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.

     

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.

     

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.

     

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.

     

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.

     

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.

     

Urine composition and kidney function in the pulmonate freshwater snail Lymnaea stagnalis

• 1.1. Final urine is intermittently released from the pneumostome of the pulmonate freshwater snail Lymnaea slagnalis. A technique is described to sample this fluid.
• 2.2. The ionic composition of final urine greatly differs from that of haemolymph; Na⁺ and Cl⁻ are reabsorbed to a considerable degree. In lettuce fed snails K⁺ is excreted.
• 3.3. The urine Na⁺ and Cl⁻ concentrations are about 38 and 31 mM lower, respectively, than the haemolymph concentrations, also when the latter concentrations vary.

     

Patterns of fuel utilization by the thoracic muscles of adult worker ants. The use of lipid by a hymenopteran

• 1.1. Patterns of fuel utilization in the thoracic muscles of three species of ants have been established.
• 2.2. The thoracic muscles of Formica ulkei exhibit a typical Hymenopteran metabolic organization, relying exclusively upon carbohydrate oxidation for the provision of metabolic energy. This species feeds upon honeydew.
• 3.3. Pogonomyrmex californicus, a granivorous ant, exhibits a metabolic organization unprecedented for a Hymenopteran species. Its thoracic energy metabolism is based upon lipid oxidation.
• 4.4. Atta colombica, a fungus feeder, can metabolize both carbohydrate and fat, a versatility which is not typical of Hymenoptera.
• 5.5. It is concluded that patterns of fuel utilization in insects are not determined by phylogenetic inertia, but are selected to accommodate the activity patterns, feeding ecology and dietary regime of the species.

     

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.

     

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.

     

Dose-dependent incorporation of orally infused [1-14C]oleic acid into the lipid classes of midgut,haemolymph and fat body of dragonfly larvae (Aeshna cyanea)

• 1.1. Five different doses of radioactive oleic acid (ranging from 1.87 nmoles to 5.61 μmoles) were administered to Aeshna cyanea larvae.
• 2.2. Its incorporation into the midgut epithelium, haemolymph and fat body increased with the dose and time.
• 3.3. Low doses caused up to 95% phospholipid labelling in the midgut wall, while labelled triacylglycerol was less than 1%, but increased with the doses to a maximum of 68%. The data favour the glycerophosphate pathway of oleic acid esterification.
• 4.4. At low doses oleic acid was mainly released into the haemolymph from the midgut phospholipid pool, and at high doses from the triacylglycerol pool.
• 5.5. Diacylglycerol was the most heavily labelled lipid class of the haemolymph, amounting up to 98% and slightly decreasing with time.
• 6.6. The fat body showed a dose- and time-dependent increase in labelled phospholipid and triacyl-glycerol, maximally amounting to 14 and 90%, respectively.

     

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.

     

Osmoregulation and temperature effects on water loss and oxygen consumption in two species of african scorpion

• 1.1. Aspects of the physiology of two southern African scorpions have been examined. The scorpions are the large desert species Parabuthus villosus (Peters) (Buthidae) and the more mesic, burrowing species Opisthophthalmus capensis (Herbst) (Scorpionidae).
• 2.2. Evaporative water losses were higher in Opisthophthalmus at all temperatures.
• 3.3. Analysis of haemolymph during prolonged desiccation showed good osmotic and ionic regulation in Parabuthus but no regulation in Opisthophthalmus.
• 4.4. Oxygen consumption of Parabuthus was measured after acclimation to 10 and 30°C. Metabolic rates were extremely low but there was no metabolic compensation to increased temperatures.

     

The association of changes in cerebral ganglion dopamine metabolism with ootheca development in Periplaneta americana

• 1.1. Dopamine levels and DOPA-decarboxylase activity were measured in cerebral ganglia and haemolymph of female Periplaneta americana.
• 2.2. Measurements were made at four points in the oothecal cycle of cockroaches known to drop oothecae at regular three day intervals.
• 3.3. Dopamine levels and DOPA-decarboxylase activity in haemocytes and plasma cycle in phase with ootheca formation; their levels in haemolymph are maximal when a half visible, untanned ootheca is present.
• 4.4. In the cerebral ganglia dopamine levels and DOPA-decarboxylase also cycle in phase with ootheca formation suggesting that cerebral ganglion dopamine metabolism is under the same controls as dopamine metabolism associated with oothecal tanning.

     

Evidence for hyporegulation in the calanoid copepod,Acartia tonsa

• 1.1. The euryaline calanoid copepod, Acartia tonsa, maintains haemolymph Na below that of the external medium in salinities above 34^o_oo (475 mM Na).
• 2.2. The measured transepithelial electrical potential. −9.97 ± 1.0 mV, indicates that Na is regulated out of electrochemical equilibrium.
• 3.3. Water osmotically lost in hyporegulation is replaced by Na-dependent absorption by the gut.
• 4.4. High osmotic water permeability is evidenced by the fact that with an increase in external salinity from 475 mM Na to 580 mM Na the copepod's drinking rate nearly doubles.
• 5.5. Sodium efflux measurements indicate that ionic permeability is much lower than other hyporegulating crustaceans.
• 6.6. The energetic advantage of hyporegulation in this species is considered.

     

Ionic regulation and salinity related changes in haemolymph protein in the semi-terrestrial beachflea Orchestia gammarellus (Pallas) (Crustacea: Amphipoda)

• 1.1. Orchestia gammarellus maintained in air and provided with food in the form of agar was found to be very tolerant of changes in the ionic content of the food and was shown to have well-developed powers of ionic regulation over the salinity range 5–40‰ at 10°C.
• 2.2. There was an inverse relationship between haemolymph protein and acclimation salinity.
• 3.3. The concentration of sodium and protein ions in the haemolymph of O. gammarellus from above high water mark (H.W.M.) was markedly different from animals collected below H.W.M. Individuals taken from above H.W.M. characteristically had low haemolymph sodium but elevated haemolymph protein concentrations.

     

Development of imaginal competence to adipokinetic hormone in Locusta: Lipid and carbohydrate levels,and glycogen phosphorylase activity in azadirachtin-induced over-aged nymphs

• 1.1. Adipokinetic responses to injection of synthetic adipokinetic hormone I (sAKH) and relevant physiological parameters were studied in azadirachtin-induced 31 to 34-day-old Vth-instar over-aged male nymphs of Locusta migratoria which did not undergo the metamorphic moult to adult.
• 2.2. The resting lipid and carbohydrate levels in the haemolymph and sAKH-induced activation of fat body glycogen phosporylase did not differ markedly between over-aged nymphs, normal nymphs and normal adults.
• 3.3. Total fat body glycogen phosphorylase activity and the effect of sAKH on haemolymph carbohydrate level in over-aged nymphs were similar to those in normal adults and differed from those in normal nymphs.
• 4.4. sAKH-induced elevation of haemolymph lipid level in the over-aged nymphs was higher than in normal nymphs but lower than in normal adults; thus, the over-agad nymphs attained only a partial adult competence in this respect.

     

Neurohypophysial hormones as molecular evolutionary tracers: investigations on the sturgeons Acipenser stellatus and Acipenser guldenstadti

• 1.1. Neurohypophysial hormones of two sturgeon species, Acipenser stellatus and Acipenser guldenstadti, have been purified through molecular sieving on Bio-Gel P4 and reverse-phase high pressure liquid chromatography on Nucleosil C18 columns.
• 2.2. Arginine vasotocin has been identified in both species by its retention time in partition chromatography, amino acid composition and, in the case of A. stellatus, by amino acid sequencing.
• 3.3. A second peptide has been purified and could be α-deamidated vasotocin.
• 4.4. Another peptide with oxytocic activity, distinct from the known oxytocin-like peptides, seems to be present in very small amounts.

     

Lipids in the storage organs of three mermithid nematodes and in the hemolymph of their hosts

• 1.1. Trophosomes of the mosquito parasitic mermithid Romanomermis culicivorax and two strains of the blackfly parasitic mermithid Neomesomermis flumenalis contained in order of prevalence: triacylglycerols, phospholipids, sterols and sterol esters.
• 2.2. Triacylglycerols in the trophosomes comprised a wide array of fatty acids, sterol esters a smaller variety.
• 3.3. The degree of unsaturation of triacylglycerols and sterol esters was greater for N. flumenalis than R. culicivorax.
• 4.4. The hemolymph of the three host species (Aedes aegypti, Prosimulium mixtum/fuscum, Simlium venustum) contained an assortment of fatty acids, with a higher degree of unsaturation in the simuliids than in the culicid.
• 5.5. The overall lipid concentration of the blood of A. aegypti was unaltered by parasitism, but the mermithid caused an increase in the myristic acid:palmitic acid ratio in the free fatty acid fraction of the host's haemolymph.