Biochemical aspects of Balanus hameri haemolymph,together with some comparative haemolymph data for Balanus balanus and Lepas anatifera

• 1.1. The concentration of protein in the haemolymph of Balanus hameri ranged from 2.0 to 17.3 mg/ml, and the lipid from 1.4 to 7.7 mg/ml; the haemolymph protein and lipid levels increased significantly prior to cross-fertilization.
• 2.2. The protein and lipid concentrations in Balanus balanus haemolymph were 8.1 and 1.7 mg/ml respectively.
• 3.3. The lipid concentration of Lepas anatifera haemolymph was 1.2 mg/ml.
• 4.4. The neutral lipid and phospholipid components of B. hameri and L. anatifera haemolymph were the same, with the major components of the phospholipid fraction being phosphatidyl ethanolamine and phosphatidyl choline.
• 5.5. The osmolarity (970.4 mOsm), chloride ion concentration (501.3 m-eq/l) and pH (7.29) of B. hameri haemolymph were also determined.

     

Effect of developmental derangements on the proteolytic and protease-inhibitory activities in Galleria mellonella (Insecta)

• 1.1. Protease inhibitory activity in the whole body homogenates of Galleria mellonella larvae exhibits maxima at the beginning of the last larval and pupal instars. Injury, chilling, immobilization, and ligations of larvae cause an increase of inhibition.
• 2.2. The inhibitory activity is high in the haemolymph but low in midgut and faty body. By contrast, the proteolytic activity is low in haemolymph and high in both midgut and fat body.
• 3.3. Starvation and ligations cause a dramatic fall of the proteolytic activity and increase of the inhibitory activity in examined organs.

     

Palmitic acid metabolism in hepatopancreas of the freshwater shrimp Macrobrachium borellii

• 1.1. The palmitic acid fate as substrate for the synthesis of either glycerides or other fatty acids was studied in vivo and in the microsomal fraction from hepatopancreas of Macrobrachium borellii.
• 2.2. Most of the palmitic acid administered in vivo circulated to the hepatopancreas, being incorporated mainly in the triacylglycerol (TG) fraction.
• 3.3. Palmitic acid transformations into palmitoleic, stearic and oleic acids were observed in the hepatopancreas.
• 4.4. The in vitro biosynthesis of TG in hepatopancreas was more active than in other tissues. In the microsomal fraction, palmitic acid was also incorporated mainly in TG, and followed the α-glycerophosphate pathway.

     

Anaerobic metabolism in sublittoral living Mytilus galloprovincialis in the mediterranean—IV. Role of amino acids in adaptation to low salinities during anaerobiosis and aerobiosis

• 1.1. When Mytilus galloprovincialis were transferred from 38 to 19%. sea water (S), the metabolism became anaerobic for at least 8 hr. After 24 hr the animals were entirely aerobic again.
• 2.2. Upon transfer to 19%. S, the total free amino acid concentration in haemolymph doubled within 4 hr, remaining nearly constant thereafter, up to 48 hr.
• 3.3. In the posterior adductor muscle a strong decrease of alanine and glycine occurred at 48 hr exposure to 19%. S, and a smaller decrease of glutamate; taurine remained relatively constant. When transferred again to 38%. S after 14 days, a strong overcompensation occurred in the concentrations of alanine and proline, and a smaller overcompensation in those of threonine and serine.
• 4.4. In the gill no distinct change in the amino acid pool occurred during 14 days of exposure, with the exception of a decrease in serine. When transferred again to 38%. S, a strong overcompensation occurred in alanine, proline, glycine and serine, and a smaller in glutamate and threonine.
• 5.5. No evidence for anaerobic metabolism in the decrease of the amino acid pool was found.
• 6.6. M. galloprovincialis is less able to adapt to low salinities than the more euryhaline M. edulis.

     

Metabolic utilization of leucine in a fat and a lean line of chicken

• 1.1. In vivo incorporation into body lipids and breast muscle proteins from l-[U-¹⁴C]leucine was studied in genetically lean or fat male chickens, fed or starved, 1 or 24 hr after intraperitoneal injection.
• 2.2. Lipogensis and portein synthesis from labelled leucine were significantly higher in fat chickens than in lean birds, particularly in those in the fed state.
• 3.3. Radioactivity in the free amino acid pool was greater in fat birds irrespective of the nutritional state.
• 4.4. However, utilization of injected l-[U-¹⁴C]leucine for lipogenesis was no more than 2%.

     

Water loss and haemolymph osmolarity of Potamon potamios,an aquatic land crab,under stress of dehydration and salinity

• 1.1. Small crabs survived over 18% water loss and large crabs 21% when in dry air. Size, temperature and relative humidity affected this rate.
• 2.2. Haemolymph osmolarity of newly collected crabs ranged from 530 to 630 mOsm/kg, depending on their size and the season.
• 3.3. When dehydrated, haemolymph osmolarity rose to over 700 mOsm/kg, and ion concentration increased by over 10%.
• 4.4. Crabs survived in sea-water for at least two weeks. Haemolymph osmolarity rose and ion concentration increased. The acclimation pattern affected the haemolymph osmolarity.

     

Cellular and intracellular localization of catalase and acid phosphatase in the midgut of Lumbricus terrestris L.: A cell fractionation study

• 1.1. Cellular and intracellular localization of catalase and acid phosphomonoesterase in the midgut of Lumbricus terrestris was studied by use of tissue fractionation.
• 2.2. At least 60–70% of the catalase resides in the chloragocyte cytosol and the remaining 30–40% resides in gut epithelium peroxisomes.
• 3.3. One of the main functions of the chloragocyte catalase is probably scavenging for H₂O₂ arising from the interaction between blood heme-protein and oxygen.
• 4.4. A simple method for the histochemical detection of cytosol catalase is proposed.
• 5.5. About 10% of the gut acid phosphatase resides in chloragocyte lysosomes. The chloragosomes contain no acid phosphatase.

     

Accumulation,distribution and toxicity of selenium in the adult house fly,Musca domestica

• 1.1. Accumulation and distribution of dietary Se in relation to mortality was investigated in adult house flies.
• 2.2. The midgut preferentially accumulated Se and thereby limited toxicity.
• 3.3. Midgut Se concentrations were from 6- to 107-fold higher than in carcass, and from 15 to 71% of the total Se was associated with midgut.
• 4.4. When dietary levels of Se were raised the midgut saturated at 15 μg Se/g tissue, followed by a rise in carcass levels to greater than 0.5 μg Se/g tissue and increased mortality.
• 5.5. Se levels in lysosomal fractions were from 3- to 50-fold higher than in other subcellular fractions, suggesting that Se is sequestered in lysosomes.
• 6.6. Se added to drinking water was toxic at 4–8 ppm.

     

Physiological pattern and further characterization of the haemolymph violet carotenoprotein of the fly Rhynchosciara americana

• 1.1. An electrophoretic purification procedure for the haemolymph violet carotenoprotein of R. americana was described. The purified protein was used for obtaining a specific antiserum.
• 2.2. This carotenoprotein contains: (1) a high weight percentage of glutamic acid, threonine and proline and a low weight percentage of histidine; (2) mannose and/or glucose as suggested by the interaction with concanavalin A; (3) phosphoryl groups.
• 3.3. The concentration of the violet carotenoprotein in the haemolymph is approximately constant during all the life cycle of R. americana.
• 4.4. The haemolymph of four species of Rhynchosciara genus shows the presence of proteins immunologically related with the R. americana violet carotenoprotein.

     

The effect of eyestalk ablation on growth,haemolymph composition and gill Na+, K+-ATPase activity of Penaeus monodon juveniles

• 1.1. Eyestalk unablated and unilaterally ablated Penaeus monodon juveniles had survival rates after 5 months of 75–72.5 and 67.5–60%, respectively.
• 2.2. Unilaterally ablated shrimps had significantly higher (P < 0.05) growth rate than unablated shrimps.
• 3.3. Eyestalk-ablatement resulted in a decrease in the haemolymph sodium concentration and an increase in the potassium and calcium concentration of shrimps.
• 4.4. The osmolarity of haemolymph and total protein concentration of unablated shrimps were demonstrated to be higher than those of unilaterally ablated shrimps.
• 5.5. The eyestalk-ablated shrimps possess higher total ATPase and Na⁺,K⁺-ATPase activities in the gill than those of unablated shrimps.

     

Gut microflora can modify fatty acid composition in liver and egg yolk lipids of laying japanese quail (Coturnix coturnix japonica)

• 1.1. The influence of the gut microflora on lipid metabolism was investigated in germ-free (GF) and conventional (CV) laying Japanese quail.
• 2.2. Serum and egg yolk cholesterol concentrations showed comparable values in both GF and CV environments.
• 3.3. The fatty acid compostion of liver lipids was modified by the presence of gut microflora. Notably, in the presence of the gut microflora, proportion of oleic acid was reduced and conversely, stearic and linoleic acids were enhanced.
• 4.4. In egg yolk lipids, the proportion of myristoleic and palmitoleic acids was significantly lowered and that of stearic acid was significantly enhanced by the presence of the gut microflora, though the difference was very small.
• 5.5. It was suggested that oleic acid could be easily either hydrogenated to stearic acid or desaturated to linoleic acid by the action of the gut microflora in Japanese quail.

     

Effects of starvation and a carbohydrate-rich diet on glycogen metabolism in a gastropod mollusc,Megalobulimus oblongus

• 1.1. Administration of a carbohydrate-rich diet increased haemolymph glucose levels and glycogen concentration in hepatopancreas, mantle and muscle.
• 2.2. Glycogen concentration in tissues decreases after 2 weeks of starvation and haemolymph glucose levels did not change significantly.
• 3.3. However, starvation did not induce a decrease in the intrinsic synthetic capacity in tissues.
• 4.4. Glycogen synthesis in tissues from animals fed with lettuce or a carbohydrate-rich diet, increases with increasing glucose concentration in the media.
• 5.5. However, in mantle slices from snails adapted on a carbohydrate-rich diet, the glycogen synthetic capacity was lower than in slices from snails fed with lettuce.

     

Effect of JH and PII on the protein content of haemolymph and ovaries of Spilostethus pandurus

• 1.1. An SDS-PAGE study of the qualitative and quantitative differences in protein bands from haemolymph and ovaries of Spilostethus pandurus females treated with JH or chemically allatectomized with precocene II, has been done.
• 2.2. The SDS-PAGE study of haemolymph revealed the occurrence of three female-specific proteins. In the ovary appeared three protein fractions (A, Band C) with mol. wts similar to those from haemolymph.
• 3.3. The three female-specific proteins from the haemolymph, and the ovary bands B,C and D were absent in the samples from PII-treated females.
• 4.4. JH accelerates ovary growth and the relative amounts of bands B, C and D were in relation to the physiological stage of the considered ovaries.

     

Characterization of an acid phosphatase—Glycoprotein from barley seeds

• 1.1. An acid phosphatase from barley seeds germinated for 120 hr, that is not affected by the presence of cycloheximide, is a glycoprotein.
• 2.2. The enzyme has an optimum pH 4.9 and a mol. wt of 57,000 as determined by SDS-gel electrophesis.
• 3.3. The main sugar is galactose amounting to one half of the protein and it is as either mono or disaccharide.
• 4.4. Serine and glycine predominate and hydroxyproline is absent.

     

Haemolymph magnesium levels in some oceanic crustacea

• 1.1. Data are presented for the haemolymph magnesium levels of a number of amphipod and decapod species from oceanic mid-waters of the northern South Atlantic.
• 2.2. The values are compared with neritic brachyuran haemolymph magnesium levels.
• 3.3. No obvious correlation was apparent between the presumed general activity levels of the oceanic Crustacea and their haemolymph magnesium concentrations; non migrating forms had not dissimilar concentrations to those species known to undertake substantial daily vertical migration.

     

An anticoagulant solution for haemolymph collection and prophenoloxidase studies of penaeid shrimp (Penaeus californiensis)

• 1.1. An anticoagulant solution was designed from data on osmolality, ionic concentration and pH to resemble shrimp haemolymph.
• 2.2. This Shrimp Salt Solution (SSS) prevents coagulation and prophenoloxidase system activation during the extraction of shrimp haemolymph.
• 3.3. The location of the the proPO system in the brown shrimp (Penaeus californiensis) was determined using this anticoagulant solution.

     

The haemocytes and the activities of leucine aminopeptidase and alkaline phosphatase during development of Ceratitis capitata: Specific secretion of a leucine aminopeptidase isozyme

• 1.1. The types of haemocytes during larval development were studied.
• 2.2. The developmental profile of leucine aminopeptidase and alkaline phosphatase was studied. The maximum LAP activity was found to be in early larval development, while the maximum alkaline phosphatase during the white pupal stage.
• 3.3. These activities were compared with those determined in cell-free haemolymph.
• 4.4. Both hydrolytic enzymes have been found histochemically in the prohaemocytes and in the plasmatocytes.
• 5.5. In cultured haemocytes experiments it was found that 64% of the total LAP activity was secreted into the incubation medium, while electrophoretic analysis of released LAP activity demonstrated that only LAP A isozyme was secreted.
• 6.6. Based on the above results we suggest that both hydrolytic enzymes are functionally important throughout larval development.

     

Induction kinetics of immune antibacterial proteins in pupae of Galleria mellonella and Pieris brassicae

• 1.1. Pupae of Galleria mellonella and Pieris brassicae given an injection with live, non-pathogenic Enterobacter cloacae or abiotic foreign molecules induce an acquired immunity that corresponds with the synthesis of haemolymph proteins of antibacterial activity.
• 2.2. This humoral defensive response which persists for several days, differs quantitatively between insect species and between the inducers used, although very different foreign bodies induced the same immune proteins in both lepidopteran insects.
• 3.3. A stronger and longer lasting response was consistently noticed in pupae immunized with non-pathogenic bacterium than after sterile nutrient broth injections.
• 4.4. A demonstrably elevated activity of haemolymph lysozyme and trace activity of cecropins found in pupae of Galleria treated with saline W, a salt solution physiological to moths, disappear soon after 36 hr from injection.
• 5.5. In P. brassicae, however, sterile insect Ringer can give a varying, if present at all, immune response.
• 6.6. A mechanical injury (sterile wounding of insect body) can occasionally induce a similar but much weaker response.
• 7.7. The antibacterial activity was drastically reduced in Pieris or completely depressed in most pupae of Galleria when actinomycin D or cycloheximide was given at an early time post-immunization with E. cloacae.
• 8.8. It is concluded that the de novo synthesis of ribonucleic acid and immune proteins is required for expression of antibacterial activity in pupal haemolymphs.
• 9.9. The synthesis of an immune mRNA was completed about 7 hr after the injection of the immunizing bacteria.

     

Characterization of Colorado potato beetle lipophorin: A hydrocarbon-rich diacylgycerol-poor lipophorin

• 1.1. The lipid composition of lipophorin from the Colorado potato beetle, Leptinotarsa decemlineata Say, was analyzed.
• 2.2. This insect lipophorin contains 44% lipid and is characterized by large amounts of hydrocarbons and small amounts of diacylglycerol.
• 3.3. This is the first observation of a diacylglycerol-poor insect lipophorin in haemolymph.
• 4.4. Since the main energy source for flight in the Colorado potato beetle is proline, the low diacylglycerol content in lipophorin must be related to its peculiar flight metabolism.
• 5.5. This lipophorin, however, can still take up appreciable amounts of diacylglycerol from the locust fat body. Hydrocarbon uptake by this lipophorin was also demonstrated.
• 6.6. The main function of this lipophorin therefore seems to be transport of hydrocarbons from oenocytes to the cuticle.

     

Amino acid synthesis during hyperosmotic stress in penaeus aztecus postlarvae

• 1.1. Glycine, proline, and taurine are the quantitatively most important amino acid osmolytes in Penaeus aztecus postlarvae.
• 2.2. Taurine dominates the amino acid pool in low salinity, while proline dominates the amino acid pool at higher salinities.
• 3.3. Although not major contributors to the pool, glutamate and alanine are constitutively synthesized from [¹⁴C]glucose and [¹⁴C]glutamate under constant salinity and under hyperosmotic stress treatments.
• 4.4. Proline synthesis from [¹⁴C]-precursors is apparent under constant high (but not low) salinity and is significantly induced by hyperosmotic stress.
• 5.5. No appreciable glycine synthesis was observed from [¹⁴C]glucose or [¹⁴C]glutamate under any experimental conditions.