Variations in protein levels in the hemolymph, hepatopancreas and ovary of Penaeus schmitti during ovarian maturation (Crustacea, Decapoda, Peneidae)

Variations of protein concentrations in the haemolymph, hepatopancreas and ovaries were studied during the ovarian maturation of the shrimp Penaeus schmitti. The main variations observed during this physiological process are the increase of protein and the decrease of water contents in the ovary, more pronounced at the first stages of gonadosomatic index (GSI:0-4). During ovarian maturation, water content of the ovaries and hepatopancreas decrease from 10.7 and 3.9% respectively. In relative values, protein content increases from about 40.0% in the ovary, 47.5% in the haemolymph and 15.4% in the hepatopancreas. In absolute values, the protein concentrations of the hepatopancreas showed no significant variation. In the opposite, ovary protein content is 14.7 fold higher at the end than the early stages of vitellogenesis. Relations between haemolymph, hepatopancreas and ovary with eventual transfers of protein material are discussed.     

Variation in the free amino acid content of ovary, hepatopancreas and hemolymph of Penaeus schmitti during ovarian maturation (Crustacea, Decapoda, Penaeidae)

Quantitative and qualitative variations of free-amino-acids contents (FAA) of ovaries, hepatopancreas and haemolymph have been studied in the course of ovarian maturation in Penaeus schmitti. In the ovary, FAA content, expressed as microM/g ovary fresh weight, decreases from 56.2 to 23.6 microM between gonadosomatic index (GSI) 0.8 and GSI 7.1. There is an increase of hepatopancreas FAA content of 33.4% till GSI 4. Haemolymph FAA content increases from 0.32 to 1.76 microM/ml, with maximal values of 3.74 microM/ml when GSI are between 3 and 4. FAA content of ovaries, expressed as microM/g of animal, is 3.2 fold higher at the end of maturation, with a main increase in the first stages of ovogenesis. Maturation process has a quantitative effect on all FAA, but, on a qualitative point of view, it concerns mainly lysine, arginine and glycine. The main variations of AA composition (expressed as percentage of total FAA) are observed in haemolymph.     

Incorporation of palmitic acid-1-14C into lipids of pharate adult tissues of Bombyx mori

Incorporation of palmitic acid-1-¹⁴C into pharate adult tissues and their lipid components of Bombyx mori was investigated. Rapid incorporation of radioactivity took place predominantly in fat body and haemolymph lipids, and partially in ovarian lipids immediately after the injection at the middle stage of pharate adult development. The major parts of the radioactivities in fat body, haemolymph and ovary were distributed in triglycerides and phospholipids, diglycerides, and triglycerides, respectively. The patterns of time course of incorporation of radioactivity into lipid components of pharate adult tissues suggest that the major form of lipid released from fat body may be diglycerides and the diglycerides in haemolymph are probably the main source of ovarian triglycerides.     

Circadian variations in hemolymph lipids and lipoproteins and in hepatopancreatic lipids of the shrimp Penaeus japonicus

Lipids content of the haemolymph and the hepatopancreas in the decapod Crustacean P. japonicus exhibits a bicircadian rhythm characterized by one maximum in the night and another one during the day. The maximal values in the haemolymph are approximately two and a half times greater (8 mg/ml) than minimal ones (3 mg/ml). Variations are less important in the hepatopancreas. A bicircadian rhythm of lipid classes in the haemolymph is observed very significantly in concentration of polar lipids and free sterols with maximal values (6.87 mg/ml and 0.59 mg/ml) and minimal values (2.63 mg/ml and 0.23 mg/ml) respectively. Polar lipids are the major lipid fractions in the haemolymph (87%). The electrophoretic behaviour of haemolymph lipoproteins is determined.     

Variation in biochemical composition during gonad maturation of the tropical abalone Haliotis varia Linnaeus 1758 (Vetigastropoda: Haliotidae)

Biochemical changes in the body components during gonad maturation of the tropical abalone Haliotis varia were investigated using wild collected specimens from the Gulf of Mannar, on the southeast coast of India. The gonadosomatic index (GSI) and the hepatosomatic index (HSI) showed negative correlations throughout the study period as well as during the progression of gonad maturation stages. The highest GSI for both the sexes were in the ripe stages followed by late maturing stages. The HSI ranged from 2.97 to 6.71 in females, and 3.55 to 5.09 in males. Among the biochemical components analysed, lipid and carbohydrate contents showed significant variations in the different tissues of H. varia during the progress of gonad maturation. The highest protein content was in the foot muscle and the lowest was in the digestive gland. Total lipids in the ovary were always higher compared with that of the testis and the values ranged from 12.60 to 26.49%, registering the highest value in the ripe ovary. Gonad carbohydrate content was lower when the lipid content was higher, suggesting the conversion of carbohydrate to lipids. The present study demonstrates the role of nutrient translocation between body parts as an essential part of the reproductive physiology of abalone.     

Daily variations of the antioxidant defense system of the lithodid crab Lithodes santolla

Several physiological processes can induce daily variations in aerobic metabolism. Lithodes santolla is a decapod crustacean of special concern since it is a sub-Antarctic species of commercial interest. The aim of this work was to study in L. santolla the daily variations in levels of enzymatic and non-enzymatic antioxidants, lipid peroxidation and protein oxidation, and haemolymphatic pH. Males of L. santolla of commercial size were randomly dissected every 4 h during a period of 24 h. Enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase and glutathione peroxidase were determined in samples of gills, muscle, hepatopancreas and haemolymph. Ascorbic acid, total glutathione, lipid peroxidation and protein oxidation were also determined in all tissues. Gills showed the highest enzymatic activity and hepatopancreas the highest concentration of non-enzymatic antioxidants. Maximum antioxidant activity was during the dark phase in gills and during the photophase in the haemolymph. Muscle showed significant daily variations, with peaks during the photophase and scotophase. Overall, an antioxidant protective mechanism is present in all tissues, where SOD and CAT represent the first line of defense. The defense mechanism in L. santolla seems to be more active during the dark phase, with slight differences among the analyzed tissues, indicating a higher metabolic rate.     

Dynamics of vitellogenin mRNA expression and changes in hemolymph vitellogenin levels during ovarian maturation in the giant freshwater prawn Macrobrachium rosenbergii

The dynamics of vitellogenin mRNA expression during ovarian maturation in Macrobrachium rosenbergii were examined by measuring hemolymph vitellogenin (Vg) levels and Vg mRNA expression in the hepatopancreas and ovary at differing reproductive stages in both intact and eyestalk ablated animals. Vg mRNA was quantified using real-time RT-PCR and hemolymph Vg was measured by enzyme immunoassay. In intact animals, Vg mRNA levels in the hepatopancreas and hemolymph Vg levels showed a gradual increase during the molt cycle concomitant with increasing gonadosomatic index (GSI), with Vg levels decreasing prior to ecdysis although GSI continued to increase. Eyestalk ablation was seen to accelerate Vg synthesis as well as ovarian maturation, although it did not alter the overall pattern of Vg expression. Vg mRNA expression was negligible in the ovary of both intact and eyestalk ablated animals, confirming that the hepatopancreas is the principal site of Vg synthesis in M. rosenbergii with the ovary being only a minor contributor. This study has shown that Vg synthesis is correlated to ovarian maturation and the molt cycle in M. rosenbergii.     

Multiplication cellulaire et production lipidique en culture in vitro chez Taphrina insititiae et Taphrina pruni, parasites de Prunus domestica

Cellular Multiplication and Lipid Synthesis during in vitro Culture of Taphrina insititiae and Taphrina pruni, Parasites of Prunus domestica. Two Taphrina species were grown on “yeast extract/glucose” and harvested at regular intervals. Their growth was estimated by dry mycelial weight and cell nitrogen. Lipids were recovered and measured by weighing. Phospholipids, which form the major portion of polar lipids, were measured by phosphorus assay. Lipidic and nitrogenous products were studied with regard to the evolution of the cultures during the phase of active growth. The cells accumulated fats, but during the phase of autolysis, fat content decreased. Production of nitrogenous components was maximal during the autolytic phase. Phospholipids varied as nitrogenous and inversely to neutral lipids. There was apparently an antagonism between nitrogen assimilation and synthesis of neutral lipids. Different functions of neutral and polar lipids may be involved, so that the former act as a reserve material and the latter represent an essential structural cell constituent. Fatty acids were analysed by gas liquid chromatography at various culture periods. Fatty acids from polar as well as those from neutral lipids exhibited great variations with regard to culture evolution. The ratio insaturated/saturated fatty acids is greater in polar lipids than in neutral lipids. These variations are discussed with regard to their consequences for the function of cell membranes.     

Nutritional contribution to females of14C-labeled male secretions transferred during mating inMenida scotti (Heteroptera: Pentatomidae)

Menida scotti (Puton) males have been shown to transfer secretions from their bulbus ejaculatorius and reservoir of ectodermal accessory gland to females by mating during hibernation. In the present study, the major components of the secretions were found to be proteins and lipids. To specify the female organ incorporating the male secretions, a radiotracer experiment in which the male secretions were labeled by [¹⁴C]valine was conducted in nine tissues of females collected in the fall and spring of the hibernation period. Relatively high radioactivities were detected in the haemolymph and the residual carcass (head, legs, air-sacs, exoskeleton, etc.) in the fall females, and in CO₂ gas evolved and carcasses in the spring females. The radioactivities in the fat body were significantly higher in the fall mating females than in the spring mating females, and vice versa in the ovary. The radioactivities in six fractions (lipids, proteins, glycogen, sugars, free amino acids and the residues) were also assayed in the five organs of females that had a relatively high radioactivity. The highest radioactivity was detected in the protein fraction of the haemolymph in fall and spring females. There were significant differences in the radioactivities incorporated into the lipid fractions of the carcass between fall and spring females.     

Effects of Cadmium on Lipid Storage and Metabolism in the Freshwater Crab Sinopotamon henanense

Since environmental effects of molecular traits are often questioned we analyze here the molecular effects of cadmium (Cd) on lipid pathways and their effects on tissues development. Lipids are an important energy source for the developing embryo, and accumulate in the ovary and hepatopancreas of decapod crustaceans. The extend of Cd affecting lipid storage and metabolism, is studied here with the freshwater crabs Sinopotamon henanense. Crabs were exposed to water-born Cd at 1.45, 2.9, 5.8 mg/l for 10, 15, and 20 days. With significantly increased Cd accumulation in exposed crabs, lipid content in hepatopancreas and ovary showed a time-dependent and concentration-dependent reduction, being at least one of the reasons for a lower ovarian index (OI) and hepatopancreatic index (HI). After 10-day exposure increased triglyceride (TG) level in hemolymph and up-regulation of pancreatic lipase (PL) activity in the hepatopancreas suggested an increased nutritional lipid uptake. However, two processes led to lower lipid levels upon Cd exposure: an increased utilization of lipids and a down-regulated lipoprotein lipase (LPL) led to insufficient lipid transport. 10-day Cd exposure also triggered the production of β-nicotinamide adenine dinucleotide 2''-phosphate reduced tetrasodium salt hydrate (NADPH), as well as to the synthesis of adenosine triphosphate (ATP) and fatty acids. With increasing exposure time, the crabs at 15 and 20-day exposure contained less lipid and TG, suggesting that more energy was consumed during the exposure time. Meanwhile, the level of NADPH, ATP and the activity of PL, LPL, fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC) activity was down-regulated suggesting an impairment of the crab metabolism by Cd in addition to causing a lower lipid level.     

Lipid composition of a plasma membrane enriched fraction of maize roots

The lipid composition of a plasma membrane enriched fraction isolated from corn (Zea mays) roots was examined. On a wt basis, the lipid: protein ratio was 1.11. Phospholipids comprised 60% of total lipids with the major phospholipids being phosphatidylcholine (62%) and phosphatidylethanolamine (21%). Free sterol was the major neutral lipid. The sterol:phospholipid molar ratio was 0.31. The fatty acid composition of the membrane was predominantly linoleic (60%) and palmitic (30%).     

Lipid storage and changes during flight by triatomine bugs (Rhodnius prolixus and Triatoma infestans)

Exceptionally large amounts of lipid are stored in flight muscles of Rhodnius prolixus and Triatoma infestans (197 and 90 μmoles glyceride glycerol per g fresh weight respectively). The bulk of this lipid is in the form of triacylglycerol.A significant decrease in the muscle lipid occurs during the first hour of flight. Over the same period there is an increase in haemolymph lipid (particularly of diacylglycerol) which is taken to indicate the use of lipid from the fat body. The carbohydrate content of muscle and haemolymph is low, so it is likely that the supply of energy for flight is provided almost exclusively by the oxidation of fat. Oleate and palmitate are the major fatty acid components of lipid from both Triatoma and Rhodnius and are probably also the major fatty acids used for oxidation.Maturation of flight ability is temporally associated with the development of flight muscle size and increase in glyceride content.     

Vitellogenin gene expression and hemolymph vitellogenin during vitellogenesis, final maturation, and oviposition in female kuruma prawn, Marsupenaeus japonicus

In penaeid shrimps, vitellogenin (VTG), the precursor of vitellin, is synthesized in the ovary and hepatopancreas and accumulated in oocytes during ovarian development. In the present study, VTG gene expression levels and hemolymph VTG levels were determined throughout ovarian development in female kuruma prawn, Marsupenaeus japonicus. Hemolymph VTG levels and VTG mRNA levels in the ovary and hepatopancreas were high during vitellogenesis, remained high until final maturation, and then decreased after oviposition. This profile suggests that VTG synthesis activity increases during vitellogenesis and decreases after oviposition. Absence of a significant increase in ovary size in final maturation suggests cessation of yolk accumulation and low activity of VTG synthesis in spite of high VTG mRNA levels. VTG mRNA levels in ovary and hepatopancreas were both highly correlated during vitellogenesis. Thus, their contribution to yolk accumulation seems to be similar. In contrast, VTG mRNA levels in the hepatopancreas increased more slowly at the start of vitellogenesis and declined more sharply after oviposition than in the ovary. This suggests a difference in the regulation of VTG synthesis between the ovary and the hepatopancreas.     

Metal binding compounds in hepatopancreas and haemolymph of porcellio scaber (isopoda) from contaminated and reference areas

1. In order to evaluate the role of metal-binding proteins in the tolerance mechanism of Porcellio scaber to heavy metals, a comparative study was made using isopods from three locations: a zinc-lead mine (Plombiéres), a zinc smelter (Budel) and a reference wood (Spanderswoud). The Cu, Zn and Cd concentrations and the protein composition were determined in the haemolymph and hepatopancreas from the isopods.2. A constant Cu/Zn molar ratio of about 5 was found in the haemolymph of all populations and no correlation was found between hepatopancreas and haemolymph Cu and Zn content.3. Using fast-performance liquid chromatography (FPLC), most of the haemolymph Cu and Zn appeared to be associated with a single UV absorbing peak corresponding with an apparent molecular weight of ± 70 kD; this peak is probably the monomer of hemocyanin.4. The hepatopancreas Zn and Cd concentration were elevated compared to the hepatopancreas of the smelter and mine isopods; after homogenization and centrifugation 70–80% of the metals were found in the supernatant.5. In all populations the hepatopancreas Cu-, Zn- and Cd-binding compounds eluted in separate peaks of low molecular weight, suggesting the absence of an MT-like compound in Porcellio scaber.6. The similarity of the protein profiles in haemolymph, and the similar distribution of the metals over the fractions in haemolymph and hepatopancreas suggests that inducible metal binding compounds are not involved in metal tolerance differences between populations.     

Lipid metabolism of the male crab Pachygrapsus marmoratus during the moulting cycle

The moult induces important variations in the concentrations and fatty acid composition of lipid classes during the moulting cycle of the male crab Pachygrapsus marmoratus. Phospholipids are the predominant lipid class. The high levels in total saturated fatty acids but also in oleic and palmitic acids result of the semi-terrestrial ethology of this grapsidae. The hepatopancreas presents the most notable variations in the lipid concentration during the moulting cycle. Transport of hepatopancreatic lipids through the hemolymph to the periphery occurs during the end of premolt, just before the new tissues constitution.     

Influence de la température sur quelques activités enzymatiques chez Palaemon serratus

Variation in Esterase 2 C activities, involving the hydrolysis of 2-carboxylic esters, α-glucosidase acetyl-glucosaminidase and alkaline and acid phosphotases, in the hepatopancreas and the abdominal muscle of Palaemon serratus was examined by polyacrylamide gradient gel electrophoresis. Soluble proteins were measured in the hepatopancreas and the abdominal muscle, and trypsin and chymotrypsin activities in the hepatopancreas. The activities and the isoenzymatic variations in shrimps acclimated at 5 different temperatures (between 14 and 30°) were compared and the molecular weight of each isozyme evaluated. It was found that: (a) the concentrations of soluble proteins decrease in the hepatopancreas between 18 and 30°, but remain unchanged in the abdominal muscle; (b) esterase and phosphatase activities increase with temperature but in a more or less random manner, according to the isozyme under consideration; (c) glycosidase activities increase with temperature; and (d) trypsin activity varies in an inverse relation to chymotrypsin activity.     

Biochemical composition of ovary, embryo, and hepatopancreas in the grapsoid crabs Armases cinereum and Sesarma nr. reticulatum (Crustacea, Decapoda)

Biochemical composition of ovary, embryo, and hepatopancreas tissues in wild populations of Armases cinereum and Sesarma nr. reticulatum were monitored during the reproductive season. Total lipid, carbon, nitrogen, C:N ratio, and water concentration of the ovary, hepatopancreas and embryos were quantified over the course of ovarian maturation. Ovary nitrogen concentration decreased as ovaries matured. Ovary lipid and carbon concentration differed significantly over the course of ovarian maturation for both species, but there was no relationship between the concentration or total content of hepatopancreas lipid and the stage of ovarian development in females. Neither species showed a relationship between measures of hepatopancreas lipid and the gonadosomatic index. There was also no simultaneously measurable net decrease in mass of the females' hepatopancreas. Lipid demands of ovarian maturation thus appear to be met in large part by increased dietary intake rather than by substantial draw down of pre-existing lipid stores from the hepatopancreas. While these temperate grapsoid crabs live with putatively fluctuating quality and quantity of food resources, no evidence could be found to demonstrate depletion of lipid concentrations in the hepatopancreas concomitant with ovarian maturation.     

Lipid composition of the fat body and haemolymph and its relation to lipid release in Oncopeltus fasciatus

Lipid composition of the fat body and haemolymph of male milkweed bug, Oncopeltus fasciatus, was determined. Triglycerides were the predominant lipids of the fat body while diglycerides accounted for the major lipid in the haemolymph. Sterols, sterol esters, and non-esterified fatty acids were present in both fat body and haemolymph besides triglycerides and diglycerides. Only traces of monoglycerides were detected.Gas chromatographic analysis of the fatty acids revealed a difference in the fatty acid composition between fat body and haemolymph glycerides and sterol esters. Oleate and linoleate were the predominant unsaturated fatty acids in both fat body and haemolymph lipids and in the milkweed seeds as well.When fat body was labelled in vivo and in vitro with ¹⁴C-palmitate, the fatty acid was incorporated largely into the triglycerides. When the prelabelled fat body was incubated with a medium containing haemolymph the fat body released lipids mainly as diglycerides. Some radioactivity was observed in the triglycerides and non-esterified fatty acids also.Electrophoretic analysis of the incubation medium containing the haemolymph revealed that the released lipids were bound to three haemolymph lipoprotein bands. Lipid mobilization, release, and transport in Oncopeltus are discussed in relation to studies on other insects.     

Influence de la température sur la composition en acides gras du muscle abdominal de Palaemon serratus

The shrimp Palaemon serratus was acclimatized at 9°, 15°, 18° and 25°, the lipid and fatty acids composition of the abdominal muscle show important variations. In relation to wet weight, total lipid level and polyunsaturated fatty acids (18:2ω6; 20:5ω3; 20:3ω6; 22:6ω3), show an inverse relationship with temperature. On the other hand, an increase of fatty acid content in relation to total lipids is observed as temperature increases. Some mono-unsaturated fatty acids have a polyunsaturated-like behaviour, others a saturated-like behaviour.     

Seasonal variations in the biochemical composition of the crayfish Parastacus defossus (Crustacea, Decapoda) in its natural environment.

The crayfish Parastacus defossus occurs in Uruguay and the state of Rio Grande do Sul in Brazil. It lives in swamps and lakes, where it digs sloped subterranean tunnels that are used as burrows. Because there is little information about the biology, physiology and ecology of this species, the aim of this study was to identify the seasonal variations of its carbohydrate and lipid metabolism. Crayfish were collected monthly (from November 2002 to February 2004) in the Lami region, Porto Alegre municipality (30 degrees 11'41'S-51 degrees 06'00'W). Haemolymph samples, used for determination of glucose, total proteins, triglycerides, total cholesterol and total lipids, were collected in the field using potassium oxalate as an anti-clotting agent. The animals and haemolymph samples were immediately frozen in the field. In the laboratory, the hepatopancreas, gills and abdominal muscles were removed for determination of glycogen, triglycerides, total cholesterol and total lipids. The findings suggest that in P. defossus, lipids are an important reserve of energy used during reproduction in both males and females; whereas glycogen may be used during periods of intense activity or environmental stress.