Ontogenetic variation in metabolism, biochemical composition and energy content during the early life stages of Farfantepenaeus paulensis (Crustacea: Decapoda: Penaeidae)

Dry weight (DW), oxygen consumption, ammonia-N excretion, proximate biochemical composition (total protein, carbohydrate, lipid, water and ash), and energy content (estimated from biochemical composition and by wet combustion) were determined in early developmental stages of cultured Farfantepenaeus paulensis. Pooled samples from embryonic, larval and postlarval stages (at 26 ± 1 °C and 34 ± 1‰) were used for measurements. The study focused on physiological and biochemical processes during transitional periods of ontogeny, such as hatching, lecithotrophic and planktotrophic stages, metamorphosis, and the attainment of a benthic existence in postlarva. DW showed higher increment between protozoea I (PZ I) and mysis I (M I) than in the next mysid and postlarval stages. Individual rates of oxygen consumption and ammonia-N excretion increased, while weight-specific rates presented significant reduction throughout development. Higher weight-specific oxygen consumption was registered in nauplius III (N III) and PZ I, following a decrease in subsequent stages. Postlarval stages PL V–VI and PL X–XII exhibited the lowest values among the stages studied. Weight-specific excretion was high in N III and protozoeal stages, with maximum values in PZ II, while the following stages were marked by lower rates. O:N ratios indicated higher protein catabolism in the stages between egg and M I and a shift to more lipid utilization close to metamorphosis. Water content was higher in the protozoeal stages and decreased afterwards. Higher percentages of protein, lipid and carbohydrate (%DW) were observed in egg and nauplius stages. Protein and lipid decreased from the egg through the naupliar and protozoeal stages, rising again in mysis stages. Lipid content (%DW) decreased in PL V–VI and PL X–XII. Lipid:protein ratios showed an increase of the importance of lipid between PZ III and M II. Carbohydrates represented a minor fraction of body composition, and ash percentages increased from egg to a maximum in PZ II, decreasing in subsequent stages. Energy content determined by wet combustion or calculated by energy equivalents presented the same trend throughout development, varying similarly to protein. Protein was the main energy contributor to body energy in all stages, while the importance of lipid was higher in egg and early naupliar stages. Trends observed in metabolic rates and body composition may be associated to morphological and behavioral changes during the early stages of penaeid development, such as the transition from herbivory to omnivory, and the adoption of a benthic existence. Different ontogenetic energy strategies contribute to succeed through such diverse type of development. Received: 4 July 2000 / Accepted: 6 December 2000     

Seasonal differences in citrate synthase and digestive enzyme activity in larval and postlarval antarctic krill, Euphausia superba

Antarctic krill maintain large population sizes despite dramatic seasonal fluctuations in food availability, but the mechanisms for this are still debated. The aim of this study was to compare seasonal differences in enzyme activity and respiration rates of larval and postlarval krill to provide insights into their overwintering strategies. Respiration rates, activity of the metabolic enzyme citrate synthase (CS), and those of the digestive enzymes laminarinase and total proteinase were measured in austral summer west of the Antarctic Peninsula, and in autumn in the southwestern Lazarev Sea. The 100-fold difference in chlorophyll a concentrations between the two studies is representative of the classic transition from a summer bloom to sparse winter conditions. Correspondingly, adult krill showed reduced respiration rates and CS activity in autumn. However, their digestive enzyme activity was significantly higher, suggesting more efficient assimilation of food at low food levels. Similar-sized larvae showed no summer-autumn differences in respiration rates and enzyme activity, supporting suggestions that they need to feed and grow year-round. However, trends in enzymatic activity varied between the larval stages measured, implying ontogenetic changes in body structure and function. Published online: 1 August 2002     

Isolation,purification and partial characterization of four digestive proteases from the purple seastar Pisaster ochraceus

An investigation of enzymatic activity in the pyloric caeca of Pisaster ochraceus collected at Post Point, Bellingham, Washington, USA, in 1982, revealed the presence of four distinct proteases. These enzymes have been partially purified and characterized and display tryptic, chymotryptic and carboxypeptidase A-like activity. The two distinctive tryptic enzymes have molecular weights of 88 000 and 25 200 daltons, respectively, as determined by reducting SDS-PAGE electrophoresis. The chymotryptic and carboxypeptidase A proteases have molecular weights of 22 800 and 34 400 daltons, respectively. The tryptic and chymotryptic enzymes are inactivated by phenylmethylsulfonyl fluoride, indicating that all are serine proteases. Both trypsins are inhibited by soybean trypsin inhibitor; however, the high molecular weight trypsin was not inhibited by tosyl-L-lysine chloromethyl ketone. The chymotrypsin was inhibited by tosyl-L-phenylalanine chloromethyl ketone. Both reagents indicate the involvement of histidine in the active site. The low molecular weight trypsin and the chymotrypsin were able to activate bovine trypsinogen to trypsin, as determined in a modified PAGE electrophoresis with trypsinogen and casein copolymerized in the gel.     

Digestive enzymes in the ontogenetic stages of the southern king crab, Lithodes santolla

The early ontogenetic stages of the sub-Antarctic king crab Lithodes santolla were analyzed for the presence and activities of a set of important digestive enzymes. The eggs and non-feeding larvae (zoea I-III, megalopa) showed high activities of esterases, phosphatases, and exopeptidases indicating the enzymatic ability to utilize endogeneous yolk reserves. SDS-PAGE showed a continuous decrease of proteins or proteids in the range of 59–81 kDa during ontogenetic development from the eggs through the zoeal stages to the first juvenile crab stage, CI. This reduction reflects the degradation of storage compounds during lecithotrophic larval development. Activities of the endopeptidases, trypsin and chymotrypsin, were low in eggs and larvae but increased significantly in the first juvenile crab stage. These enzymes typically facilitate the first steps of proteolysis in the extra-cellular spaces of the midgut gland and in the stomach. Their scarcity indicates that the larvae of L. santolla are physiologically not prepared to digest external food. This ability seems to appear first in the CI stage. Extracts of juvenile midgut glands and the gastric fluids of adults showed high activities of a variety of digestive enzymes including phosphatases, carbohydrases, as well as endo- and exopeptidases. High activities of digestive enzymes in adults may compensate for scarce food supply and rate-limiting low temperatures in the predominantly sub-Antarctic habitats of L. santolla.     

Assessment of genetic variation in lucerne (Medicago sativa L.) using protease inhibitor activities and RAPD markers

The aim of the present study is to identify and characterize lucerne lines resistance to weevil infestation. After three years of field screening for resistance to weevil infestation, 13 lines of lucerne were selected to assess the genotypic variations for lucerne weevil (Hypera postica Gyll.) at biochemical and molecular levels. Total phenols varied from 0.15 to 0.91 mg g (DM) in these genotypes. The highest trypsin (11.11 unit mg(-1) protein) and chymotrypsin (93.0 unit mg(-1) protein) inhibitors activities were recorded in G-1-02 and B-4-03 lines respectively, whereas highest alpha-amylases inhibitor activity (14.2 unit mg(-1) protein) in C-6-01. Zymogram patterns for trypsin inhibitor activity showed quantitative variations among the lines. In total 262 DNA fragments were generated when 45 deca-mer random primers were employed. Genetic variation in terms of genetic distance ranged from 0.65 to 0.85. Sequential Agglomerative Hierarchical and Nested (SAHN) clustering using the Un-weighted Pair Group Method with Arithmetic mean (UPGMA) algorithm yielded two clusters (cluster I and II) which converged at 72% similarity level. Cluster I contained most of the lines having low level of weevil infestation. High bootstrap values (>40) indicated the significance of nodes embodied in these two clusters. However, SDS-PAGE analysis of the leaf proteins of these 13 lines showed no major variations except minor difference in the protein bands of molecular weights between 14 to 20 kD.     

Ontogenetic changes in digestive enzyme activity of the spiny lobster,<Emphasis Type="Italic"> Jasus edwardsii</Emphasis> (Decapoda; Palinuridae)

Digestive enzyme profiles of puerulus, post-puerulus, juvenile and adult stages of the spiny lobster Jasus edwardsii Hutton were determined in order to identify ontogenetic changes in digestive capabilities and assess capacity to use dietary components and to exploit diets to meet nutritional requirements. Juvenile and adult lobsters exhibited a diverse range of enzymes, suggesting they can exploit varied diets. Proteolytic activity of trypsin, chymotrypsin and carboxypeptidase A (1.86–3.70 U mg^–1 digestive gland protein) was significantly higher than carbohydrase activity of amylase, -glucosidase, cellulase and chitinase (0.0014–0.38 U mg^–1 digestive gland protein), thus showing that dietary protein was more important than carbohydrate and that the lobster is carnivorous. These conclusions are consistent with other studies that found spiny lobsters to feed predominantly on crabs, bivalves, ophiuroids and sponges. Lipase activity (0.371 U mg^–1 digestive gland protein) was also relatively high, thus showing the importance of dietary lipid. Total activities (units per digestive gland) of every enzyme assayed increased significantly with lobster carapace length. There were few significant ontogenetic trends in specific enzyme activity (U mg^–1 digestive gland protein). Amylase and laminarinase specific activity was significantly higher in small lobsters than large lobsters (regression analyses, respectively, F_(1,23)=9.84, P=0.005; F_(1,11)=19.65, P=0.001), suggesting that carbohydrates including laminarin are more important in the diet of small juveniles. Trypsin, amylase and lipase activities were detected in all non-feeding puerulus stages, suggesting that feeding is not a cue for digestive enzyme production in J. edwardsii. Significant variations in total and specific activities of amylase (F_(1,3)=14.2, P=0.00; F_(1,3)=14.2, P=0.00) and trypsin (F_(1,3)=8.8, P=0.00; F_(1,3)=21.41, P=0.00) and a declining trend in lipase total activity between non-feeding puerulus stages suggests that they may be hydrolysing endogenous energy reserves to sustain their onshore swimming activity prior to settlement. Profiles suggest carbohydrate and lipid are utilised first, followed by protein. Consistently high levels of lipase in all puerulus stages (0.24–0.7 U digestive gland^–1; P>0.05) confirm the importance of lipid as a major energy substrate.Communicated by G.F. Humphrey, Sydney     

Ontogenetic diel vertical migration of the planktonic copepodCalanus sinicus in the Inland Sea of Japan

The ontogenetic diel vertical migration of the planktonic copepodCalanus sinicus was investigated in the Inland Sea of Japan in summer 1988, when the water was thermally stratified with a thermocline of ca 5 °C between 35 and 45 m. Stage-specific differences in the diel vertical migration behavior ofC. sinicus were found. Eggs were spawned primarily within the surface-waters between midnight and dawn by ascending females, and sank gradually to deeper waters until they hatched into nauplii. Non-feeding nauplius stages (NI and II) were distributed throughout the water column, but the first feeding stage (NIII) performed an ontogenetic upward migration. NIV to VI and copepodite (C) stages I to III continuously aggregated in the phytoplankton-rich euphotic layer. However, the depth of the median CI to III populations descended as stage progressed. The onset of prominent diel vertical migration took place in CIV, and the amplitude of vertical migration increased with age, being maximal in adult females (CVI). Adult males (CVI), however, remained in the layer below 20 m, and did not migrate dielly. The ecological significance of ontogenetic diel vertical migration is discussed.     

Some properties of trypsin-like proteases extracted from the seaweedCodium fragile and their purification

Two enzymes were purified from the green algaCodium fragile, collected in 1988 at Fukuoka, Japan, by batch-wise ion-exchange extraction, affinity chromatography and ion-exchange high-performance liquid chromatography (HPLC) using Boc-Ala-Ala-Pro-Arg-pNA as a substrate. The molecular weights of the enzymes were estimated as 38 000 and 39 000, using gel filtration HPLC. The enzymes had the same optimal pH range of 7 to 9 for both activities, and an exclusively hydrolyzed peptide bond on the carboxyl-terminal side of theL-arginine of peptidep-nitroanilides. The ratios of the enzymatic activity for X-Arg-pNA to X-Lys-pNA were larger than 100. The enzymes exhibited 30 times higher activity toward Boc-Ala-Ala-Pro-Arg-pNA when compared with trypsin. The activities were strongly inhibited by diisopropyl phosphofluoridate (DFP), and partially inhibited by phenylmethylsulfonyl fluoride (PMSF), benzamidine, leupeptin and antipain. The isolated enzymes were presumed to be trypsin-like serine protease from their primary substrate specificities and inactivation behavior.     

蛋白酶抑制剂对杨扇舟蛾的中肠蛋白酶作用的研究

研究了杨扇舟蛾幼虫的中肠几种主要蛋白酶活性．以14C-酪蛋白为底物测定的总消化酶活性在碱性缓冲液中最高；特异蛋白酶抑制剂研究表明：只有丝氨酸蛋白酶抑制剂对中肠蛋白酶活性有抑制作用，其它的特异蛋白酶抑制剂如半光氨酸蛋白酶抑制剂、天门冬氨酸蛋白酶抑制剂和金属蛋白酶抑制剂对中肠的酶活均无抑制作用，可以认为杨扇舟蛾的中肠蛋白酶以丝氨酸蛋白酶为主．以特异的p－nitroanilide为底物测定胰蛋白酶、胰凝乳蛋白酶、弹性蛋白酶和亮氨酸蛋白酶活性在碱性缓冲液中最高，且以弹性蛋白酶活性最高．在所研究的23个丝氨酸蛋白酶抑制剂中，抑制作用最强的是马铃薯蛋白酶抑制剂I和南瓜韧皮胰蛋白酶抑制剂．本研究丰富了对杨扇舟蛾的中肠蛋白酶种类的了解，为杨树抗虫基因构建和转化提供了依据．     

Effect of diet and paralarval development on digestive enzyme activity in the cephalopod <Emphasis Type="Italic">Robsonella</Emphasis><Emphasis Type="Italic">fontaniana</Emphasis>

This study aims at establishing the maturation of the digestive system in Robsonella fontaniana by means of measuring the digestive enzyme activity in paralarvae after hatching. Different groups of newly hatched paralarvae were fed with Artemia sp. (FA), Lithodes santolla zoea (FL) or were starved to serve as negative control (ST), the experimental period lasted 27 days after hatching (DAH). The semi-quantitative assay api^® ZYM (bioMérieux, France) was used to identify the pool of digestive enzymes present in paralarvae prior to selecting the specific enzymes acid phosphatase, total protease, trypsin and chymotrypsin for analysis. Mortality and larval protein content were also measured during the experiment. The survival of R. fontaniana hatchlings differed depending on the quality of the diet offered (ST < FA < FL), the same trend was observed with respect to paralarval protein content. Total protease and acid phosphatase activities were not related to either time of development or diet (P < 0.05). However, the activity of trypsin and chymotrypsin after DAH 10 was dependent on diet (P < 0.05). Paralarvae fed with Lithodes zoea (FL) performed better than paralarvae in the other two groups, exhibiting lower mortality, higher protein content and the highest proteolytic activity for trypsin and chymotrypsin. The results indicate that Lithodes-larvae are a better diet for R. fontaniana paralarvae than Artemia sp.     

Interaction between schools of fish and mysids

Summary A long-term daily census of 20 small patch reefs along the backreef of Tague Bay barrier reef, U.S. Virgin Islands, established that recently recruited postlarval French grunts, Haemulon flavolineatum, form mixed schools with at least two species of mysids (genus Mysidium). The formation of these taxonomically unrelated animals into socially interacting groups probably occurs because of the striking resemblance of postlarval grunts to mysids. Over a period of 5 d and as the postlarval grunts grow, these mixed associations break down, only to reform every 14–15 d as new postlarval grunts periodically recruit from the plankton. The fleeting but persistent formation of these mixedspecies schools is believed to initially provide protection to the grunts and, as the grunts grow, limited protection to the mysids. In additions, the grunts benefit trophically from the associations because they prey on the mysids.     

Effect of dissolved oxygen level on respiratory metabolism, nutritional physiology, and immune condition of southern king crab Lithodes santolla (Molina, 1782) (Decapoda, Lithodidae)

Episodes of hypoxia are common in the marine environment, and their ecological effects depend, in part, on their severity and duration. Many species of decapod crustaceans reside in areas with fluctuating oxygen regimens. Physiological mechanisms enhance the ability of these crustaceans to cope with acute episodes of hypoxia. Southern king crab, Lithodes santolla, fishery is important in the south of South America, and some data describe fishing zones with low dissolved oxygen (DO) levels (3.5 mgO₂ l⁻¹, i.e., 8.3 kPa). Our main objective was to evaluate the effect of dissolved oxygen level on respiratory metabolism, nutritional physiology, and immunological condition of L. santolla juveniles. Individual animals were exposed for 10 days to different oxygen tensions (2.1, 4.2, 8.5, 12.7, and 21.1 kPa) to quantify the oxygen consumption rate; thereafter, blood oxyhemocyanin (Hc), protein concentration, as well as hemocytes, were sampled. Freeze-dried animals were dissected, and digestive gland metabolites (glycogen, protein, glucose, cholesterol, acylglycerol, and lactate) and digestive enzyme activity (general protease, trypsin, and chymotrypsin), as well as gill lactate dehydrogenase (LDH) activity, were quantified. In the present study, Lithodes santolla showed a critical oxygen tension between 4 and 9 kPa, indicating that this crab species is more sensitive to DO than other crustacean species. Protein and Hc concentrations followed a similar pattern to that of oxygen consumption. Digestive gland glycogen and protein concentration did not change after 10 days at different oxygen exposures, but glucose, cholesterol, and acylglycerol concentrations decreased linearly and proportionally to the available oxygen in the water. As in other decapods, chymotrypsin showed over 90% of the total quantified proteases activity. Chymotrypsin activity together with total proteases and trypsin was not affected by the environmental oxygen tension. Gill LDH and digestive gland lactate followed a similar increase at lower environmental oxygen tension but dropped sharply at the lowest tension (2.1 kPa). Dissolved oxygen affected also the immune system through reduction of hemocytes. This could provide a critical window for opportunistic pathogens to become established when crabs are exposed to hypoxic conditions. L. santolla juveniles show a moderate tolerance to low oxygen availability by modifying the concentration of hemolymph proteins, mainly OxyHc, some digestive gland metabolites, and by activating the anaerobic metabolism. This allows L. santolla juveniles to inhabit temporarily low oxygen zones in the deep ocean and suggests an advantage for culture conditions.     

A new species of Penaeus (Crustacea: Penaeidae) revealed by allozyme and cytochrome oxidase I analyses

 Penaeid shrimps comprise an important portion of the world's industrial fisheries and mariculture. In the Southwest Atlantic, Penaeus (Farfantepenaeus) subtilis, P. (F.) paulensis, P. (F.) brasiliensis and P. (Litopenaeus) schmitti are the most important commercially exploited species. Despite their high commercial value, there is little information available on the different aspects of their biology or genetics and almost no data on stock structure. Also, the taxonomic status of P. subtilis populations in relation to P. paulensis and their geographic ranges have been recently questioned. In this paper we used both nuclear (allozymes) and mitochondrial (cytochrome oxidase subunit 1) data to compare species of south-western Atlantic penaeids and to estimate their phylogenetic relationships. We could readily discriminate among the four main Brazilian penaeid species and detected a fifth, new, species of Penaeus. This new species corresponded to one of the described morphotypes of P. subtilis. Based on these analyses, the geographic ranges of P. subtilis and P. paulensis were clarified. Our data also support the conclusion of Baldwin et al. (1998) that the subgenus Farfantepenaeus is polyphyletic. Received: 6 January 2000 / Accepted: 13 June 2000     

Vertical distributions of eggs,nauplii and copepodites of Calanus helgolandicus (Copepoda: Crustacea) in the Celtic Sea

The vertical distributions of eggs, nauplii, copepodites and adults of Calanus helgolandicus (Claus) from five oblique plankton-net hauls taken in May (1980), March and September (1981) and January (1982) at a site in the shelf sea to the south-west of the United Kingdom are described. The water depth is approximately 95 m and becomes thermally stratified during the summer months when a thermocline of ∼6 C° develops. In early spring when the water column was isothermal (∼8 °C), the development of the eggs and nauplii took place below 60 m and a single ontogenetic migration was observed between Nauplius VI (NVI) and Copepodite I (CI). As the temperature of the water increased, this migration occurred in progressively earlier naupliar stages. The eggs were distributed throughout the water column in the profile taken in early May when a 1 C° thermocline occurred between 30 to 40 m. The majority of the NI to NIV stages occurred below 40 m, with the ontogenetic migration taking place in the NIV stage; the NV and NVI stages were found above the thermocline. In September, the eggs were again distributed throughout the water column (101 490 m^-2), with a maximum number of >4 500 m^-3 occurring in the surface to 5 m depth interval. Nauplius I and II were found at all depths, demonstrating that hatching occurred throughout the water column. The ontogenetic migration in these late-summer profiles took place between the NII and NIII stages, the remainder of the nauplii being found above the thermocline in the top 20 m. This is the first time that an ontogenetic migration, similar to the developmental ascent observed in the naupliar stages of the euphausiid Euphausia superba in the deep ocean, has been shown for a copepod nauplius.     

Temporal patterns and transport processes in recruitment of spiny lobster (Panulirus argus) postlarvae to south Florida

We used time-series analysis to identify weekly and annual patterns in the supply of spiny lobster, Panulirus argus (Latreille, 1804), postlarvae to the Florida Keys, USA, over an 8 yr period. We also investigated the relationship between postlarval influx and wind forcing as a transport mechanism using the complex vector-scalar correlation analysis. Postlarval supply had a lunar phase periodicity at 4.5 wk intervals, with postlarval abundance peaking between the new moon and first-quarter lunar phases. A distinct annual cycle of postlarval supply with two peak periods was also apparent. Cross-correlation analysis between relative postlarval abundance and a 12 mo cycle showed that the annual peak occurs in spring, centered around March. With the 12 mo periodicity removed, a smaller peak at 5 mo intervals was also well defined. Wind-forcing for 7 d prior to the time of postlarval collection was marginally correlated with postlarval abundance through the entire time-series; the association was strongest during the late fall to early spring months. The analysis indicated that postlarval supply was correlated with winds from the northeast (ca. 45°), which are associated with winter atmospheric fronts. In contrast to results reported for other spiny lobster populations, these patterns suggest that recruitment of lobster postlarvae to south Florida is predictable only at a gross level and is presumably affected by the temporally inconsistent structure of regional oceanic gyres and variability in the timing of lobster spawning in the Caribbean. Received: 24 January 1997 / Accepted: 4 March 1997     

A protein signal triggers sexual reproduction in Brachionus plicatilis (Rotifera)

The defining feature of the life cycle in monogonont rotifers such as Brachionus plicatilis (Muller) is alternation of asexual and sexual reproduction (mixis). Why sex is maintained in such life cycles is an important unsolved evolutionary question and one especially amenable to experimental analysis. Mixis is induced by a chemical signal produced by the rotifers which accumulates to threshold levels at high population densities. The chemical features of this signal were characterized using size exclusion, enzymatic degradation, protease protection assays, selective binding to anion ion exchange and C3 reversed phase HPLC columns, and the sequence of 17 N-terminal amino acids. These studies were carried out over two years beginning in 2003 using B. plicatilis Russian strain. When rotifer-conditioned medium was treated with proteinase K, its mixis-inducing ability was reduced by 70%. Proteinase K was added to medium auto-conditioned by 1 female ml⁻¹ where typically 17% of daughters became mictic and mixis was reduced to 1%. A cocktail of protease inhibitors added to conditioned medium significantly reduced degradation of the mixis signal by natural proteases. Conditioned medium subjected to ultrafiltration retained mixis-inducing activity in the >10 kDa fraction, but the <10 kDa fraction had no significant activity. The putative mixis signal bound to an anion exchange column, eluting off at 0.72 M NaCl. These fractions were further separated on a C3 reversed phase HPLC column and mixis-inducing activity was associated with a 39 kDa protein. Seventeen amino acids from the N-terminus have strong similarity to a steroidogenesis-inducing protein isolated from human ovarian follicular fluid. The 39 kDa protein is an excellent candidate for the rotifer mixis induction signal.     

极低频电磁场对萌发期大豆的生物学效应

观察了50Hz.0.2mT或6.0mT的电磁场暴露对大豆的萌发过程，丙二醛（MDA）含量，超氧化物歧化酶（SOD），过氧化氢酶（CAT），过氧化物酶（POD），蛋白酶活性和可溶性蛋白含量以及幼苗根茎的组织形态学影响。结果表明：电磁场暴露下，大豆的开始萌发时间由32h提前到8h，萌发率到达50％的时间由72h分别提前到48h(0.2mT组）和40h(6.0mT组）；MDA含量随磁场强度的增大而下降；SOD，POD活性随磁场强度的增大而升高，而CAT活性在0.2mT暴露组下降，在6.0mT暴露组升高；蛋白酶活性和可溶性蛋白含量分别随磁场强度的增大升高或降低。幼苗根茎细胞的大小和形态没有明显变化。结果提示，极低频电磁场可以改变抗氧化酶的活性，减缓脂质过氧化，增强蛋白酶的活性，促进大豆的萌发过程。图4参6。     

Behaviour and tolerance to hypoxia in juvenile Norway lobster (Nephrops norvegicus ) of different ages

The annual occurrence of hypoxia (<25% oxygen saturation) in the bottom waters along the Swedish west coast coincides with the postlarval settlement of Norway lobster, Nephrops norvegicus (L.). This study investigates behaviour and the experimental effects of low oxygen concentrations in juvenile N. norvegicus of different ages. All experimental individuals were reared to the juvenile (postlarval) stage in the laboratory and then given sediment as a substratum. Behavioural responses to low oxygen concentrations were tested in early and late Postlarvae 1 exposed to normoxia (>80% oxygen saturation, pO₂ > 16.7 kPa), moderate hypoxia (30% oxygen saturation, pO₂ = 6.3 kPa) and hypoxia (25% oxygen saturation, pO₂ = 5.2 kPa). The experiments were run for a maximum period of 24 h or until individuals died. Behaviour was studied using sequential video recordings of four behavioural activities: digging, walking, inactivity or flight (escape swimming up into the water column). Behaviour and mortality changed with lowered oxygen concentrations; energetically costly activities (such as walking) were reduced, and activity in general declined. In normoxia, juveniles initially walked and then burrowed, but when exposed to hypoxia they were mainly inactive with occasional outbursts of escape swimming. To increase oxygen availability the juveniles were observed to raise their bodies on stilted legs (similar to adults in hypoxic conditions), but oxygen saturations of 25% were lethal within 24 h. The results suggest that the main gas exchanges of early postlarval stages occur over the general body surface. Burrowing behaviour was tested in Postlarvae 1 and 2 of different ages held in >80% oxygen saturation for 1 wk. The difference in time taken to complete a V-shaped depression or a U-shaped burrow was measured. The results showed a strong negative relationship between postlarval age and burrowing time, but all individuals made a burrow. Juveniles were more sensitive to hypoxia than adults. Thus, the possible consequences of episodic hypoxia for the recruitment of Nephrops norvegicus and for the recolonization of severely affected areas are discussed. Received: 4 August 1996 / Accepted: 11 October 1996     

Effect of ammonia on survival and osmoregulation in different life stages of the lobsterHomarus americanus

Acute toxicity of ammonia was determined for cultured larval, postlarval, and wild adult lobsters (Homarus americanus) in 1988. Ammonia tolerance was found to increase with ontogenetic development. Based on 96-h LC₅₀ values of 58 mg l^–1 NH₄ ⁺ + NH₃ l^–1 seawater (0.72 mg NH₃ l^–1) for Stage I larvae, 87 mg NH₄ ⁺ + NH₃ l^–1 (1.7 mg NH₃ l^–1) for Stage II larvae, 125 mg NH₄ ⁺ + NH₃ l^–1 (2.13 mg NH₃) for Stage III larvae, 144 mg NH₄ ⁺ + NH₃ l^–1 (2.36 mg NH₃ l^–1) for Stage IV postlarvae, 377 mg NH₄ ⁺ + NH₃ l^–1 (5.12 mg NH₃ l^–1) for adult lobsters at 5°C and 219 mg NH₄ ⁺ + NH₃ l^–1 (3.25 mg NH₃ l^–1) for adult lobsters at 20°C, recommendations for safe levels of total ammonia and un-ionized ammonia were calculated using an application factor of 0.1. Effects of ammonia on osmoregulatory capacity were studied on postlarvae and adults. Ability of postlarvae and adults to hyper-regulate in low-salinity media decreased after exposure to ammonia. In postlarval lobsters, osmoregulatory capacity was significantly affected in ammonia concentrations exceeding 32 mg l^–1. Osmoregulatory capacity in adult lobsters (5 and 20°C) was affected at 150 mg l^–1. In postlarval lobsters, a minimum exposure time of 12 h was required to impair osmoregulatory capacity. The decrease in hemolymph osmotic pressure was caused by lower hemolymph sodium concentrations. The presence of ammonia in the external medium could markedly affect the Na⁺/NH₄ ⁺ transport mechanism by permanently, temporarily, or partially impairing the transport sites for sodium.     

Chitobiase activity in the epidermis and hepatopancreas of the fiddler crab Uca pugilator during the molting cycle

The activity of chitobiase, also known as N-acetyl-β-glucosaminidase, in the epidermis and hepatopancreas of the fiddler crab Uca pugilator (Bosc, 1802), during the molting cycle, was investigated. A pH optimum of 5 to 6 was found for the enzymatic activity in both the epidermis and hepatopancreas. The temperature optimum for epidermal and hepatopancreatic chitobiase activities was 50 to 60 °C. The K _m values for epidermal and hepatopancreatic chitobiase activities at 19 °C were 0.190 ± 0.027 and 0.203 ± 0.016 mM 4-methylumbelliferyl-N-acetyl-β-glucosaminide, respectively. Hepatopancreatic chitobiase activity was significantly higher than epidermal enzymatic activity in all the molt cycle stages tested except Postmolt Stage A-B. Chitobiase activity varied significantly during the molting cycle, with the epidermal enzymatic activity in Premolt Stage D_3–4 significantly higher than in Stage C (intermolt) and Premolt Stage D₀, whereas hepatopancreatic chitobiase activity in Premolt Stage D_3–4 was significantly higher than in all other molt stages tested. The patterns of chitobiase activity in the epidermis and hepatopancreas correlate well with the hemolymph titer of ecdysteroids in U. pugilator during the molting cycle; this suggests that chitobiase activity in both tissues is regulated at least in part by the steroid molting hormones. Received: 6 May 1998 / Accepted: 12 September 1998