The effect of salinity on the osmotic,sodium and chloride concentrations in the hemolymph of euryhaline shrimp of the genus Penaeus

• 1.1. The hemolymph is isosmotic to seawater at 745 mOs/kg in P. aztecus, 768 mOs/kg in P. duorarum, 680 mOs/kg in P. setiferus, 699 mOs/kg in P. stylirostris, and 718 mOs/kg in P. vannamei.
• 2.2. The hemolymph is hyperosmotic to seawater at salinities below the isosmotic concentrations and hypoosmotic to those above.
• 3.3. With respect to sodium and chloride, the hemolymph is hyperionic to seawater at low salinities and hypoionic to seawater at high salinities.
• 4.4. P. aztecus and P. duorarum are weaker osmotic and ionic regulators at low salinities than P. setiferus, P. stylirostris, and P. vannamei.
• 5.5. There are no significant differences in the osmotic and ionic regulatory capabilities of all five species at high salinities.

     

Sex-specific gene expression in distinct tissues of the shrimp Penaeus vannamei

• 1.1. Soluble proteins extracted from male and female Penaeus vannamei tissues such as eyes, eyestalks, brain, nerve cord, hemolymph, heart, muscle, hepatopancreas, hepatopancreas membrane and cuticular epidermis were analyzed and compared by high-resolution mini-two-dimensional polyacrylamide gel electrophoresis (mini-2D-PAGE).
• 2.2. In each shrimp tissue a large number of discrete polypeptides was observed.
• 3.3. The polypeptide patterns from the same tissue of female and male shrimp were mostly similar but both qualitative and quantitative differences were noted, suggesting the presence of sex-specific gene products in various shrimp tissues.
• 4.4. Future applications of these results are discussed.

     

Size-dependent haemagglutinating activity in the haemolymph from sub-adult blue shrimp (Penaeus stylirostris stimpson)

• 1.1. The blue shrimp (Penaeus stylirostris) haemolymph is capable of agglutinating the red blood cells of several vertebrates to different titres. However, the haemagglutinin is considered non-specific because it is incapable of differentiating erythrocytes of human blood types A, B and O.
• 2.2. Haemagglutinating activity and serum protein content were determined for male and female blue shrimp ranging in size from 8.5 to 16 cm. Haemagglutinating activity decreased significantly with animal size, while protein content was unaffected.
• 3.3. The above finding is probably related to maturation of the immune system and could explain the higher susceptibility of young shrimp to parasitic and viral diseases.

     

Salinity tolerance of adult and juvenile red king crabs Paralithodes camtschatica

• 1.1. Juvenile king crabs were more tolerant of reduced salinities than adult crab; juvenile crab were better volume regulators at reduced salinities than adult crab.
• 2.2. Adult female king crab hemolymph was hyperosmotic to full seawater (30 ppt) and isosmotic to dilute seawater. Juvenile king crab (2 years old) were hypoosmotic at the same concentrations.
• 3.3. Lower osmotic concentration of juvenile hemolymph is at least partially due to lower sodium concentration.
• 4.4. Juvenile king crab can tolerate some dilution and survive for short periods in the reduced salinity of the lower intertidal zone.

     

Salt-dependent effects of juvenile hormone and related compounds in larvae of the brine shrimp,Artemia

• 1.1. Molting in state III larvae of the brine shrimp. Anemia, is interrupted, or even accelerated, when populations are exposed to various concentrations of juvenile hormone, methyl farnesoate, or methoprene in artificial sea-water.
• 2.2. The effects are believed to be salt-dependent, because exposure to these compounds in sea-water that is isotonic to larval hemolymph had no effect. This suggests that the juvenoids may target the ion transporting epithelia.

     

Ontogenic change of digestive enzymes in Penaeus monodon

• 1.1. Ontogenic changes of both proteases and carbohydrases of Penaeus monodon from different larva stages to adult were investigated.
• 2.2. Total protease activity was low during nauplius and zoea but peaked up in mysis. This was due to the activity increase of both trypsin and chymotrypsin.
• 3.3. The change of isozyme pattern of these two enzymes from different life stages of the shrimp was further determined by functional staining on an electrophoregram.
• 4.4. Activity of α-amylase increased after the post-larva stage, while that of chitinase and maltase showed a peak in zoea then gradually decreased to adult.
• 5.5. The ratio of α-amylase activity to protease coincided with the dietary change of the shrimp in different life stage.

     

Yolk synthesis in the marine shrimp,Penaeus vannamei

• 1.1. In vitro yolk synthesis was measured in fragments of the ovary of developing shrimp, Penaeus vannamei.
• 2.2. Progesterone and estradiol stimulated yolk synthesis in vitro, while ecdysterone, testosterone and estrogen had no effect.
• 3.3. A peptide factor from the eyestalks of crayfish stimulated yolk synthesis in vitro. A peptide factor from shrimp eyestalks inhibited yolk synthesis in vitro.

     

Vitellogenesis in the shrimp,Penaeus vannamei: in vitro studies of the isolated hepatopancreas and ovary

• 1.1. Yolk proteins were isolated from ovaries of the shrimp Penaeus vannamei and used as an antigen for antibody production in rabbits.
• 2.2. Protein synthesis was measured for both the hepatopancreas and the ovary in vitro, and proteins present in both tissues were immunoreactive with the antibodies.
• 3.3. Extracts of shrimp eyestalks inhibited in vitro protein synthesis by both tissues. The inhibitory factor from the eyestalks was heat stable and had a molecular weight of 3300 daltons.

     

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.

     

Haemolytic activity in the brown shrimp (Penaeus californiensis holmes) haemolymph

• 1.1. Natural haemolytic activity in brown shrimp (Penaeus californiensis) haemolymph was detected using mouse erythrocytes as target cells. This activity is unrelated to agglutinating and phenoloxidase activity, but it is another probable component of the shrimp defence system.
• 2.2. The haemolytic reaction is time and dose dependent, and a serine-protease is involved.
• 3.3. The haemolytic factor is thermolabile and has an apparent molecular weight of 23.5 kDa.

     

A comparative study on the biological properties of venoms from juvenile and adult common tiger snake (Notechis scutatus) venoms

• 1.1. The biological properties of venoms from juvenile and adult common tiger snakes (Notechis scutatus) were compared.
• 2.2. The lethality, procoagulant activity and enzymatic activities of the juvenile venom were not substantially different from those of the adult venom.
• 3.3. Electrophoretic studies, however, indicated some minor differences in the protein composition of the juvenile and adult venoms.

     

Vitellogenin synthesis in female larvae of the gypsy moth,Lymantria dispar (L.): suppression by juvenile hormone

• 1.1. Fat body from feeding-phase, last instar gypsy moth females incorporates l-[³⁵S]methionine in vitro into two vitellogenins with the same molecular masses (165 and 180 kDa) as the apo-vitellogenins found in teh hemolymph and the apo-vitellins in teh eggs.
• 2.2. Both apo-vitellogenins are observed in the medium of fat body cultures, but only the 180 kDa apo-vitellogenin is observed in extracts of cultured tissue.
• 3.3. Synthesis and accumulation of the apo-vitellogenins are suppressed in a dose-dependent manner by topical treatment with the juvenile hormone analog, methoprene, prior to day 4.
• 4.4. This suppression suggests that a declining juvenile hormone titre is involved in the initiation of vitellogenin synthesis.

     

Hemolymph ferritin and radula structure in the limpets Patelloida alticostata and Patella peronii (Mollusca: Gastropoda)

• 1.1. The mean concentration of total hemolymph iron was 3060μg/100 ml in Patella peronii and 2950μg/100 ml in Patelloida alticostata.
• 2.2. Ferritin was found to act as a major iron-binding protein in the hemolymph of both P. peronii and P. alticostata.
• 3.3. P. alticostata ferritin has a molecular weight of approximately 505,000, while that of P. peronii has a mol wt. of approximately 520,000.
• 4.4. The lateral radula teeth of both species are mineralized by deposits of silica (SiO₂) and iron in the form of goethite (α-FeOOH).
• 5.5. Hemolymph ferritin is suggested to act as a high capacity transport system to supply iron to the mineralizing front of the radula.

     

Effect of salinity on the osmotic,chloride, total protein and calcium concentrations in the hemolymph of the prawn Peneaus monodon (fabricius)

• 1.1. Osmolality and chloride concentrations in the hemolymph of Penaeus monodon became stable 1 day after molting in 32 ppt, while total protein and calcium concentrations remained stable throughout the molting cycle. When intermolt (≥ 36 hr postmolt) animals were transferred from control (32 ppt) to experimental (8–40 ppt) salinities, osmolality, chloride and total protein, but not calcium, concentrations in the hemolymph achieved steady state values 24–48 hr after transfer.
• 2.2. The hemolymph osmolality was a linear function (slope = 0.28) of medium osmolality at salinities between 8 and 40 ppt. It was isosmotic to seawater at 698 mOsm (10 g prawns) and 752 mOsm (30 g), and was hyperosmotic to the medium below isosmotic concentrations, and hypoosmotic to those above.
• 3.3. Hemolymph chloride concentration was isoionic to seawater at 334 mM, and was hyperregulated below isoionic concentrations, and hyporegulated to those above.
• 4.4. P. monodon maintained its hemolymph calcium concentration between 6.4 and 10 mM when medium salinities increased from 8 to 40 ppt.
• 5.5. Total protein concentration in the hemolymph was independent of medium salinity (8–40 ppt) and hemolymph osmolality (540–850 mOsm).

     

Ontogenetic change in thermal tolerance of the toad Bufo woodhousii fowleri

• 1.1. Premetamorphic Bufo woodhousii fowleri tadpoles exhibit a high critical thermal maximum (CTM) of 42.5°C which decreases during metamorphosis to 37°C.
• 2.2. The CTM of progressively larger (older) postmetamorphic or juvenile toads gradually increases until the adult CTM of 41.1°C is attained.
• 3.3. This gradual increase in CTM from juvenile to adult toad suggests that the physiological systems underlying thermal tolerance change or mature following metamorphosis until the adult condition is reached.
• 4.4. Basking in juvenile toads elevates their metabolic rate and thus may facilitate the acquisition of the adult physiological state.

     

The effect of salinity on respiratory metabolism in selected ontogenetic stages of the freshwater shrimp Macrobrachium olfersii (Decapoda,palaemonidae)

• 1.1. The effect of acute salinity exposure (0, 7, 14, 21, 28 and 35%.S) on the respiratory metabolism of selected ontogenetic stages (zoeae, postlarvae and adults) of the freshwater shrimp Macrobrachium olfersiiwas examined.
• 2.2. Metabolic rates are salinity independent from 14 to 28%. S in zoeae 1–4, but tend to increase with increasing salinity in zoeae 5 and 8. Postlarvae exhibit maximal rates in midrange salinities while in adult shrimps, oxygen consumption rates decrease with salinity increase.
• 3.3. Salinity has little effect on the metabolism-weight relationship, regression analysis indicating that b varies from 0.69 in 0%. S to 0.62 in 35%. S.
• 4.4. Data are discussed as to whether larval responses reflect adaptation to the adult biotope and whether development of the larval neurosecretory system might affect metabolic response to salinity exposure.

     

Purification and characterization of vitellogenin from the American cockroach,Periplaneta americana

• 1.1. Vitellogenin (VG) was isolated and purified from the hemolymph of female American cockroaches.
• 2.2. The purification method used in this study comprises two steps: the first step is based on the method originally developed for purifying lipophorin from hemolymph, and the second step is the separation of VG from lipophorin by a KBr density gradient ultracentrifugation.
• 3.3. The purified VG was characterized according to molecular weight, substructure, shape and size, and lipid composition.
• 4.4. The VG molecule is almost globular in shape with the diameter of about 15.5 nm and is indistinguishable from lipophorin in shape and size.
• 5.5. The native molecular weight determined by light scattering method was 560 kDa.
• 6.6. The VG consists of four subunits with molecular weights of approximately 102, 81, 49 and 40 kDa, respectively.
• 7.7. VG is a lipoprotein and comprises 92% protein and 8% lipid.
• 8.8. Major lipid components were found to be diacylglycerol (25%) and phospholipids (71%).

     

Induction of locomotor behavior in the giant African snail,Achatina fulica

• 1.1. The locomotor-inducting factor of the giant African snail, Achatina fulica, was examined.
• 2.2. Snails showed nocturnal circadian behavior in relative humidity at least over 50%. Although the rhythmicity was independent of light and darkness, it was disturbed easily by hydration, and hydrated snails continued to locomote throughout the day. For induction of locomotor behavior, relative humidity over 50% was the fundamental factor and water is shown to be the limiting factor for the endogeneous circadian oscillator.
• 3.3. The integument of snails showed a higher water permeability. Through the integument, hemolymph osmolality changed easily according to hydration and dehydration from about 120 to 400 mOsm/kg H₂O. Circadian behavior was induced in snails in which hemolymph osmolality ranged from about 130 to 230 mOsm/kg H₂O.
• 4.4. By hydration, hemolymph osmolality in quiescent and estivated snails which have higher osmolality decreased gradually and then they began to locomote according to the degree of dilution, and vice versa. The induction of behavior in these snails was controlled by low hemolymph osmolality.
• 5.5. Together with the endogeneous rhythmicity, water environment was shown to be the key factor for the induction of locomotor behavior.
• 6.6. Based on these results, the mechanisms of the induction of locomotor behavior in terrestrial pulmonates are proposed.

     

Organ specific changes in energy metabolism due to anaerobiosis in the sea mussel Mytilus edulis (L.)

• 1.1. Anaerobic energy metabolism was investigated in different organs of Mytilus edulis and the whole animal.
• 2.2. Succinate accumulates to high levels in most organs but remains low in the hemolymph.
• 3.3. After 16 hours propionate accumulation is observed in all organs. Experimental evidence is not sufficient yet to point out organs that produce more propionate than others.
• 4.4. Acetate is a minor end product.
• 5.5. Acetate and propionate are found in the hemolymph in amounts equal to those in the organs.
• 6.6. Animals incubated in oxygen-free seawater accumulate more end products than animals exposed to air, in the form of volatile fatty acids that are excreted into the incubation water.
• 7.7. Alanine and glutamine increase in the posterior adductor muscle. Aspartate decreases in the total animal, posterior adductor muscle and gills, while in the hemolymph decrease in alanine, asparagine, serine, threonine and proline are observed.

     

Na+-K+ ATPase and carbonic anhydrase activities in larvae,postlarvae and adults of the shrimp Penaeus japonicus (Decapoda,Penaeidea)

• 1.1. Activities of Na⁺-K⁺ ATPase and carbonic anhydrase were measured through the early post-embryonic development of Penaeusjaponicus. In adults, only the Na⁺-K⁺ ATPase activity was measured.
• 2.2. ATPase activity was variable in the successive development stages. From zero in nauplii, the activity slightly increased in zoeae, and rose sharply in mysis stages 2 and 3.
• 3.3. A further significant increase in activity was noted at the transition from late mysis to early postlarvae, concomitant with a change from the larval osmoconforming pattern of osmoregulation to the postlarval and adult hyper-hyporegulating pattern.
• 4.4. The activity of Na⁺-K⁺ ATPase, measured in isolated cephalothorax, increased from PL3 to PL4 to its maximum value in PL5; at this stage, osmoregulatory capacity was fully efficient.
• 5.5. In young stages of P. japonicus, the variations in Na⁺-K⁺ ATPase activity appear correlated with the development of osmoregulatory ultrastructures, and with osmoregulation and salinity tolerance.
• 6.6. These results are discussed with regard to their ecological and physiological implications.
• 7.7. In adults, the activity of Na⁺-K⁺ ATPase was high in gills and epipodites and no activity was detected in branchiostegites. These results are related to the ultrastructure of these organs.
• 8.8. The activity of carbonic anhydrase did not change significantly in larval and postlarval stages.
• 9.9. From these results, it is proposed that the effector sites of osmoregulation are located in branchiostegites, pleurae and epipodites in postlarvae, and in epipodites and mainly in gills in adults.