Study of surface-shell features in thecosomata (Pteropoda: Mollusca) by means of scanning electron microscopy

The shell ornamentation of 6 species of Limacinidae and 2 species of Peraclididae has been studied by means of scanning electron microscopy. The author analyzes the phylogenetic relationships between the species considered and the morphologic resemblance with protoconchs of other Gastropoda. The function of some of the features observed is considered, and the relationship between adult and larval environments and shell formation discussed. Pteropoda do not exhibit conspicuous differences between veliger and adult shells, but there is quite an important difference between the protoconch and the rest of the shell, the former being very similar in several Gastropoda. The division of the genus Limacina into 3 subgenera, as proposed by Van der Spoel (1967), is also partially confirmed by the present study.     

Particulate pollution case studies which illustrate uses of individual particle analysis by scanning electron microscopy

Individual particle analysis by scanning electron microscopy is a high resolution analytical technique which can provide the detail necessary to solve many particulate pollution problems. In an electron microscope a beam of electrons is focused on a specimen resulting in a number of signals which are used to collect chemical and morphological information from individual features either manually or automatically. Case studies are presented which illustrate the applications of the technique to three paniculate pollution problems of differing complexity.     

Are barnacle larvae able to escape from the threat of UV?

Earlier laboratory experiments suggested that environmental levels of UV-B radiation can damage the eyes of barnacle naupliar larvae and impair their phototactic behaviors. However, since barnacle larvae may avoid UV by migrating to deeper waters, it is not known whether such impairment would actually occur under field conditions. For the first time, this study provides both field and laboratory evidences to show that prevailing UV-B in the natural habitat of barnacle larvae could be an important environmental factor affecting natural barnacle populations. We here showed that although barnacle nauplii may avoid UV-B irradiation by downward migration, the amount of UV energy (9.8 × 10⁻⁶ J) received by a naupliar eye during downward migration in the natural water column is within the same order of magnitude as the total energy (7.5 × 10⁻⁶ J) sufficient to cause damages to naupliar eye and impair their phototactic responses. It is possible that solar UV-B prevailing at shallow waters would pose a similar threat to other zooplankton species over large geographic scale.     

The surface attachment structure: a unique type of integumental formation in neustonic copepods

A unique type of integumental formation is described for several members of the copepod family Pontellidae. This surface attachment structure (SAS) consists of a mass of fine setules arranged in two semicircles on a flattened area of the anterodorsal surface of the cephalosome. Using transmission electron microscopy, the SAS was shown to be continous with the cuticle and not linked to chemo- or mechanosensory cells; its function is purely mechanical. This structure is probably an energy-saving means for these large and heavy neustonts to stay attached to the surface film. The SAS is species-specific and may thus be of potential importance to the systematics and phylogeny of the Pontellidae, in the same manner as integumental pores and sensilla, which form patterns characteristic of several copepod families and genera.     

Influence of substratum heterogeneity and settled barnacle density on the settlement of cypris larvae

On the Atlantic coast of Canada, Semibalanus balanoides (L.) is widely distributed in the mid-intertidal zone, whereas in the Gulf of St. Lawrence, this species is mostly limited to crevices. We tested the hypothesis of regional differences in microhabitat selection by barnacle larvae at settlement in 1984 and 1985 at St. Andrews, New Brunswick, Canada. Since larvae settle in microhabitats already colonized by adults, the relative influence of settled barnacle density and of different scales of substratum heterogeneity on settlement were evaluated experimentally at Capucins, Québec, (Gulf of St. Lawrence) and at St. Andrews, New Brunswick (Atlantic coast). On a large scale (>10 cm deep crevices) of heterogeneity, results show that, in the Gulf, cypris larvae settled nearly exclusively (93%) in natural crevices rather than on adjacent horizontal surfaces. On the Atlantic coast, settlement was more important outside than inside of crevices, when the substrata were either natural or artificial. This result is unique and contrasts sharply with all known reports on barnacle settlement in relation to surface contour. The influence of barnacle density on settlement was greater than that of large scale heterogeneity. On a small scale (<1.5 cm deep cracks), the presence of conspecifics had a stronger effect on settlement than heterogeneity in both regions. Field observations showed a relationship between larval settlement density and percentage of adult cover. Settlement increased up to 22 or 30% (Gulf and Atlantic coast) of adult cover and decreased afterwards. The results confirm the hypothesis of larval selection for cryptic habitats in the Gulf and the opposite behaviour (preferences for horizontal surfaces) on the Atlantic coast. This microhabitat selection is apparent at large scales of heterogeneity, whereas at small scales, the presence of conspecifics is the predominant factor.Contribution to the programme of GIROQ (Groupe interuniversitaire de recherches océanographiques du Québec)     

Morphology of grooming limbs in species of Petrolisthes and Pachycheles (Crustacea: Decapoda: Anomura: Porcellanidae): a scanning electron microscopy study

The morphology of the grooming limbs, the 5th pair of pereiopods, was studied by scanning electron microscopy in six species of porcellanid crabs, Petrolisthes cabrilloi, P. cinctipes, P. armatus, P. galathinus, Pachycheles monilifer, and Pachycheles rudis, and their function was inferred by comparison with findings from previous studies. Grooming limb morphology was almost identical among the four Petrolisthes species and differed little compared to that of the two Pachycheles species. The 5th pereiopods bore a basal tuft of mechanoreceptive setae, three different types of grooming setae armed with setules or denticles, two types of smooth sensilla for location and identification of fouling objects, and a terminal, toothed chela for picking firmly attached objects off the gills and body. The grooming limb was extremely flexible and could reach most parts of the body, including the gill chamber on the opposite side. The grooming limb morphology in Petrolisthes cabrilloi is consistent with its wellknown effectiveness in preventing parasitism by the rhizocephalan Lernaeodiscus porcellanae. Grooming setae remove recently attached cyprids, while the chela can grip and remove the much smaller, firmly attached kentrogons. Porcellanid crabs not known to host rhizocephalans, however, had grooming limbs almost identical to those of Petrolisthes cabrilloi despite their previously demonstrated failure to prevent settlement and infestation by L. porcellanae larvae. The effectiveness of P. cabrilloi in removing kentrogons, therefore, seems also to depend on behavioral adaptations whereby this species recognizes the parasite larvae as high-threat objects.     

Testicular cellular toxicity of cadmium : transmission electron microscopy examination

It is clear that environmental heavy metals influence life systems and reproductive system. In the present study histological investigation revealed that cadmium was testicular toxicant in mice. Here we compared the fine-structure of spermatogenesis in two groups of mice (SWR), experimental and control. The experimental group underwent cadmium ingestion at 1 mg/kg daily for 4 weeks. The control group underwent ingestion of distilled water with equal dosages, using the same type of injectors, for 4-weeks. After cadmium exposure period both control and experimental groups were killed and samples of the testes were processed for microscopic examination. Ultra sections were examined and photographed by Transmission Electron Microscope (JEOL- 100SX) at 80KV. Ultrastructure examination revealed, vascular endothelial, interstitial, and sertoli cells damages. Early impairments of germinal cellular differentiation resulted in deformations in all parts of late spermatid. There were dislocation of accrosomal granules, nuclear damage associated with chromatin heterogeneity, detached spermatid from the apical process of sertoli cell, disarrangement of the mitochondria, abnormal oriented tail piece, and abnormal microtubules complex. These ultra morphological abnormalities relate to cell injury and to the resulting physiological abnormality, necrobiosis. Based on the results of this investigation it can be concluded that cadmium ingestion at 1000 microg/kg caused testicular toxicity and abnormalities in early sperm development.     

Analysis by slow-motion picture photography and scanning electron microscopy of radular function in Urosalpinx cinerea follyensis (Muricidae,Gastropoda) during shell penetration

Radular function in the muricid gastropod Urosalpinx cinerea follyensis Baker during shell penetration was examined with slow-motion picture photography and scanning electron microscopy. Particular attention was paid to possible injury of buccal structures by radular cusps. The presence of a flexible cuticulated buccal armature, and delicate synchronization of movements of odontophoral cartilages, subradular membrane, teeth, and buccal mass, explain the absence of shredding of live buccal tissues. Some light abrasion was evident, but generally only in the cuticulated gully of the subodontophoral shield, on the rim of the jaw, and on the anterior edge of the esophageal valve. Rasping at the surface of incomplete bore-holes is done by major cusps of rachidian teeth over the bending plane. Marginal teeth lie on the slopes of the odontophore, generally remain depressed below the level of rachidian teeth, and thus scrape the shell only lightly, if at all. The sharp posteriorly recurved shape of central and lateral rachidian cusps enhances their scooping effectiveness. These teeth produce smooth, conspicuous traces in the soft shell of Mya arenaria and shallow traces in the harder shell of Mytilus edulis Linné. The impact of individual cusp strikes was not evident in traces. With wear, rachidian cusps become increasingly blunted, a reflection of their ploughing action over chemically weakened shell, and are eventually sanded flat. Marginal teeth wear primarily at their ends, the tips becoming truncated as they pass lightly over the shell surface. The advancing edge of the odontophore during rasping strokes, plotted on the image from motion pictures moves slowly at first, then more rapidly in the middle of the stroke, and slows again at the end. Duration of strokes ranged from 0.45 to 0.75 sec. Duration of rasping cycles varied from 1.3 to 2.0 sec. The number of transverse rows of teeth passing over the bending plane during the rasping stroke varied from 14 to 32, and the average time for the passage of one transverse row over the bending plane ranged from 20 to 48 msec. The number of transverse rows of teeth remaining exposed beneath the odontophore below the bending plane at the end of the rasping stroke, and between the bending plane and the anterior end of the sulcus in the radular diverticulum, was approximately 10.     

Impact of diesel oil effluent in the mucosal surface of the alimentary canal of Oreochromis nilotica (Linnaeus): a scanning electron microscopic study

Adverse effects of diesel oil on microanatomical structure of the alimentary canal of O. nilotica were studied using SEM observations. The study revealed irregular arrangement of the stratified epithelial cells along with fragmentation of the normal concentric pattem of microridges of the same cells in buccopharynx and oesophagus. The excessive secretion of mucus of buccopharynx and oesophagus were the salient changes caused by diesel oil pollution. The destruction and degeneration of the mucosal folds of stomach and intestine along with their epithelial cells exhibited a concrete hyperactivity resulting in abundant' secretion of mucus over the microridges of the epithelial cells. In the intestinal region the columnar epithelial cells showed tumefaction and microvilli of the plasma membrane of epithelial cells get heavily damaged. Disarray of the microridges of epithelial cells, excessive secretions of mucus formation of even cell sheet were the most conspicuous changes in rectum. It was concluded that chronic exposure of diesel oil may hamper the absorption of the nutrients through alimentary canal resulting into ill-growth and production of the fish.     

Settlement behavior and antennulary biomechanics in cypris larvae of Balanus amphitrite (Crustacea: Thecostraca: Cirripedia)

The settlement behavior, antennulary biomechanics and morphology were investigated in the intertidal cypris larvae Balanus amphitrite. The very maneuverable antennule of the cyprid consists of four jointed segments, and it is the prime appendage for substratum exploration and final settlement. We recorded several previously undescribed settling behaviors, such as the "tight direction change", for this intertidal species and discovered that the antennulary morphology is modified to facilitate such behaviors. Segment 2 has a modified lateral cuticle, which gives the segment an additional higher degree of maneuverability compared to non-intertidal cirripede species of Rhizocephala. Additional supporting muscle groups were also found in the antennule of our species, which were not found in the investigated rhizocephalan larvae. The results could very well be associated with the higher needs of substratum recognition in intertidal species, due to more heterogeneous habitats such as intertidal rock flats.     

Inhibition of attachment of larval barnacles,Balanus amphitrite,by bacterial surface films

Films of bacteria on solid substrata can positively or negatively influence the attachment of marine invertebrate larvae. Effects of marine bacteria on the attachment of cypris larvae of the barnacle Balanus amphitrite Darwin were examined in the laboratory. Bacteria, grown to mid-exponential phase and allowed to adsorb irreversibly to polystyrene petri dishes, attached in densities of 10⁷ cells cm^-2. Assays (22h) were used to compare the effects of adsorbed cells of 18 different bacterial species on larval barnacle attachment. Most of the adsorbed bacteria either inhibited or had no effect on larval attachment compared to clean surfaces. Experiments testing the effect of larval age on barnacle attachment were conducted with six species of bacteria and showed that older larvae attached in higher percentages to clean surfaces and that bacterial films generally inhibited larval attaschment. Both the species of bacteria and the in situ age of the adsorbed bacteria affected barnacle attachment response: older films of Deleya (Pseudomonas) marina were more inhibitory. Bacterial extracellular materials may be involved in the inhibitory process.     

Antifouling adaptations of marine shrimp (Crustacea: Decapoda: Caridea): Functional morphology and adaptive significance of antennular preening by the third maxillipeds

Preening (cleaning, grooming) of the antennules and other cephalothoracic appendages by the third maxillipeds was observed in several species of shrimp. Distribution and ultrastructure of serrate grooming setae on the third maxillipeds, which scrape the antennules, was studied with light and scanning electron microscopy. The motor patterns of antennular cleaning were similar for all species. Antennular preening was the most frequent grooming behavior observed, but the duration of other grooming behaviors was greater. Tidepool shrimps (Heptacarpus pictus) experimentally prevented from grooming the antennules by ablation suffered fouling of the olfactory hairs of the antennules with their subsequent breakage and loss; antennules of controls remained clean and undamaged. Antennular preening, a frequent and widespread behavior of caridean shrimps and other decapod crustaceans, is suggested as having high adaptive value in keeping sensory sites free of epizoic and sedimentary fouling which might render them inoperative.     

Development of the pericalymma larva of Solemya reidi (Bivalvia: Cryptodonta: Solemyidae) as revealed by light and electron microscopy

Solemya reidi Bernard 1980 is a gutless protobranch bivalve known to possess intracellular chemoautotrophic bacterial symbionts in its gill. A light and electron microscope study on the embryology and larval development of S. reidi provides data for the bivalve Subclass Cryptodonta. S. reidi spontaneously spawned large eggs (271 m in diameter), which developed within individual gelatious egg capsules. The first several cleavages were equal and a distinct molluscan cross was formed at the animal pole of the embryo, features previously unreported in bivalve development. Lecithotrophic pericalymma larvae (similar to the larvae of paleotaxodont protobranch bivalves and aplacophoran molluscs) hatched at 18 to 24 h and remained in the water column for a further 5 d at 10°C. At hatching, larvae measured from 360 to 440 m in length and from 225 to 265 m in cross-sectional diameter. Definitive adult structures developed within an epithelial locomotory test entirely covered with compound cilia. The test histolysed at metamorphosis and was ingested throught the mouth into the perivisceral cavity. Length and height of the shell following metamorphosis was 433 m (±42 m, n=16) and 282 m (± 29 m, n=13), respectively. Primary data and data from the literature show that the type of larval development in both paleotaxodont and cryptodont bivalves cannot be reliably estimated from egg or prodissoconch sizes.     

A scanning electron microscope study of the mandibular morphology of boreal copepods

Morphological details of the mandibular blades of 11 species of copepods were examined with scanning electron microscopy. The micrographs improved our understanding of the complex toothed edge of the mandibular blade, which in turn aids in identification of the copepod prey of chaetognaths by gut-content analysis. Beklemishev's discovery of siliceous tooth crowns in calanoid copepods was confirmed for most of the species examined. Long, sharp projections on the crowns of herbivorous species, and the deep grooves into which they fit on the teeth of the opposite mandible, suggest a cracking rather than a grinding function for these teeth.     

Prey-predator dynamics in rotifers: density-dependent consequences of spatial heterogeneity due to surface attachment

Classical models of prey-predator interactions assume that per capita prey consumption is dependent on prey density alone and that prey consumption (functional response) and consumer proliferation (numerical response) operate on the same timescales and without time lags. Several modifications have been proposed for resolving this timescale discrepancy, including variants where the functional response depends on both prey and predator densities. A microcosm system with the rotifer Brachionus 'Nevada' feeding on the prasinophyte Tetraselmis sp. showed significant (P < 0.0005) increases in steady-state biomasses of both prey and predators with increasing carrying capacity (represented by total phosphorus of the growth medium), which is inconsistent with predictions based on the traditional prey-only-dependent functional response. We provide data indicating that surfaces where the predator can attach provide a high-quality habitat for rotifers, which can result in a predator-dependent functional response. We also show that partitioning between the attached and free-swimming habitats was fast compared to the timescale of the numerical response. When attached to surfaces, rotifers maximized net energy gain by avoiding the high cost of swimming and by increased food capture due to reduced viscous drag. A mathematical model with prey-dependent functional response and wall-attached and free-swimming fractions of the population describes our data adequately. We discuss the implications of this finding for extrapolating microcosm experiments to systems with other surface-to-volume ratios, and to what extent our findings may apply to other popular model organisms for prey-predator interaction.     

Epifluorescence microscopy,a technique for the study of feeding in Crassostrea virginica veliger larvae

Epifluorescence microscopy was applied as a new technique to observe the response of Crassostrea virginica veliger larvae to the algal foods Monochrysis lutheri or Chlorella autotrophica. Uptake, lysis, and digestion or rejection of these algae by oyster larvae were observed microscopically through the autofluorescence of chlorophyll a and its derivatives. Both species of algae were taken up by large numbers of larvae within 5 min after being added to larval cultures. The good growth of larvae obtained with M. lutheri as a food source was related to rapid uptake, lysis, and digestion of this alga. Larvae fed with C. autotrophica did not grow. Althogh C. autotrophica was also taken up rapidly by the larvae, there was no lysis or digestion of these algae and cells were eventually voided.     

Scanning electron microscopy investigation of bacterial colonization of the marine copepod Acartia clausi

Association between copepods and bacteria was observed in many scanning electron micrographs. Particular sites on the copepods were selectively colonized by bacteria; the joints of segments and legs, swimming legs and depressed parts of the body surface were found to be densely covered with bacteria. In comparison, bacterial attachment to copepod skeletons in fecal pellets excreted by chaetognaths was not selective; bacteria were sparsely found all over the copepod. Between 9 to 30% of copepods in Tokyo Bay waters had attached bacteria in January and April 1983.     

Surface morphology of dogfish (Scyliorhinus canicula) gill epithelium,and surface morphological changes following treatment with zinc sulphate: A scanning electron microscope study

The surface morphology of the gill epithelium of the dogfish Scyliorhinus canicula L. (collected near Barcelona, Spain, in February–March, 1981) was studied by scanning electron microscopy. Pavement cells exhibited either surface microvilli or microridges, which were randomly distributed on both the primary (afferent and efferent sides and interlamellar spaces) and secondary epithelium. Chloride cell apical regions on the afferent side displayed characteristics closer to freshwater than to marine teleosts: no apical pits were detected; chloride cells displayed longer microvilli than those of adjacent cells. Two morphologically different cell types were identified: a large chloride cell and a smaller cell (probably a chloride cell too), measuring 4 to 7 m and 1 m, respectively, the latter being dominant in the interlamellar spaces. Apart from pavement cells, the mucous cell was the prevalent cell type on the efferent region. The respiratory epithelium consisted of a mozaic of typical epithelial cells; some chloride and mucous cells were present, mainly located at the base of the secondary lamellae. Surface morphological changes were monitored after exposing the dogfish to subacute zinc treatment: 10 ppm Zn (ZnSO₄) for 3 wk. The chloride cell was the only cell type that underwent any modifications: microvilli became longer and tips were swollen following Zn treatment. The results are discussed in relation to a previous study on the effects of zinc sulphate on chloride cell response and heavy metal distribution in excretory organs of the dogfish.     

Phytoplankton composition and organic matter organization of mucous aggregates by means of light and cryo-scanning electron microscopy

Two types of northern Adriatic mucous macroaggregates—surface and cloud-like—collected in June 2000 at the sea surface and at the depth of the pycnocline, respectively, were studied by light and cryo-scanning electron microscopy. The combination of both techniques permitted investigation of species/particle composition and particle–organic matrix associations of copious mucous aggregates. Both types of macroaggregates had, on average, heterogeneous composition with diatoms the dominant biotic component, differing by the prevailing species and stage of degradation. Some differences in the organization of the basic fibrillar organic network of macroaggregates were observed. Clouds were characterized by a tighter organization of the polysaccharidic fibrils within the walls of the organic network constituting the cloud. This organization was looser in the organic network within surface aggregates. Plankton remains and inorganic particles could lead to lower porosity and greater rigidity of these aggregates. The more complex organization of the organic matter and the intense growth of Cylindrotheca closterium within the clouds probably reflect a longer physical diagenesis of the polysaccharidic fibrils in comparison to surface aggregates.Communicated by O. Kinne, Oldendorf/Luhe