The scheduling of spawning with the moult cycle in Northern krill (Crustacea: Euphausiacea): a strategy for allocating lipids to reproduction

Summary

Euphausiids moult and grow throughout their life, which implies sharing of resources between growth and reproduction for adult krill. In the Northern krill, Meganyctiphanes norvegica (M. Sars), female krill produce eggs cyclically. Spawning moult cycles alternate with vitellogenic moult cycles for lipid yolk accumulation. Histology shows that lipids are associated with the R cells of the digestive gland in both sexes, with the yolk platelets of mature oocytes and with the fat body cell membranes and blood lacunae in reproducing females. Mature female krill can have a total lipid content twice as high as males, mostly due to accumulation in the ovary, the fat body and the haemolymph. In contrast, in males, as well as in non-reproducing females, the highest percentage of lipids is found in the digestive gland and the haemolymph. In Meganyctiphanes norvegica, the most abundant lipid fractions are polar lipids and triglycerides, the latter being relatively low in reproducing female gonad and fat body. Triglycerides are believed to be a pure energy source and polar lipids are essential for membrane development in embryos. The fatty acid content and composition of the triglyceride and polar lipid fractions in females are different from males, related to both reproductive and dietary processes. Higher levels of polyunsaturated fatty acids (PUFA) in the polar lipid fraction were found in reproductive females. During the non-reproductive season, the converse was found, indicating the specific role PUFA and other fatty acids play in growth and egg production. Adaptive processes linked to reproduction were studied comparatively in three populations of the Northern krill—Clyde Sea (W, Scotland), Kattegat (E, Denmark), Ligurian Sea (Mediterranean)—all differing considerably in climatic and trophic conditions. Such adjustments in lipid synthesis and storage are viewed as reproductive strategies developed by the Northern krill in response to different environmental conditions.     

Nitric oxide signalling: insect brains and photocytes

The success of insects arises partly from extraordinary biochemical and physiological specializations. For example, most species lack glutathione peroxidase, glutathione reductase and respiratory-gas transport proteins and thus allow oxygen to diffuse directly into cells. To counter the increased potential for oxidative damage, insect tissues rely on the indirect protection of the thioredoxin reductase pathway to maintain redox homoeostasis. Such specializations must impact on the control of reactive oxygen species and free radicals such as the signalling molecule NO. This chapter focuses on NO signalling in the insect central nervous system and in the light-producing lantern of the firefly. It is shown that neural NO production is coupled to both muscarinic and nicotinic acetylcholine receptors. The NO-mediated increase in cGMP evokes changes in spike activity of neurons controlling the gut and body wall musculature. In addition, maps of NO-producing and -responsive neurons make insects useful models for establishing the range and specificity of NO's actions in the central nervous system. The firefly lantern also provides insight into the interplay of tissue anatomy and cellular biochemistry in NO signalling. In the lantern, nitric oxide synthase is expressed in tracheal end cells that are interposed between neuron terminals and photocytes. Exogenous NO can activate light production and NO scavengers block evoked flashes. NO inhibits respiration in isolated lantern mitochondria and this can be reversed by bright light. It is proposed that NO controls flashes by transiently inhibiting oxygen consumption and permitting direct oxidation of activated luciferin. It is possible that light production itself contributes to the restoration of mitochondrial activity and consequent cessation of the flash.     

Role of nitric oxide and mitochondria in control of firefly flash

In light-producing cells (photocytes) of the firefly light organ,mitochondria are clustered in the cell periphery, positionedbetween the tracheolar air supply and the oxygen-requiring bioluminescentreactants which are sequestered in more centrally-localizedperoxisomes. This relative positioning suggests that mitochondriacould control oxygen availability for the light reaction. Wehypothesized that active cellular respiration would make theinterior regions of the photocytes relatively hypoxic, and thatthe "on" signal for production of bioluminescence might dependon inhibition of mitochondrial oxygen consumption, which wouldallow delivered oxygen to pass through the peripheral mitochondrialzone to reach peroxisomes deep in the cell interior. We publishedrecently that exogenous NO induces bioluminescence in the intactfirefly; that NO mediates octopamine-induced bioluminescencein the dissected lantern, and that nitric oxide synthase isabundant in cells of the tracheolar system of the light organ.Additional experiments showed that nitric oxide gas (NO) inhibitsrespiration in isolated lantern mitochondria. Inhibition isreversed by bright light, and this inhibition is relieved whenthe light is turned off. Altogether, the results support theidea that NO triggers light production by reversible inhibitionof mitochondrial respiration in lantern cells, and probablyin tracheolar cells as well. The data also suggest that thelight of bioluminescence itself relieves NO inhibition thuscontributing to rapid on/off switching. While other mechanismsmay be in play, NO production that is directly related to neuralinput appears to have a key role in the oxygen gating that controlsflash communication signals.     

Retinal and optical adaptations for nocturnal vision in the halictid bee<Emphasis Type="Italic"> Megalopta genalis</Emphasis>

The apposition compound eye of a nocturnal bee, the halictid Megalopta genalis, is described for the first time. Compared to the compound eye of the worker honeybee Apis mellifera and the diurnal halictid bee Lasioglossum leucozonium, the eye of M. genalis shows specific retinal and optical adaptations for vision in dim light. The major anatomical adaptations within the eye of the nocturnal bee are (1) nearly twofold larger ommatidial facets and (2) a 4–5 times wider rhabdom diameter than found in the diurnal bees studied. Optically, the apposition eye of M. genalis is 27 times more sensitive to light than the eyes of the diurnal bees. This increased optical sensitivity represents a clear optical adaptation to low light intensities. Although this unique nocturnal apposition eye has a greatly improved ability to catch light, a 27-fold increase in sensitivity alone cannot account for nocturnal vision at light intensities that are 8 log units dimmer than during daytime. New evidence suggests that additional neuronal spatial summation within the first optic ganglion, the lamina, is involved.B.G. is thankful for travel awards from the Royal Physiographic Society, the Per Westlings Fond, the Foundation of Dagny and Eilert Ekvall and the Royal Swedish Academy of Sciences. E.J.W. is grateful for the support of a Smithsonian Short-Term Research Fellowship, the Swedish Research Council, the Crafoord Foundation, the Wenner-Gren Foundation and the Royal Physiographic Society of Lund for their ongoing support     

Studies on the seasonal biochemistry of the Northern krillMeganyctiphanes norvegica in the Kattegat

A well defined pelagic population of the Northern krill,Meganyctiphanes norvegica was sampled annually at a specific location in the Skandinavian Kattegat, and the major biochemical components were measured. Protein and lipid were the main constituents and underwent the most pronounced seasonal changes, clearly correlated to the prevailing supply of food organisms. The amount of lipid increased to a maximum of 48% of the dry weight towards winter, out of phase with gonad maturation, and therefore interpretable as deposition of overwintering reserves. Utilisation of stored reserves proceeds over winter, with loss of lipid, and decrease in weight but not in length. Comparison with literature data showed similarities with other krill populations from different geographical locations. The biochemical components ofEuphausia superba, the key organism of the marine Antarctic ecosystem, resembled those ofM. norvegica. Special polar adaptations are not obviously expressed in the proximate biochemical composition.     

The gas bladder of Argentina silus L., with special reference to the ultrastructure of the gas gland cells and the countercurrent vascular bundles

Summary The orifice between the two chambers of the gas bladder in Argentina silus is surrounded by a sphincter muscle. Gas analyses of the gas bladder contents of fish from 400 meters depth give 0–1% carbon dioxide and 9–72% oxygen. Micro-retia mirabilia form a countercurrent vascular system, and the arterial component has peripherally a sphincter mechanism. The function of the glandular layer of the anterior chamber remains uncertain, but the structure indicates secretion into blood capillaries. The lining epithelium of the anterior chamber may secrete some substance into blood or directly into the lumen, which may be involved in a secretory mechanism. This conclusion is not supported by our histochemical tests. The posterior chamber has no micro-retia and the blood vessels have a different origin from those of the anterior chamber. The blood vessels form a plexus of capillaries or sinuses in contact with the flat lining epithelium, thus allowing gases to pass freely by diffusion. — The muscular layers of both chambers are innervated by catecholamine-containing nerve fibres.The investigation was supported by grants from the Swedish Natural Research Council (No. 99-35) and by the Faculty of Mathematics and Science, University of Lund.     

Experiments on the physiology of southern and northern krill,Euphausia superba and Meganyctiphanes norvegica,with emphasis on moult and growth – a review

Physiological data are needed for life history studies on krill, and as parameters for input into energy budgets and models. In conjunction with moult and growth data, these may also prove useful for assessing the fishable biomass of krill. Here, the development of physiological concepts in experimental krill research is briefly evaluated, with emphasis on the gaps to be filled. Krill growth is very flexible, as well as strongly temperature and nutrition dependent. The polar Antarctic krill Euphausia superba grows as fast as the boreal species Meganyctiphanes norvegica, at least during the first 2.5 years, and the species are comparable in terms of physiological plasticity. Accordingly, as krill appear to adjust quickly to specific laboratory conditions, short-term experiments are essential if field conditions are to be reflected as closely as possible. Furthermore, direct comparisons between laboratory experiments and swarming studies in the field are advantageous. For these, M. norvegica is particularly well-suited, as swarms can be followed over longer times and more easily than in E. superba. For example, processes of moult and reproduction were found to be highly coordinated in swarms and populations of Northern krill. For this species a conceptual model of reproduction was developed based on a combination of short-term laboratory observations coupled with field data on moult and ovary stages. In further physiological experiments krill should be studied as groups when swarming. Using proxies, that is applying physiological and/or biochemical methods side by side, is a promising way to enhance the reliability of life history data.     

Moult cycle and seasonal activities of chitinolytic enzymes in the integument and digestive tract of the Antarctic krill,Euphausia superba

Summary Chitinolytic activity was quantified in euphausiid integuments in relation to moulting. In Euphausia superba, shortly before moult the activity increased in chitinase and N-acetyl--D-glucosaminidase to pronounced maxima indicating the onset of massive resorption of cuticular material. Enzymatic activity of E. superba corresponded to values in Meganyctiphanes norvegica, a boreal euphausiid which was investigated for comparison, as well as in insecta. Antarctic krill from winter catches displayed activities comparable to summer material suggesting physiological preparation for moulting. Accordingly, moulting did not cease during winter. Both enzymes were also active in the digestive tract in summer as well as in winter krill: chitin containing food of phyto-and zooplankton origin is digestable. Seasonally stable activities did not point to changes in nutritional preference. In contrast to other crustacea, digestive enzyme activity was not reduced around moult, suggesting a high capacity to continuously utilize food sources including chitin. This property can be linked directly to the high energy need caused by the necessity of constant active swimming in both krill species.Supported by German Research Counsil (DFG), grant-nos. Ad 24/9 and Bu 548/1Dedicated to Professor Dr. G. Hempel on the occasion of his 60th birthday     

Isoforms of an N-acetyl-β-d-glucosaminidase from the Antarctic krill, Euphausia superba: purification and antibody production

Two forms of the chitinolytic enzyme N-acetyl-β- -glucosaminidase (NAGase, EC 3.2.1.52) have been isolated from the Antarctic krill, Euphausia superba, in order to study their potential role in temperature adaptation processes. A chromatographic protocol was developed that allowed complete separation of the two enzyme forms, named NAGase B and NAGase C. The latter was purified to homogeneity with 600-fold enrichment and a yield of 17%. The molecular mass was 150 kDa. NAGase B showed characteristics of a glycoprotein due to affinity towards concanavalin A sepharose, while NAGase C did not. Highly specific polyclonal antibodies to NAGase C [anti-(E. superba-NAGase C)-IgG] showed only negligible cross-reactivity with NAGase B isoforms. A comparison with the Northern krill, Meganyctiphanes norvegica, revealed a corresponding chromatographic pattern with two main activity peaks, for differentiation named NAGase II and NAGase III. Application of the antibody on M. norvegica revealed a high specificity toward NAGase III and a low cross-reactivity with NAGase II. First indication is given that the two forms are no isoenzymes in a strict sense but instead may have different functions in the metabolism of krill.     

Lipolysis post mortem in North Atlantic krill

• 1.1. Three common species of North Atlantic krill, Meganyctiphanes norvegica (M. Sars), Thysanoessa inermis (Krøyer) and T. raschii (M. Sars), have been stored at 0°C post mortem, and the lipolytic activity followed by measuring changes in the lipid composition during storage.
• 2.2. Both phosphoglycerides and triacylglycerols were subjected to extensive hydrolysis with the formation of free fatty acids in all krill species examined, whereas wax esters, constituting a considerable proportion of the lipids in the Thysanoessa species, were not hydrolysed at all.
• 3.3. In M. norvegica the triacylglycerols and phosphoglycerides were hydrolysed at similar rates, whereas in T. inermis and T. raschii the phosphoglycerides were hydrolysed most rapidly.
• 4.4. For all krill species examined, the rate of production of free fatty acids was nearly constant during the initial phase of storage, and subsequently declined on prolonged storage.
• 5.5. At the end of the storage period of 16–24 days, the free fatty acids constituted about 35% of the total lipid in M. norvegica, and about 50% in the Thysanoessa species.
• 6.6. The rate of production of free fatty acids was about the same in all the three species of krill and seemed to be independent of the total lipid content.

     

Internal current generation in respiration chambers

A technical device generating a constant and directed current within a sealed respiration chamber is described. It does not involve any external pumps or tubing. This system is easy to handle, and improved the maintenance of rheotactic pelagic species like the Northern krill (Meganyctiphanes norvegica, Crustacea) or small fishes (Gasterosteus aculeatus) under experimental conditions.     

Abundance and distribution of early life stages of krill around Iceland during spring

Abundance, distribution and development of early life stages of krill (eggs, nauplii, calyptopes and furciliae) around Iceland were studied during the latter half of May 2013. Multivariate analyses were used to examine the relationships between water mass characteristics and phytoplankton spring bloom dynamics and distribution of krill. The results show that krill eggs, nauplii and calyptopes were most abundant over the shelf edges off the southwest and east coasts, while furciliae were most abundant on the shelf off the southwest coast. Meganyctiphanes norvegica and Thysanoessa longicaudata larvae were found mainly in the southwest, while T. inermis larvae were found in highest numbers on the east coast. Redundancy analysis showed that phytoplankton biomass, temperature and bottom depth explained 41% of the distribution pattern of early ontogenetic krill stages. In areas where krill eggs and larvae were most abundant (off the southwest coast), the phytoplankton spring bloom was in an advanced state, and the phytoplankton biomass and temperature were particularly high.     

Studies on metabolic properties in the Northern Krill, Meganyctiphanes norvegica (Crustacea, Euphausiacea): influence of nutrition and season on pyruvate kinase

The specific activity and the kinetic properties of partly purified pyruvate kinase (PK) (EC 2.7.1.40) from the Northern Krill, Meganyctiphanes norvegica, were investigated in relation to varying food resources. In order to evaluate the effect of starvation on the total energy metabolism, the respiration rates of fed and unfed krill were determined. The FPLC–elution profile of PK displayed two distinct peaks — PK I and II. The first isoform represented 80% of the total PK activity in the organism, and 20% was contributed by the second isoform. PK I was inhibited by ATP but was not influenced by fructose–1,6–bisphosphate (FBP). In contrast, PK II showed ATP inhibition and up to 2.5-fold increased activity by addition of 17 μmol·l⁻¹ FBP. The Michaelis–Menten constants of both isoforms were 2–10-fold higher for ADP than for phosphoenolpyruvate (PEP). Alanine showed no regulatory effect on PK I and II. In specimens starved for 7 days oxygen consumption decreased by 20%. Neither the feeding experiments nor the animals captured in the field during low and high productive seasons indicate that PK properties of M. norvegica are modified in relation to food supply. Accordingly, alternative mechanisms are involved in the depression of the metabolic rate in terms of oxygen consumption.     

Distribution and reproduction strategies of krill (Euphausiacea) on the Norwegian shelf

Meganyctiphanes norvegica and Thysanoessa inermis were the dominant krill species that were observed on the Norwegian shelf. T. inermis and M. norvegica were found in abundances of up to 14 and 22 no. m^?2, respectively, on the mid Norwegian shelf (Møre plateau, Halten, Sklinna, Træna banks) and up to the Lofoten area. The main distribution of the krill indicates that they are mostly restricted to the Atlantic waters of the shelf region in April and little krill was observed south of 62°N. The two dominant krill species appeared to have different strategies for the timing of spawning, thus avoiding direct overlap of spawning products. The majority of the T. inermis females were in the spawning condition in April. Meganyctiphanes norvegica appeared to spawn at least a month later. The sizes of the first time spawners of T. inermis were around 11–16 mm, and they were approximately 1 year old. The sizes of the second time spawners were around 17–22 mm. The spawning of krill on the Norwegian shelf coincides with the blooming of the spring phytoplankton in this region . Advection of eggs and larvae of krill, as well as other plankton, is important not only to the shelf and slope region, but also for the functioning of the wider Norwegian Sea ecosystem and the adjacent areas.     

Serotonin modulates a photic response in circadian locomotor rhythmicity of adults of the blow fly, Calliphora vicina

Abstract. The present experiments were undertaken to explore a role for serotonin (5-hydroxytryptamine, 5-HT) in modulating photic signal transduction in photoreceptors of the blow fly, Calliphora vicina. Injection of p-chlorophenylalanine (pCPA) into the haemolymph appeared to reduce sensitivity to the photic effects of constant ‘bright’ light (LL hyperactivity and circadian arrhythmicity). After drug injection in bright LL, flies continued with a free-running rhythm as in constant darkness (DD) or with a lengthened period τ as in ‘dim’ LL. When 5-HT was injected into flies kept in dim LL, they became hyperactive and arrhythmic as in bright LL. This finding suggests a potential role for serotonin as mediator of circadian changes in the insect visual system including extraretinal photoreceptors.     

Interannual variation in the diet of non-breeding male Antarctic fur seals, <Emphasis Type="Italic">Arctocephalus gazella</Emphasis>, at Isla 25 de Mayo/King George Island

The diet of non-breeding male Antarctic fur seals, Arctocephalus gazella, was investigated at Stranger Point, King George Island, through the analysis of scats during three consecutive summer seasons (1996, 1997, 1998). Overall, fish and krill were the most frequent prey occurring, respectively, in an average of 82.9% and 78.8% of samples (n = 131), followed by penguins (22.8%) and cephalopods (17.8%). Myctophids constituted almost 90% of the fish predated, with Electrona antarctica and Gymnoscopelus nicholsi being the most abundant and frequent species consumed. All fish taxa identified were krill feeding species suggesting that seals foraged mainly on a krill and a fish community associated with krill aggregations. However, a seasonal change was observed in the relative proportions of the different prey taxa, with a progressive decrease with time in the occurrence of krill and a concomitant increase of fish, penguins and squid. Possible influence of the strong 1997/98 ENSO event is discussed.     

The physiological response of Northern krill (Meganyctiphanes norvegica) to temperature gradients in the Kattegat

The Alkor-Deep (140 m), which forms part of a depression system in the northern Kattegat channel east of the island of Læsø (Denmark), is the location of a self sustaining population of Northern krill, Meganyctiphanes norvegica (Euphausiacea). This population is exposed to one of the most pronounced thermal gradients within the distributional range of this pelagic crustacean. During summer, the temperature of the water column ranges between 4 and 6 in the deep to 16 °C near the surface which results in the krill being exposed to temperature differences of 8–10 °C during diel vertical migration. Oxygen consumption rates were used to investigate the physiological adaptation of the animal to such gradients in temperature. The rates were found to increase exponentially from 31 mol O₂ h^-1 g_dw ^-1 at 4 °C to 72 mol O₂ h^-1 g_dw ^-1 at 16 °C, giving a Q ₁₀-value of 2.0, and indicating that physiological adaptation to varying thermal conditions does not take place. Behavioural adaptations are discussed which may help the krill to cope with large temperature gradients in their environment.     

<Emphasis Type="Italic">Rainfordia opercularis</Emphasis>, a liopropomin serranid (Teleostei: Serranidae: Epinephelinae): corroborative evidence from settlement-stage larvae

Rainfordia opercularis was described in 1923 from a single specimen taken in Edgecumbe Bay, Queensland, Australia. The species is rare in museum collections, and the larvae have not been described. In 1999, two settlement-stage larvae (20–21 mm in standard length) were collected in light traps set off Lizard Island, Great Barrier Reef, Queensland. The smaller of the two has one thin, flexible, extremely elongate dorsal-fin spine encased in a pigmented sheath. The larger specimen lacks an elongate dorsal-fin spine and exhibits caudal-fin pigment characteristic of adults. A combination of features in one or both of the settlement-stage larvae support the placement of Rainfordia in the epinepheline-serranid tribe Liopropomini: presence of an elongate, filamentous dorsal-fin spine serially associated with the first dorsal-fin pterygiophore; presence of a spine on the inner preopercular ridge; presence of dense pigment on the frontals; absence of an elongate spine at the angle of the preopercle; and absence of supraorbital spination. Supplementary material to this paper is available in electronic format at