Anatomy and ultrastructure of the marine oligochaete Tubificoides benedii (Tubificidae), with emphasis on its epidermis-cuticle-complex

Although Tubificoides benedii (d'Udekem, 1855) (= Peloscolex benedeni) is a ubiquitous form in eutrophicated or polluted coastal muds and is characterized by an exceptional viability in sulphide sediments, almost nothing is known about its anatomy and the structure of its conspicuously papillate body surface. As a part of a research project on sulphide annelids, done by our group, we investigated the body structure of this common and still extraordinary marine tubificid using light, scanning and transmission electron microscopy.While the internal structures correspond to the general pattern of marine tubificids (Giere, 1983), the epidermis-cuticle-complex is unusual. The epidermis cells, which are much interdigitated, contain numerous extremely long and irregularly shaped mitochondria. At the tip of the body, many sensory cells, embedded in the normal epidermis cell layer, end with ciliary tufts at the body surface. Long epidermal microvilli traverse the thick cuticular layer which only in the anterior- and posterior-most segments is studded by epicuticular projections. From most parts of the body these surficial rodlets, so far held typical for all Oligochaeta, are absent. Instead, here the cuticle forms numerous high, almost leaf-life papillae between which a viscous mucus layer regularly harbours many bacteria.This distinct papillate body armature may serve for stabilization of the mucus layer with its associated bacteria. It is well conceivable that the mucus, beside its usual role of reducting friction in the burrowing process, attracts bacteria. That their settlement may be more than an accidental association and involve some regulative interaction is indicated by the specific attachment of gram-negative bacterial threads often populating the posterior end of T. benedii (Dubilier, 1986). The significance of possible stress exerted by the sulfide-environment on the worm, as evidenced by an unusually high concentration of lysosomal structures and abnormally formed mitochondria in the epidermis, has to be verified in further studies.     

Identity and ecophysiology of filamentous bacteria in activated sludge

Excessive growth of filamentous bacteria in activated sludge wastewater treatment plants (WWTPs) can cause serious operational problems. With some filaments there may be the problem of bulking, where inadequate flocculation and settling of the biomass in the secondary clarifier results in a carryover of solids with the final treated liquid effluent. Their proliferation often encourages the development of stable foams on the surface of the reactors, and these foams may impact negatively on plant performance and operation. The availability of culture-independent molecular methods now allows us to identify many of the more common filamentous organisms encountered in WWTPs, which are phylogenetically diverse, affiliating to seven separate bacterial phyla. Furthermore, the extensive data published in the past decade on their in situ behaviour from the application of these culture-independent methods have not been summarized or reviewed critically. Hence, here, we attempt to discuss what we now know about their identity, ecophysiology and ecological niches and its practical value in better managing activated sludge processes. Some of this knowledge is already being applied to control and manage full-scale WWTPs better, and the hope is that this review will contribute towards further developments in this field of environmental microbiology.     

Morphological and Ecophysiological Adaptations of the Marine Oligochaete Tubificoides benedii to Sulfidic Sediments

SYNOPSIS. The marine oligochaete worm Tubificoides benedii inhabitscoastal tidal sediments in which sulfide can reach toxic concentrations.The role of external ironsulfide deposition in sulfide detoxificationis discussed together with a review of morphological and ecophysiologicaladaptations of T. benedii to sulfide. The body wall of T. benediiturns black in the presence of sulfide. Histochemical studiesand micro-X-rayanalyses provide evidence for the reaction ofiron in the mucus layer above the cuticle of the worm with environmentalsulfide to produce ironsulfide. The deposited ironsulfides areeither reoxidized or shed off through moulting, a process otherwiseunknown in oligochaetes. However, calculations on the diffusionrate of sulfide into T. benedii show that the deposition ofironsulfides do not play an important role in sulfide detoxification.The first and last few segments of T. benedii are not blackenedby sulfide and do not appear to precipitate sulfide. The diffusionrate of sulfide through these segments is so rapid that internalsulfide concentrations reach levels inhibitory to cytochromec oxidase, the key enzyme of aerobic respiration, within minutes.When internal sulfide concentrations increase to toxic levels,reliance on an anaerobic metabolism represents a successfulmechanism of sulfide tolerance in T. benedii. Metabolic adaptationsto hypoxia and sulfide include the maintenance of aerobic pathwaysdespite low oxygen or high sulfide concentrations and the abilityto gain energy through anaerobic pathways when oxygen and/orsulfide concentrations become limiting     

Some ultrastructural observations on the integument of a pentastomid

The cuticle of the cephalobaenid pentastomid Reighardia sternae is described at various stages of the moult-intermoult cycle. The intermoult cuticle comprises four layers: an outer epicuticle; an underlying dense layer, the protein epicuticle; a fibrillar endocuticle; and a denser subcuticle. The overall similarity between the structure and composition of these layers and those of insects is discussed. However, the orientation of the chitin-protein fibres in the endocuticle does not show the rotating structure characteristic of many arthropod species, but this does appear in the sclerotized hooks. It is suggested that this comparatively loose, poorly oriented endocuticular structure produces a highly extensible cuticle which is precisely adapted to the specialized, endoparasitic habit of this species. Events at ecdysis, particularly the secretion of moulting fluid and the deposition of cuticulin, follow the insect pattern precisely. The phyletic significance of these observations is discussed.     

Some ultrastructural observations on the microfilaria of Breinlia sergenti--nervous system

The amphids, cephalic papillae, phasmids, excretory complex, anal vesicle, rectal cells, G cell and the nerve ring are described in the microfilaria of Breinlia sergenti. The significance of these various structures in playing a nervous role in this organism is discussed, and a working hypothesis presented to explain the nervous co-ordination in this parasitic stage.     

Some ultrastructural aspects of spermatogenesis inLycopodium complanatum

Summary Analysis of thin sections shows that the blepharoplast in an early spermatid ofLycopodium consists of two basal bodies and the subtending spline apparatus. The latter has a 4-layered or Vierergruppe organization much like that reported for certain bryophytes and vascular plants. During the course of spermatid maturation the vertically lamellate S₂ stratum of the spline apparatus is transformed into a thinner layer of dense osmiophilic material, although the vertical lamellae of the S_3,4 strata persist. Rarely there occurred double Vierergruppen, presumably anomalous structures in which a single layer of spline microtubules is flanked on each side by two separate sets of S_2–4 strata. Longitudinal sections show that the basal bodies in an early spermatid lie parallel and close together with their microtubular triplets imbricating in the same direction. In late spermatids these basal bodies lie antiparallel and widely separated, their triplets now imbricating in opposite directions. This change is intrepreted to result from the relocation of one of the basal bodies to a position halfway around the cell by moving distal end first over the subtending spline. The basal bodies become invested by a globular-textured material not previously observed in plant spermatids. The role of this investing material is tentatively thought to be related to anchorage of the basal bodies.     

Mysterious `crystals': found attached to the epidermal peritoneum of marine tubificid (Annelida,Clitellata) species

Marine tubificids are abundant and diverse in the carbonate sediments of Bermuda, as well as in many other tropical and subtropical locations. Recently, during microscopic observations of living specimens, crystal-like structures were observed attached to the coelomic peritoneum and in the coelomic cavity of some Bermuda species, including phallodrilines of the genera Aktedrilus and Pectinodrilus,and a rhyacodriline of the genus Heterodrilus. Similar structures were not seen in tubificid species of Thallasodrilides and other limnodriloidines, a second species of Heterodrilus, a tubificine of the genusTubificoides, a phallodriline of the genus Bathydrilus,nor in a number of marine enchytraeid genera and species found in Bermuda. The crystal-like structures have two needle arms, each about 5–10 m long and about 0.5 m in diameter, meeting at an obtuse angle. At the junction of the arms, there is a small membrane-bound `knob', about 1 m in diameter, which may be continuous with the coelomic peritoneum. The numbers of `crystals' per individual worm are estimated at 100–400 per body segment, or well over 2 × 10³ in an adult worm. `Crystals' are found: throughout the length of the worms, in all individuals of species in which `crystals' occur, and over the range of environmental conditions where these species are found in Bermuda. Simple digestions with hypochlorite, weak and dilute acids, and staining with nuclear and cytoplasmic stains indicate that the composition of the knob is organic and the arms inorganic. The fluorescent tracer Calcein (Sigma) was not incorporated into any structures during a 24-h bath incubation of living worms, and the `crystals' do not show birefringence when viewed between crossed polarizing filters. These last two results do not support an hypothesis that these are calcium carbonate `crystals'. Geographically, the crystal-like structures are widespread, and have also been observed in a species of immature (unidentified) marine tubificid from Rottnest Island, Western Australia.     

Some ultrastructural observations on the microfilaria of Breinlia sergenti-the pharyngeal thread and Innenkorper

Some ultrastructural observations on the microfilaria of Breinlia sergenti—the pharyngeal thread and Innenkorper.International Journal for Parasitology4: 375–382. The fine structure of the pharyngeal thread and the Innenkorper (inner body) of the microfilaria of Breinlia sergenti is described. The pharyngeal thread is seen as an elongated cuticular structure with associated nervous elements while the Innenkorper is sac-like in appearance. A possible function related to the transfer and storage of nutrient material is assigned to these structures.     

Some ultrastructural observations on the nature of foliar abscission in Impatiens sultani

Summary Both scanning and transmission electron microscopes have been used to study the anatomy of the abscission zone of Impatiens sultani Hook. Evidence is presented to show that the fracture line follows the middle lamella in all the living cells of the abscission zone including those in the vascular traces. The separation of these cells is preceded by a breakdown of the middle lamellar region of the wall. The characteristics of this process vary in different cell types. Accompanying this breakdown is an enlargement of inner cortex cells mainly in a direction parallel to the axis of the petiole. It is suggested that this expansion of cells is necessary to produce the tensions which rupture the cuticle and xylem vessels prior to separation. The occurrence of transfer cells and tyloses in the abscission zone is also described and the physiological implications of their presence discussed.     

Some observations on the use of cultured corneal endothelial cells as a model for intact corneal endothelium

Major differences have been identified between corneal endothelial cells in situ and those grown in culture. Cells in intact porcine corneal endothelium were studied and compared with primary cultures of the same cells either in suspension or in monolayers which had been grown on plastic (Nunc, Permonax). Differences were identified in the organization of the cytoskeleton (filamentous actin) between the cells in situ and in monolayer culture. The ability to withstand exposure to cryoprotective concentrations of Me(2)SO also varied substantially depending on whether the cells were in situ or in culture. These results underline the need for caution in the use of cells in culture as a model for studying the nature of injury to cells during the freezing of whole tissues.     

Identity, abundance and ecophysiology of filamentous Chloroflexi species present in activated sludge treatment plants

Filamentous Chloroflexi species are often present in activated sludge wastewater treatment plants in relatively low numbers, although bulking incidences caused by Chloroflexi filaments have been observed. A new species-specific gene probe for FISH was designed and using phylum-, subdivision-, morphotype 1851- and species-specific gene probes, the abundance of Chloroflexi filaments were monitored in samples from 126 industrial wastewater treatment plants from five European countries. Chloroflexi filaments were present in 50% of the samples, although in low quantities. In most treatment plants the filaments could only be identified with phylum or subdivision probes, indicating the presence of great undescribed biodiversity. The ecophysiology of various Chloroflexi filaments was investigated by a suite of in situ methods. The experiments revealed that Chloroflexi constituted a specialized group of filamentous bacteria only active under aerobic conditions consuming primarily carbohydrates. Many exo-enzymes were excreted, e.g. chitinase, glucuronidase and galactosidase, suggesting growth on complex polysaccharides. The surface of Chloroflexi filaments appeared to be hydrophilic compared to other filaments present. These results are generally supported by physiological studies of two new isolates. Based on the results obtained in this study, the potential role of filamentous Chloroflexi species in activated sludge is discussed.     

Some cultural and physiological aspects of methane-utilizing bacteria

A number of different methane-utilizing bacteria are described and compared with isolates of other investigators. The strains can be divided into three groups based on pigmentation, cell morphology and internal membrane structures. The oxidation of hydrocarbons, alcohols, aldehydes, fatty acids, methyl ethers and sugar phosphates by these bacteria was studied. There was much similarity between strains within the same group. Differences between groups as regards oxidative properties could be detected, but these were mainly quantitative and could not be used as taxonomical criteria. In addition, the inhibition of methane oxidation by metabolites and enzyme inhibitors was investigated. Formaldehyde proved to be the most active of the organic compounds tested. Iodoacetic acid inhibited both methane and methanol oxidation at concentrations of 0.03m or above. Of the inorganic compounds, KCN completely suppressed methane oxidation at 5×10^?4 m and to more than 90% at 5×10^?5 m.     

Some aspects on the role of bacteria in deep water ecosystems

Summary Some remarks on the role of bacteria in deep lakes will be presented with an emphasis on their functioning in the carbon cycle in such ecosystems.The metabolic intensity of most lakes is regulated to a large extent at the primary producer level. Aerobic and anaerobic decomposition by heterotrophic bacteria of the unstable ultimate products of photosynthesis results in the production of carbon dioxide and methane. Bacterial growth occurs at the expense of energy released by the flow of electrons from donors to acceptors. Typical electron acceptors for bacterially mediated reactions are oxygen, nitrate, sulphate and carbon dioxide. When oxygen is used as electron acceptor, the highest amount of energy is released, while the lowest is released when carbon dioxide is used. These reactions are mediated biologically, and the chemical reaction sequence is paralleled by an ecological succession of microorganisms: aerobic heterotrophs, denitrifiers, fermenting bacteria, sulphate reducers, and methane producers. The presence of oxygen is inhibitory to the organisms mediating the last reactions (STUMM, 1966; McCARTY, 1972), and this explains the succession of micro-organisms concomittant with the decrease in redox potential.Both labile and refractory fractions of the pelagial dissolved organic matter can directly be utilized largely by heterotrophic bacteria. At the onset of the summer stratification of Lake Vechten a high concentration (numbers 10⁹ bact/L) of heterotrophic bacteria has been detected in the lower water layers (CAPPENBERG, 1972). The reason for this increase may be a release of nutrients from the mud. During stratification the water layers above the mud become anaerobic owing to the metabolic activity of the bacterial flora and the chemical oxygen demand of the mud. In summer time the highest numbers of heterotrophic bacteria are found in the metalimnion, where organic matter accumulates due to a lower relative rate of sedimentation caused by an increase in viscosity and density.Furthermore it was found that sulphate-reducing bacteria which are capable of reducing sulphates to sulphides using sulphate as terminal electron acceptor, were observed in the hypolimnion only at the time of maximal stratification. During stagnation, gradually decreasing sulphate concentrations are found in the hypolimnion, correlated with increaseing cell numbers of sulphate reducers. As no sulphate could be detected in the mud of Lake Vechten as well, we may conclude that the sulphate concentration limits the number of these bacteria, as can be predicted for similar aquatic environments.Usually the sulphate reducers are found in bottom deposits and are an important group of bacteria of the non-methanogenic populations in mud. Summarizing the biological methane production and its subsequent oxidation by methane-oxidizing bacteria, we may conclude that these processes can be important factors functioning in the carbon cycle in deep fresh-water ecosystems.     

Some observations on phycobilisomes of Pterocladiella capillacea (Gelidiaceae,Rhodophyta)

ABSTRACT

Phycobilisomes (PBSs) of the red alga Pterocladiella capillacea collected in the field, were characterized both in situ and in vitro by means of a transmission electron microscope (TEM) and an image analyzer. Ultrathin sections of thalli and negatively stained PBSs after isolation revealed hemi-ellipsoidal shapes. In situ PBS dimensions were 38.5 ± 0.2 nm (height) × 38.8 ± 0.2 nm (width) × 22.6 ± 0.2 nm (thickness) in good agreement with the in vitro measurements (mean diameters of 37.6 ± 0.2 nm). These dimensions, especially the width, are smaller than those so far reported for red algae. This could depend either on the ecophysiological conditions of the thalli when harvested and/or on a staggered, symmetrically rotated and compressed disposition of biliprotein rods with respect to the allophycocyanin (APC) core. Hydroxylapatite chromatography of biliproteic extracts and SDS-PAGE electrophoresis revealed that phycoerythrin type R-(λ_max 565 nm>540 nm>498 nm) is formed by α (18.6 kDa), β (19.9 kDa), γ (30.2kDa) and γ′ (33.8 kDa) sub-units. The presence of two γ sub-units suggests that this phycoerythrin is a set of (αβ)₆γ + (αβ)₆γ′ aggregates (R-PE). A spectroscopically distinct form of phycoerythrin with different peak ratios, also found in pure fractions, is thought to be a polydisperse form (the so called r-PE). Similarity of shape and size observed in PBSs both in situ and in vitro, and fluorescence spectral characteristics of PBSs in vitro would indicate a substantial integrity of isolated PBSs. These measurements, if compared with total biliprotein content, would seem to indicate a PE fraction not assembled into PBS. A possible role of phycoerythrin in relation to the ability for a rapid adaptation of this surface species to environmental changes is suggested.     

Morphological and ultrastructural aspects of dehydration and rehydration in leaves of Sporobolus stapfianus

The resurrection species Sporobolus stapfianus Gandoger has been studied by LM, TEM and SEM in order to define the leaf morphology and fine structure and to analyse the cellular changes occurring during the processes of dehydration and rehydration of the plant. Some characteristics of the fully hydrated leaf and some ultrastructural and physiological events which take place during leaf wilting are discussed in relation to their possible role in plant desiccation-tolerance.The leaves of S. stapfianus show several characteristics common among xerophytic species. In the resurrection leaf they could play a role in slowing down the drying rate, thus leaving time to activate the mechanisms protecting the cell structures against drought damage. Actually, the S. stapfianus leaves do not undergo important cellular alterations during dehydration. The chloroplasts, in particular, retain part of their photosynthetic pigments and thylakoid membranes. Upon rewatering leaf recovery is rather fast and the tissue structure and cell organization of the fully hydrated state are already regained after two days.     

Characterization of digestive carbohydrase activity in the gilthead seabream (Sparus aurata)

Six species of Phallodrilinae are reported from Hainan Island in southern China. The mesopsammic Aktedrilus yiboi sp. nov. is new to science. It is characterized by a combination of (1) small, straight, conical, strongly cuticularized penis sheaths, (2) posterior prostate glands being associated with the most ectal parts of the atria, and (3) small spermathecal ampulla. Bathydrilus ampliductus Erséus, 1997 is recorded for the first time since it was originally described from Australia's Northern Territory.