Ultrastructure of the integument of a pelagic Crustacean: moult cycle related studies on the Antarctic krill,Euphausia superba

The ultrastructure of euphausiid integument was examined in relation to the moult cycle and supplemented by investigations of chitinase activity in the integument and content of N-acetyl--D-glucosamine in the hemolymph. The Antarctic krill, Euphausia superba was collected in 1983 in Admiralty Bay, King George Island, Antarctica. Some specimens of the Northern krill, Meganyctiphanes norvegica, from the Danish Kattegat served for comparison. As a major aim of the study, the moult staging system developed for living tissue could be verified by ultrastructural findings. Under experimental high production conditions of the Antarctic summer, no period of rest or intermoult between post- and premoult was observed in subadult E. superba. Neither was a resting phase seen at the cellular level, the epidermis remained active. The epidermal gland cells did not show any cyclical changes, and the organelles of protein synthesis were generally well developed in all moult stages. In order to follow the physiological course of events, structural and biochemical methods were combined and showed as a result that the last moult stage before ecdysis is characterized by massive cuticular resorption. The epicuticle remained ultrastructurally unchanged before and after ecdysis, even though its permeability should alter at ecdysis. The existence of muscle insertions which connect the old and the new cuticle across the exuvial space suggests an answer to the question why E. superba is hardly impaired in swimming almost up to the time of ecdysis.Dedicated to Professor Dr. H. Leonhardt on the occasion of his 70th birthday     

Ultrastructure and histochemistry of the integument of air-breathing polychaetes from mangrove swamps of sumatra

The integument of S.E. Asian air-breathing polychaetes exhibits a variable ultrastructure with regard to cuticle and epithelial cells. The cuticle varies in thickness, contents of regularly arranged collagen fibrils, and structure of the epicuticle. The basal portion of the epithelium is either of smooth outline, or consists of slender processes of the epidermal cells separated by wide intercellular spaces which are invaded by a loose system of glia cells and nerve fibres. Extracellular spaces within the epidermis of the gills of Dendronereides heteropoda are transformed into blood lacunae. The enzyme content of the epithelial cells is relatively low.Supported by the Deutsche Forschungsgemeinschaft (Sto 75/2, We 380/4).     

Symbiosis between bacteria and oceanic clonal sea star larvae in the western North Atlantic Ocean

Bacteria were present in the majority of clonal sea star bipinnariae (Luidia sp.) collected between June 1987 and August 1990 from oceanic populations in the subtropical western North Atlantic Ocean. Light and electron microscopy revealed dense aggregations of bacteria within lobes of the epidermal cuticle in the gastric region of larvae. Gram-negative rods and less-common elongate spiral bacteria were observed, always in close association with branching epidermal microvilli. Intact and partially digested bacteria occurred in vesicles and phagosomes within epidermal cells, a probable indication of phagocytotic activity by the host. The association of larvae of Luidia sp. and bacteria ranged widely on both geographical and temporal scales. During July and August 1988, larvae were abundant (1.6 to 11.1 m³) in the mixed surface layer (<10 to 100 m) of stations in the Gulf Stream, the Sargasso Sea and the North Equatorial Drift, and 96% (mean of 5 stations, n=8 to 10 larvae in each station) of the individuals surveyed harbored bacteria beneath their cuticle. In a single station on the Florida Current sampled during winter, spring, and summer months between 1989 and 1991, 79 to 90% of the larvae harbored bacteria. The incidence of symbiosis was higher in actively cloning larvae than in non-cloning larvae. This is the first documentation of a symbiosis in field populations of echinoderm larvae.     

Feeding of the copepod Acartia tonsa on the diatom Nitzschia closterium and brown algae (Fucus vesiculosus) detritus

Integumental sensilla were examined in 39 species of meso- and bathypelagic shrimps and 6 species of epibenthic shrimps. A tuft organ, consisting of a group of openended, tubular setae and so probably functioning as a chemosensor, is described. A single tuft with ancillary smaller tufts occur on the dorsal median surface of the fourth abdominal segment and two pairs of tufts occur on the proximal dorsal region of the telson of all oplophorid and pandalid shrimps examined. An additional tuft is present on the fifth abdominal segment of the rare bathypelagic Physetocaris microphthalma Chace. No tuft organs were identified in Processa canaliculata Leach, suggesting that they may not occur in all families of caridean shrimps. Acanthephyrid, systellaspid and pandalid shrimps have the integument completely covered by scales. These are extremely delicate and are normally removed during the process of catching the shrimp through abrasion of the integument by the net. The scales are peculiarly orientated, pointing anteriorly in the anterior half and posteriorly in the posterior half of the body. They probably have a sensory function as distance receptors, monitoring water currents and disturbances in the environment surrounding the shrimp.     

Anatomy of structures associated with air-breathing in Orchestia gammarellus (Crustacea: Amphipoda: Talitridae): coxal plates and gills

The structure of the coxal gills and coxal plates of the semi-terrestrial beachflea Orchestia gammarellus (Pallas) (Crustacea: Amphipoda: Talitridae) is described in relation to their possible use for aerial gas exchange and ion exchange. Anatomical evidence is presented to support the hypothesis that the medial surface of the coxal plates functions as an extrabranchial aerial gas-exchange site in O. gammarellus. Thus, the effective diffusion distance across the medial (or inside-facing) surface of O. gammarellus coxal plates (mean±SD=5.4±0.3 m; n=9, cuticle thickness 4.4±0.5 m, n=21) is only a third of the equivalent distance across both the coxal gills (18.4±6.0 m, n=10; cuticle thickness 1.7±0.6 m, n=7) and the lateral (or external) surface of the coxal plates (19.4±0.7 m, n=5; cuticle thickness 8.7±0.8 m, n=7). Chloride-ion-permeable areas were located using a silver-staining technique. All ten coxal gills appeared to be equally permeable to chloride ions after examination with a light microscope. However, the coxal plates and the rest of the integument do not appear to be chloride-permeable.     

Structural organisation,transfer and biological fate of endosymbiotic bacteria in gutless oligochaetes

From differences in size and structure, it appears there are two species of gram-negative bacteria in the gutless oligochaetes Phallodrilus leukodermatus and P. planus from Bermuda. A non-random, differentiated and consisten distribution pattern of the extracellular bacteria along the length of the worm's body underlines the regulated nature of the bacterial colonization. This emerges also from studies on the transfer of the bacteria between host generations: exceptional for oligochaetes, eggs are deposited singly in a sticky mucus sheath and not together in a cocoon. They become infected immediately at oviposition, apparently by intrusion from large stores of bacteria in a genital pad abutting the female pores. During ontogenesis, a balance is established between extracellular, active bacteria and intracellular lytic forms enclosed in vacuoles by the epidermal cells. In early developmental stages, lytic bacteria prevail, but older worms harbour mainly extracellular prokaryotes underneath their cuticle. The thick epidermis/cuticle complex is differentiated in regular zones with a progressive trend towards enclosing and digesting bacteria intracellularly in the deeper layers. These are the first results on the transfer and biological fate of endosymbiotic bacteria living in animals from sulfide biotopes.     

A preliminary approach to epidermal antimicrobial defense in the Delphinidae

In three delphinid species (Pacific spotted dolphin, Stenella attenuata; common dolphin, Delphinus delphis; harbor porpoise, Phocoena phocoena) the enzyme lysozyme and the peptide group -defensins are demonstrated for the first time in cetaceans as products of the thick integument. Lysozyme was found between the lamellae of the stratum corneum, in the cells of the stratum spinosum, free cells of the dermis, and in endothelial cells of dermal blood vessels. -defensins (especially type 3) were clearly located in cells of the upper stratum spinosum, and concentrated between the upper five or six layers of the stratum corneum. The occurrence and localization of such substances that may serve as a non-specific defense against bacteria, fungi, algae, and ectoparasites, was known until now only for haired marine mammals with typical skin glands, like in pinnipeds.Communicated by O. Kinne, Oldendorf/Luhe     

Comparison of gut morphology and gut microflora of seven species of mud shrimp (Crustacea: Decapoda: Thalassinidea)

The gut morphology is described for seven species of Thalassinidea; Callianassasubterranea (Montagu), Jaxea nocturna Nardo, Axius stirhynchus Leach, Calocaris macandreae Bell, Upogebia pusilla (Petagna), U. deltaura (Leach) and U. stellata (Montagu). Differences were observed in the structure of the dorsal and lateral teeth of the gastric mill. Callianassa subterranea and A. stirhynchus had simple robust dorsal teeth and proportionally larger lateral teeth with flexible spines. The three upogebiids had complex dorsal teeth that were highly ridged and proportionally smaller lateral teeth with inflexible spines. The dorsal and lateral teeth of J. nocturna were intermediate in form. The dorsal tooth of Calocaris macandreae, although relatively simple in design, possessed accessory projections, or “wings”, on either side of the main structure. The lateral teeth were proportionally quite large, but lacked ridges and spines. The epithelia surfaces of the digestive gland, midgut and posterior diverticulum were similar for all seven species. Differences were observed in the morphology of the hindgut. The hindgut lining was completely smooth for the three upogebiid species. The other thalassinidean species had four distinct rows of cuticular projections situated on top of the ridges formed by the longitudinal convolutions of the hindgut. The arrangement of these projections varied between species. A gut flora was observed for six of the seven species, the exception being C. macandreae. The differences observed in the structure of the gut are discussed in relation to feeding and thalassinidean phylogeny. Received: 3 July 1998 / Accepted: 8 October 1998     

An electron-microscope investigation into the cuticle and associated tissues of the operculum of some marine serpulids

The fine structure of the opercula of Spirobis borealis Daudin and S. pusilloides Bush, and the primary and secondary operculum of S. granulatus (L) have been examined by electron microscopy. In different regions, the epithelium of the operculum exhibits variations in thickness and fine structure which correspond to transitions from peduncular to opercular wall to opercular plate epithelia. Externally, the peduncle and walls of the operculum are covered by a cuticle which is composed of several layers of fibrils/fibres embedded in an amorphous matrix. It is regularly traversed by microvilli from underlying cells. At the operculum rim, the cuticle changes so that its appearance is thick and solid. It continues across the roof of the operculum as a thin layer overlying a calcareous plate. The secondary operculum of S. granulatus differs from those of the other species in that an internal calcareous plate and solid cuticle line the roof of the operculum. It differs further in possessing an opercular pore. The possibilities are discussed of the cuticle fibrils/fibres of the peduncle and the opercular wall being collagenous in nature, and later being transformed to a solid tanned structure overlying a calcareous plate. The modes of secretion of the cuticle and calcareous plate by underlying cells are also discussed.     

Spatial and temporal patterns of recuruitment of juvenile coral reef fishes to coral habitats within “One Tree Lagoon”, great barrier reef

Recruitment of juvenile fishes to coral grids, each comprising 4 colonies of 3 species of coral (Acropora formosa, Seriatopora hystrix and Pocillopora damicornis), was examined at 4 widely separated sites within the lagoon of One Tree Reef over two successive summers and intervening months (November 1976–January 1978). Recruitment was highly seasonal, with most recruitment occurring during summer. For many species, numbers settling differed greatly from year to year with total numbers (over all sites) differing as much as an order of magnitude between summers. Many fish species demonstrated marked preference among the three coral species as settling sites. The distribution of each of the 20 commonest species across the 4 lagoon sites differed significantly from a random pattern. Differences of an order of magnitude were common in the numbers of a given species recruited to different sites. Each site was preferred by at least one species. In each of 5 cases examined, the pattern of settlement of the species across the 4 sites changed significantly from one summer to the next. The distribution of recruits of a number of species corresponded to the distribution of adults, but for other species there was no correspondence. It is concluded that, at the spatial scale examined, patterns of recruitment of some taxa are consistently more variable than those of other taxa. The implications of spatial variability of recruitment for the distribution of adult fish is discussed and the importance of being able to discriminate such natural variability from other kinds of change is stressed.     

Functional feeding morphology of the euphausiid Nyctiphanes australis

The structure of the feeding basket, mandible and stomach armature of the krill Nyctiphanes australis (G. O. Sars, 1883) was examined by scanning electron microscope with the aim of predicting its diet. N. australis were collected during February 1986 and October 1986 to February 1987 in Otago Harbour, New Zealand (45° 50 S; 170° 37 E). Predictions based on the functional morphology were tested by examining stomach contents with SEM. Intersetule distances of the feeding basket (1 to 7.5 m) are finer than in other krill species, suggesting that N. australis can efficiently collect nanoplankton-sized particles (2 to 20 m). The mandibular edge index (0.74) matched the edge index of Meganyctiphanes norvegica. This indicates, in contrast to the fine feeding-basket setulation, that N. australis has a mandible resembling that of predominantly carnivorous krill species. The ratio of mandibular palp length to mandible width is 3.2±0.2, or relatively longer than the elongate palp thought to be indicative of herbivorous habit in Euphausia superba. The fine structure of the molar shows specialized surfaces which differ from those of other krill species. The internal armature of the stomach is heavily spinose, as is common in krill of herbivorous habit. Layers of various-shaped spines at differing densities were observed. The functional morphology suggests that N. australis is an opportunistic omnivore. The stomach contents tentatively support this prediction, containing fragments of phytoplankton and detrital material. However, stomach contents were generally amorphous, making identification of dietary components in the field difficult.     

Biochemical systematics of fishes of the genus Stegastes (Pomacentridae) from the Southern Marianas

The biochemical systematics of the subset of species of Stegastes (Pisces: Pomacentridae) available from coral reefs surrounding Guam, USA, was studied using three species of Pomacentrus as the outgroup. Analyses of allozymic variation at 14 presumptive loci (10 of which were polymorphic) using various evolutionary tree-construction techniques identified S. fasciolatus as the sister species of an assemblage comprised of S. albifasciatus, S. nigricans, and S. lividus: within this assemblage, S. lividus is the sister species of S. albifasciatus and S. nigricans, the pair of species which shared the most synapomorphic alleles and consistently exhibited the lowest degree of genetic distance relative to estimates for the other pairs of species studied. The consistency of ingroup topologies produced by parsimony analyses using Manhattan rectilinear distances computed from allele frequencies with those produced using the distance Wagner procedure and UPGMA (unweighted pair-group method of arithmetic averages) clustering, substantiates strongly the proposed phylogeny. The systematic scheme inferred from molecular data is concordant with predictions based on morphologic differences, i.e., differences in numbers of dorsal spines, that have been used as evidence in support of the proposal to partition Stegastes into at least two subgenera.     

“Glandular pockets” of the integument and feeding mechanism inPagurus bernhardus (Crustacea: Anomura)

The benthic anomuran crabPagurus bernhardus (L) were collected from the North Sea near Helgoland, West Germany, during the years 1986–1987. The histological study of the integument ofPagurus bernhardus reveals a double structure with dorsal and ventral cuticular layers and with two epithelial cell layers. Additional structures have been developed within the integument to trap micro food organisms. The structure and function of the cupshaped glandular pockets of the integument is described. The cells which secrete mucus are also described. Histological and histochemical methods have been employed to study the chemical nature of various cells of the integument and the secretions of the glandular pockets. The ecological significance of the sensory hairs, glandular pockets, cuticular layers and the supra-branchial groove is discussed with reference to the crab's feeding mechanism.     

Astaxanthin deposition in the cuticle of juvenile American lobster (<Emphasis Type="Italic">Homarus americanus</Emphasis>): implications for phenotypic and genotypic coloration

The phenotypic brownish-green color of an American lobster (Homarus americanus Milne-Edwards, 1837) is determined by genetic and dietary mechanisms. Diet influences color through the carotenoid astaxanthin. This pigment can appear red or blue depending if it is bound to protein, and the relative amounts of each form influence lobster color. Carotenoprotein formation was examined in sibling American lobsters rendered white by a diet low in astaxanthin, by feeding a diet containing 0, 55, 110, or 220 g astaxanthin g^–1 for 110 days beginning on 7 February 2004. The relative levels of red and blue in these lobsters were assessed through the analysis of digital photographs of the lobsters, which were taken monthly. The red/blue ratio was used to assess if free (red) or protein-bound astaxanthin (blue) was the dominant form in the lobsters, or if the two forms occurred in ratios allowing for natural coloration. Naturally colored lobsters developed only in the highest astaxanthin diet group, while lobsters fed the 55-g diet were blue. Differences were observed among different parts of the body, and were presumed to be a function of cuticle thickness. However, within each diet treatment, some lobsters initially became red, suggesting an accumulation of free astaxanthin, while other lobsters initially became blue, suggesting an accumulation of protein-bound astaxanthin. This variation may be a result of differences in the uptake of astaxanthin, the rate of carotenoid-protein complexing, or in the total amount of protein available for binding. This variability is likely to be the key to understanding the underlying basis of genetically determined color in American lobsters.Communicated by J.P. Grassle, New Brunswick     

Chitinolytic enzymes in the integument and midgut-gland of the shrimpPalaemon serratus during the moulting cycle

Chitinase andN-acetyl--D-glucosaminidase activity were quantified inPalaemon serratus (Pennant) integument and midgut gland during the moulting cycle. Studies were performed on specimens collected near Concarneau, France, in July 1989. The changes in specific activity are different in the two organs: in the midgut gland enzymatic activity is high throughout the whole moulting cycle with a weak peak at the early premoult Stage D1, whereas in the integument the activity of both enzymes is very low throughout post- and intermoult stages and rises only after premoult Stage D1. The highest specific activity is reached in D1 for chitinase and somewhat earlier (D1) forN-acetyl--D-glucosaminidase. The increase in specific chitinolytic activity coincides with an increase in ecdysteroids.     

Diets of some deep-sea brittle stars in the Rockall Trough

The stomachs of 1 165 individuals of the species Ophiacantha bidentata (Retzius), Ophiactis abyssicola (M. Sars), Ophiocten gracilis (O. Sars), Ophiura irrorata (Lyman), O. ljungmani (Lyman) and Ophiomusium lymani Wyville Thomson, collected from the general area of the Rockall Trough from 1973 to 1983, were examined; 47% contained material. This varied in nature and in volume between individuals, but overall, the diets of the 6 species showed quite high similarity. The few differences evident are interpreted as reflecting different lifestyles. Except for Ophiocten gracilis, the probably motile, epifaunal species (Ophiura irrorata, O. ljungmani and Ophiomusium lymani) showed the greatest variety in items, and the lowest similarity with the remaining species; they are probably unselective omnivores, eating whatever small prey or organic detritus they are able to find. Ophiacantha bidentata and Ophiactis abyssicola showed both the least dietary variety, and the highest similarity to each other. This probably reflects their more sedentary lifestyle, feeding on current-borne particles and small prey from perches on sessile fauna. The stomach contents of Ophiocten gracilis consisted mainly of amorphous organic material and mineral particles, indicating a microphagous feeding mechanism. With O. gracilis, the proportion of stomachs with food decreased markedly in summer, this coinciding with seasonal gonadal development when stomachs frequently appeared ruptured or compressed. The possible importance of seasonal sedimentation of fast-sinking particulates from the surface is discussed in relation to finds of flocculent organic material, often containing diatom frustules, in stomachs of all 6 species. Some doubts remain as to whether our results are biased by specimens egesting stomach contents on capture. As in shallow water, deep-sea ophiuroids seem to be trophic generalists lacking in dietary specialization. Brief notes on parasites encountered are also given.     

Distribution of the early-life phase of the Antarctic squid Galiteuthis glacialis in relation to the hydrology of the Southern Ocean in the sector 15° to 30°E

During R.R.S. Discovery Cruise 100 in the Southern Ocean (1979), 620 specimens of the Antarctic cranchiid squid Galiteuthis glacialis (Chun) were caught in the RMT8 opening-closing net, to the south of the Antarctic Polar Front (APF). The catch was dominated by individuals <20 mm dorsal mantle length (DML) and, apart from two specimens, the rest of the catch was <60 mm DML. Two larger squid (255 mm DML) were both immature and apparently female. The species is concentrated at a depth of 300 to 400 m; there is evidence of ontogenetic spreading and a shift in the modal depth of the population from the 300 to 400 m horizon in daylight to the 200 to 300 m horizon in darkness. The collection confirms the circumpolar distribution of the species and supports previous evidence that it rarely, if ever, occurs to the north of the APF. As in Mesonychoteuthis hamiltoni, the only other species of cranchiid squid known to occur south of the APF, the early-life phase of G. glacialis is concentrated in the upper zone of the warm deep water beneath the Antarctic surface layer where, putatively, there is a zone of enhanced biological activity.     

Spirorbinae (Serpulidae: Polychaeta) from South Africa,including three new species

Five Antarctic or south-temperate species of Spirorbiaae are centred in south-west Africa, whilst 6 tropical species extend round the south-east and another 4 are recorded from Mozambique. Of the three new species, Pileolaria (Pileolaria) annectans sp. n. has a rudimentary opercular plate surmounting a sort-walled sac of embryos, which somewhat resembles a thoracic brood pouch. Pileolaria (Pileolaria) dayi sp. n. has a helmet-like brood chamber, but this opens distally, like the brood cups found in the subgenus Duplicaria. Janua Pil-natalensis sp. n. has the dorsal collar folds usually separate, but partially fused in some juveniles.     

Ultrastructure of sperm and spermathecae in Micromaldane spp. (Polychaeta: Capitellida: Maldanidae)

Assessing the possibility that external fertilization has re-evolved requires the study of monophyletic groups that exhibit various reproductive methods. Maldanid polychaetes show a range of reproductive mechanisms, though previous studies of reproduction have hitherto been restricted to larger species with external fertilization. Micromaldane pamelae Rouse and M. nutricula Rouse are small, gonochoristic maldanids that brood directly developing larvae. Both species have sperm with elongate nuclei and an acrosome extending down each side of the anterior end of the nucleus. A true midpiece is absent; two mitochondria extend along the posterior region of the nucleus. Spermatids develop synchronously in large clusters connected by a cytophore. In M. pamelae sperm are released into the water as spermatozeugmata. These are comprised of clusters of sperm with their tails oriented to the centre and the sperm heads facing outwards. Females of M. pamelae and M. nutricula bear pairs of spermathecae ventrally (M. pamelae three pairs, between setigers 10 and 11, 11 and 12 and 12 and 13 and M. nutricula two pairs, between setigers 10 and 11 and setigers 11 and 12). The blind sacs are epidermal invaginations bound closely together. The entrance to each spermatheca may only be 1 to 2 m across with each spermatheca holding several hundred sperm. This represents the first detailed study of spermathecae in the Capitellida. The occurrence and structure of spermathecae and spermatozeugmata in other groups are discussed and compared with Micromaldane spp. Comparisons are made with non-polychaetes with the purpose of discussing functional aspects of reproductive mechanisms in marine metazoans in general. Elongate sperm nuclei are associated with sperm storage and/or large egg size. The lack of an elongate sperm midpiece may be an indicator of having to swim in water but does not contraindicate sperm storage. Spermatozeugmata may serve as an indication of sperm storage and brooding of larvae. Speculations on the phylogenetic significance of these reproductive features are limited by the fact that supposedly modified (i.e., derived) states may reflect functional/structura, constraints of small body size.     

Effects of zinc on epidermal ultrastructure in the larva of Clupea harengus

In March, 1983 changes in epidermal ultrastructure were examined in Clupea harengus L. larvae hatched from eggs incubated in four zinc concentrations (0.5 2.0, 6.0 and 12.0 ppm). In addition to the outer and inner epidermal cell types described previously, a third type of cell is present. Ultrastructurally this resembles the epidermal chloride cells of Sardinops caerulea Girard, characterised by numerous mitochondria, extensive smooth endoplasmic reticulum and a free surface exposed to the environment. These cells occur on the yolk sac, head and in the regions of the trunk just above the yolk sac. In larvae treated in 12.0 ppm zinc, the cells contain fewer mitochondria with fewer cristae than those in the controls. Smooth endoplasmic reticulum is much reduced forming nodular masses and contains granular osmiophilic inclusions. In larvae hatching from eggs previously incubated in 6.0 and 12.0 ppm zinc, the epidermal cells contain more vesicles, intracellular spaces, and swollen mitochondria and show signs of necrosis. The viability of these larvae is discussed.