cells, often containing lipid droplets, form the absorptive region of the tract. Glycoprotein globules and lipid droplets within anterior midgut cells are the remants of the yolk in a pre-hatched larva, this yolk additionally supporting the larva through the non-feeding first nauplius stage. Nauplius Stages II to VI are actively feeding planktonic stages which increase in size and build up lipid reserves. These accumulated reserves support the non-feeding cyprid, first through its planktonic life and then through settlement and subsequent metamorphosis to the juvenile barnacle. Juvenile barnacles start to feed between 2 and 5 days after metamorphosis.     

纳米银和银离子对斑马鱼胚胎早期生长发育的影响及作用机制

为探究纳米银对水生生物的毒性作用,选取斑马鱼胚胎为受试生物,考察了纳米银对斑马鱼胚胎早期生长发育的影响,同时比较了纳米银与银离子对斑马鱼胚胎的毒性作用和机理。实验将受精后4小时(4 hpf)的斑马鱼胚胎分别暴露于不同浓度的纳米银和银离子溶液中至96 hpf,观察并记录了胚胎的死亡、孵化和畸形等指标。应用吖啶橙(AO)染色实验研究了胚胎暴露之后的细胞凋亡情况,并且应用荧光定量PCR技术分析了相关基因的表达水平。研究结果表明,随着暴露浓度的增加,纳米银和银离子均能导致斑马鱼胚胎的死亡率增加和孵化率降低,并且引起孵化延迟。纳米银和银离子的96 h半数致死浓度(96 h-LC50)分别为11.75 mg·L-1和0.054 mg·L-1。银离子毒性远大于纳米银毒性。暴露的斑马鱼胚胎均表现出体长变短和卵黄囊肿大的畸形。AO染色结果表明,纳米银和银离子处理组胚胎的躯干和卵黄囊部位存在细胞凋亡信号。基因表达分析结果显示,1.93 mg·L-1纳米银显著提高了斑马鱼胚胎caspase9的表达(P0.05),而0.006 mg·L-1的银离子就能显著上调COX-2a(P0.01)和COX-17(P0.05)基因的表达,同时0.036 mg·L-1银离子增加了斑马鱼体内p53基因的表达(P0.05)。以上研究结果说明,纳米银可能通过caspase通路诱导细胞凋亡进而影响斑马鱼胚胎的生长发育;而银离子不但影响氧化系统基因通路,还能通过p53诱导凋亡进而阻滞斑马鱼胚胎的生长发育。     

Control of larval release in the Caribbean spiny lobster, <Emphasis Type="Italic">Panulirus argus</Emphasis>: role of chemical cues

The current model for larval release in subtidal crustaceans suggests that hatching time is controlled by the embryos, which release a pheromone that stimulates the parent female to undergo behaviors that synchronize larval release. Alternatively, hatching could be controlled by the females. Ovigerous spiny lobsters Panulirus argus (Latreille) exhibit stereotypic behaviors during larval release, including rapid abdominal extensions and pleopod-pumping activity. Ovigerous P. argus were collected from coral reefs in the Florida Keys, USA during the summers of 2005 and 2006. Pleopod-pumping activity was quantified to determine if a female’s pumping activity correlates with the developmental state of the embryos. The role of pheromones released by developing and hatching embryos in controlling pumping behaviors was tested by measuring the pumping response of ovigerous lobsters to (1) hatch water, (2) homogenized embryo water, (3) embryo-conditioned water (unhatched late-stage embryos soaked for 20 h), and (4) water containing homogenized post-hatch embryo cases. Bioassays were conducted under constant conditions (dim-red light) in the laboratory at random times during the day to control for any possible rhythm in pumping activity. Spontaneous pleopod-pumping activity increased significantly with increasing embryo development. Upon exposure to hatch water, ovigerous lobsters with late-stage embryos displayed increased pleopod pumping with increased treatment concentration. Water individually conditioned with homogenized late-stage embryos, intact late-stage embryos, and homogenized post-hatch embryo cases all induced larval release behaviors in females with late-stage embryos. Ovigerous females with early-stage embryos did not respond to water conditioned with homogenized early- or late-stage embryos. These results suggest that active substances are released by embryos at the time of hatching and induce the stereotypical pumping behaviors of the female that synchronizes larval release. The results support the model that larval release in subtidal crustaceans is controlled by pheromones released from hatching embryos.     

Automated interpulse duration assessment (AIDA) in the shore crab Carcinus maenas in response to copper exposure

The effects of acute (24 h) and chronic (4␣weeks) copper exposure and subsequent recovery in “clean” conditions on cardiac activity were examined in shore crabs ( Carcinus maenas). Cardiac activity was monitored by automated interpulse duration assessment (AIDA) which permits detection of irregularities in cardiac rhythms in selected invertebrates. Mean interpulse durations, maximum interpulse durations and coefficients of variation were significantly affected by exposure to copper for 24 h and 4 weeks. Mean interpulse durations decreased reflecting an increase in heart rate, whereas maximum interpulse durations and coefficients of variation increased with increasing exposure concentration. Recovery of normal cardiac activity was evident following a 6-week recovery period in “clean” conditions. However, high levels of mortality were seen among crabs exposed to the two highest copper concentrations (0.6 and 0.8 mg l⁻¹ Cu). The extent of recovery was assessed by subjecting the surviving crabs to a physiological challenge. There were no apparent differences in circulatory response to physiological challenge between crabs previously exposed to copper and control crabs. Gill copper concentrations were significantly elevated in crabs that had previously been exposed to copper, despite their return to “clean” sea water for 6 weeks. Midgut gland wet weight/dry weight ratios, haemolymph total protein and midgut gland metallothionein concentrations did not reflect the previous copper exposure. The findings of the present study indicate that AIDA may be a valuable tool for use in toxicity testing. Received: 24 March 1997 / Accepted: 21 October 1997     

Variable within-brood maternal provisioning in newly extruded embryos of Homarus gammarus

Offspring quality of decapod crustaceans has been widely studied, with special emphasis on the sources of variability determining embryonic and larval quality. Nevertheless, maternal provisioning has commonly been overlooked as a potential source of offspring within-brood variability. In the present study, the existence of variable maternal provisioning was assessed through the analysis of the fatty acid (FA) profile of newly extruded embryos from different regions of the brooding chamber of Homarus gammarus. Significant differences in the FA profile of embryos sampled from different pleopods and sides of the brooding chamber were recorded. Significant deviations of the overall mean of each surveyed female were also observed for essential FA, particularly 20:4n-6, 20:5n-3 and 22:6n-3. Lipid energy available to fuel embryogenesis also varied among embryos sampled from different regions of the brooding chamber. Results suggest variable female investment at oocyte production, which may be amplified during the incubation period of developing embryos by differential lipid catabolism. For the first time, maternal provisioning is evidenced as an additional source for within-brood variability in the FA profile of embryos.