The primary body-wall musculature in the arrow-worm Sagitta setosa (Chaetognatha): An ultrastructural study

The primary musculature of Sagitta is mainly made up of two kinds of alternating fibers, A and B. These fibers differ markedly in their localization in the muscular tissue, by the development of their SR and their mitochondria, and the shape of their myofibrils. Their contractile apparatus is similar and possesses myofibrils of regular thickness with very short I bands, flanked by invaginations which are large compartments communicating with the extracellular space. This fiber diversity appears and is maintained in the presence of an apparent common innervation. Nerve endinglike structures are scattered in the epidermis against the basement membrane and there are no nerves beneath this. The presence of at least two kinds of fibers in the primary musculature and the presence of the secondary musculature would suggest that the displacements of sagitta may be more complex than is generally admitted. The specializations of the trunk musculature underline the degree of specialization in the Chaetognatha phylum.     

Ultrastructural study and ontogenesis of the appendages and related musculature of Paraspadella (Chaetognatha)

A lineage of benthic chaetognaths has developed limb-like appendages on the caudal part of the body, resulting from a local modification of the lateral fins, which are folds of the epidermis and have a role in balance when swimming. The most complex are those of Paraspadella gotoi which are used as props with the tip of the tail, allowing an elaborated mating behaviour comprising different movements: complete erection of the body, swings and jumps, astonishing for so simple-bodied animals. In the tail, the epidermis and the connective tissue, together with the longitudinal musculature, are involved in this innovation. All the components of the fins, i.e. connective tissue, fin rays and multilayered epidermic cells are conserved, but their function has changed. The movements of appendages are adjusted by one pair of small appendicular muscles localised in the body wall, while posture movements of the body are allowed by four longitudinal bundles of raising muscle. These two new muscles have successively appeared in the evolutive series previously described in Paraspadella. They have definitely arisen from the secondary muscle: the two lateral bundles for the former, and the two dorsal and two ventral ones for the latter. All are supercontracting muscles, a muscle kind also observed in the other benthic genus Spadella, but unknown in planktonic and benthoplanktonic chaetognaths.     

Ultrastructural and physiological studies of the contraction of the trunk musculature of Sagitta setosa (chaetognath)

The relationships between structure and function of the contractile apparatus of the trunk musculature in the chaetognath Sagitta setosa, were studied by electron microscopy and mechanical recordings. We also investigated the nature of the neuromediator at the synaptic level. Contraction, relaxation and stretch can be explained on the sliding model basis. The primary filaments are linked to a large Z line, via C filaments. These C filaments vary in length according to the mechanical states of the contractile apparatus. These muscles are directly excitable by electrical current, high K(+) solutions induced depolarization, or exogenous acetylcholine. The characteristics of the unitary contractile response (twitch) and the effects of the frequency of stimulation set up the contractions of Sagitta among the fastest in the animal kingdom. Acetylcholine appears to be the best candidate for the mediator released at the neuromuscular junction. Its effect on the post-junctional membrane seems to require a receptor of the nicotinic type.     

Laboratory feeding and egg production in the chaetognath Sagitta crassa Tokioka

Egg production, daily ration, and growth efficiency in Sagitta crassa Tokioka are described along with the morphological characteristics of the ovary and body during laboratory observations. Feeding by S. crassa in small containers containing 50 or 100 ml sea water and feeding rates bore a linear relationship to food concentration from 5 to 20 copepods per 100 ml. Among 100 animals individually isolated in containers, 3 animals were the longest lived and the best egg producers. One of these produced eggs on more than 30 consecutive days, producing almost 1000 eggs. Egg production showed periodic change at intervals from 7 to 10 days. Seminal vesicles also became periodically filled or empty. Fluctuations in ovary length and morphological changes of ovary and body were observed through life. Daily ration of S. crassa ranged from 8.7 to 10.4 prey per day or ≈ 60 μg in dry wt. The specific daily ration ranged from 0.347 to 0.568. S. crassa had a growth efficiency (dry-wt basis) which increased during the early life of the animal (≈28%), then decreased with age (22 → nearly 0%). Reproductive efficiency ranged from 7.0 to 16.4%, with a mean of 12.5%.     

Ultrastructural studies on the microfilaria of Cardianema sp. Alicata, 1933 (Nematoda; Onchocercidae)

The ultrastructure of the sheath, cuticle and hypodermis of the microfilaria of Cardianema sp, is described from electron micrographs of in utero- and blood-stages. The trilaminiar sheath invests the microfilaria throughout development in utero and it acquires a superficial coat after the microfilaria enters the blood stream of its reptile host. The cuticle consists of external and internal cortex, fibrillar and subfibrillar layers. The cuticle is attached to the hypodermis without the intervention of a basal lamina. The structure of the external cortex is modified in the annular furrows in the cuticle. The cellular hypodermis forms a complete subcuticular layer, although over much of the circumference the cells exist as thin cytoplasmic processes and where these overlap there are extensive tight junctions. The case for classifying the microfilaria of Cardianema a first stage larva is advanced and a functional but speculative, role for the sheath is proposed.     

Ultrastructural observations on Barroussia schneideri (apicomplexa,eucoccidiida) in the centipede Lithobius forficatus

The ultrastructure of the principal stages of Barroussia schneideri in the intestinal cells of the centipede, Lithobius forficatus, is described. The structure of the merozoites has much in common with that of other Eimeriidae: pellicle of an outer and two inner membranes, microtubules, mitochondria, micronemes, rhoptries, endoplasmic reticulum, and polysaccharide granules. The mature macrogamete has a large nucleus with a distinct nucleolus, wall-forming bodies of type 1 (WFB1), and presumably of type 2 (WFB2), and a large number of polysaccharide granules. The microgametocyte includes smaller polysaccharide granules closely associated with the cisternae of the endoplasmic reticulum. The microgametocyte surface is smooth in outline and has peripherally arranged nuclei. Microgametes have a curved nucleus, a mitochondrion, polysaccharide granules, and two flagella. One of the flagella is attached for some distance along the body.     

The anterior glands of adult Necator americanus (Nematoda: Strongyloidea)—I Ultrastructural studies

The ultrastructure of the amphidial, oesophageal and excretory glands of N. americanus is described. There are two amphidial glands, and each is attached to a lateral hypodermal cord. Anteriorly the glands become associated with the amphidial sense organs. The amphidial glands synthesize complex secretion granules which appear to release their contents into the sense organ. Secretions thus pass over the amphidial cilia and exit via the amphidial pore. It is suggested that the secretory activity of these glands is under direct nervous control. There are three oesophageal glands, and each synthesizes dense secretion granules. The secretions of the oesophageal glands are released into the lumen of the oesophagus and into the buccal capsule. The two excretory glands are ventral in position and connected to the tubular excretory system. These glands synthesize secretion granules of varying density. Secretions from the excretory glands may exit via the excretory pore, or pass back into the tubular excretory system, or both.     

Ultrastructural observation of oogenesis in the crustacea amphipoda Orchestia gammarellus (Pallas)

The oogenesis of the Crustacea Amphipoda Orchestia gammarellus can be divided in five stages taking into consideration both the oocyte ultrastructure and the physiology of the ovary. The primary oogonium (12 μm in diameter) is lodged within the germinative zone: after division, the daughter cell (or secondary oogonium) leaves this area and enters meiotic prophase. Stage I is represented by the oocyte with visible chromosomes (12–18 μm in diameter) the cytoplasmic ultrastructure of which is comparable to that of the oogonium. Stage II or previtellogenesis is characterized by a considerable growth of the oocyte (18–80 μm in diameter) which becomes enriched in ribosomes and vesicles of the rough endoplasmic reticulum; the oocyte does not yet contain any vitelline reserve (proteinaceous and lipid). Stage III or primary vitello-genesis (80–160 μm in diameter) is typified by the synthetic activity of the rough endoplasmic reticulum, corresponding to an endogenous accumulation of proteinaceous yolk. Stage IV or secondary vitellogenesis (160–800 μm in diameter) only appears during the period of reproduction; by means of endocytosis the oocyte accumulates yolk spheres in addition to lipid droplets, the origin of which is uncertain; towards the end of vitellogenesis, cortical granules become a feature that is noted for the first time in Crustacea. The last stage or maturation (800 μm in diameter) starts right before or immediately after the exuviation of the female and ends with fertilization.     

Septal pore complex morphology in the Agaricomycotina (Basidiomycota) with emphasis on the Cantharellales and Hymenochaetales

The ultrastructure of septa and septum-associated septal pore caps are important taxonomic markers in the Agaricomycotina (Basidiomycota, Fungi). The septal pore caps covering the typical basidiomycetous dolipore septum are divided into three main phenotypically recognized morphotypes: vesicular-tubular (including the vesicular, sacculate, tubular, ampulliform, and globular morphotypes), imperforate, and perforate. Until recently, the septal pore cap-type reflected the higher-order relationships within the Agaricomycotina. However, the new classification of Fungi resulted in many changes including revision of existing and addition of new orders. Therefore, the septal pore cap ultrastructure of more than 325 species as reported in literature was related to this new classification. In addition, the septal pore cap ultrastructures of Rickenella fibula and Cantharellus formosus were examined by transmission electron microscopy. Both fungi have dolipore septa associated with perforate septal pore caps. These results combined with data from the literature show that the septal pore cap-type within orders of the Agaricomycotina is generally monomorphic, except for the Cantharellales and Hymenochaetales.It appears from the fungal phylogeny combined with the septal pore cap ultrastructure that the vesicular-tubular and the imperforate type both may have arisen from endoplasmic reticulum. Thereafter, the imperforate type eventually gave rise to the perforate septal pore cap-type.     

Further studies on the junctional complex in the intestine of Sagitta setosa

Summary The intramembrane structures of the pleated septate junction which occur in the junctional complex of the intestine of the chaetognath Sagitta setosa have been investigated.The pleated septate junction is made up of linear rows of irregularly shaped and sized particles, often fused into short rods, and pits which can be fused into furrows. The distribution of these structures on E and P faces depends upon the preparative methods used. Many of the morphological characteristics are the same as those of the lower invertebrate pleated septate junction type defined by Green (1981a). The physiological significance of this junction is obscure.On the basis of the presence of septate junctions (both of the paired septate junction and pleated septate junction types) which have mainly morphological characteristics of the lower invertebrate pleated septate junction we can add to the hypothesis that chaetognaths are not related to the molluscs and arthropods.     

Immunohistochemical and ultrastructural studies on ciliary sense organs of arrow worms (Chaetognatha)

We examined epibenthic and pelagic species of Chaetognatha (Spadellidae and Sagittidae) using immunohistofluorescence and confocal laser scanning microscopy to detect tubulin and cell nuclei in whole-mount preparations and scanning and transmission electron microscopy to visualize the ultrastructural organisation of their ciliary sense organs. All chaetognaths bear three types of ciliary sense organs distributed throughout the body: (1) transversally oriented ciliary fence organs, (2) longitudinally (parallel to the anterior-posterior axis) oriented ciliary tuft organs, and (3) a ciliary loop, the corona ciliata. This study targets the ciliary fence as well as the ciliary tuft organs. Two types of primary receptor cells constitute the ciliary fence and ciliary tuft organs. The first type of receptor cells forms a single cell line along the midline axis of the organs, whereas the second type of receptor cells forms multiple lines of cells at either side of type 1 cells. Each receptor cell extends a single, non-locomotory cilium from its narrow apex collared by slender, non-reinforced microvilli; however, both types of sensory cells considerably differ on ultrastructural level. Type 1 sensory cells have thicker cilia than those protruded by the type 2 sensory cells which are characterized by rootlets consisting of an elongated, amorphous distal as well as a cross-striated proximal portion. These results likely reveal that both types of sensory cells have distinct functions.     

Three-dimensional reconstruction of the musculature of Cossura pygodactylata Jones, 1956 (Annelida: Cossuridae)

The musculature of adult specimens of Cossura pygodactylata was studied by means of F-actin labelling and confocal laser scanning microscopy (CLSM). Their body wall is comprised of five longitudinal muscle bands: two dorsal, two ventral and one ventromedial. Complete circular fibres are found only in the abdominal region, and they are developed only on the border of the segments. Thoracic and posterior body regions contain only transverse fibres ending near the ventral longitudinal bands. Almost-complete rings of transverse muscles, with gaps on the dorsal and ventral sides, surround the terminal part of the pygidium. Four longitudinal bands go to the middle of the prostomium and 5–14 paired dorso-ventral muscle fibres arise in its distal part. Each buccal tentacle contains one thick and two thin longitudinal muscle filaments; thick muscle fibres from all tentacles merge, forming left and right tentacle protractors rooted in the dorsal longitudinal bands of the body wall. The circumbuccal complex includes well-developed upper and lower lips. These lips contain an outer layer of transverse fibres, and the lower lip also contains inner oblique muscles going to the dorsal longitudinal bands. The branchial filament contains two longitudinal muscle fibres that do not connect with the body musculature. The parapodial complex includes strong intersegmental and segmental oblique muscles in the thoracic region only; chaetal retractors, protractors and muscles of the body wall are present in all body regions. Muscle fibres are developed in the dorsal and ventral mesenteries. One semi-circular fibre is developed on the border of each segment and is most likely embedded in the dissepiment. The intestine has thin circular fibres along its full length. The dorsal blood vessel has strong muscle fibres that cover its anterior part, which is called the heart. It consists of short longitudinal elements forming regular rings and inner partitions. The musculature of C. pygodactylata includes some elements that are homologous with similar muscular components in other polychaetes (i.e., the body wall and most parapodial muscles) and several unique features, mostly at the anterior end.     

The pre-cephalic development of the skeleton of Amphimelissa setosa (Actinopoda: Nassellarida)

The ontogeny of the skeleton of the nassellarian radiolarian Amphimelissa setosa begins with the formation of three radial bars of silica; these correspond to the medial bar and spines L₁ and L₂ in the adult. By a precise sequence of the addition of various siliceous arcs and spines, this basic structure develops into the eucephalus. A new growth mechanism, in which existing skeletal elements serve as templates for the formation of new skeletal members, is hypothesized to explain the observed developmental patterns. The early ontogeny of nassellarian skeletons can be used to determine which skeletal features develop earliest. Among the structural characteristics used by radiolarian systematists, these conservative features are most suitable for use in interpretation of the taxonomic relationships and phylogeny of the group.     

The use of bis(mercaptoacetato-S,O)hydroxoiron(III) complex for the determination of hydroxamates

A rapid, indirect, spectrophometric procedure for the determination of hydroxamates, based on the competition for ferric ions of the bis(mercaptoacetato-S,O)hydroxoiron(III) complex, has been developed. The assay is remarkably free of interferences by common ions, thus rendering it useful in the quantitative determination of hydroxamates in culture fluids and crude preparations.     

Micropropagation and bioreactor studies of the medicinally important plant Lessertia (Sutherlandia) frutescens L.

This study describes a protocol for rapid and efficient in vitro propagation of Lessertia frutescens (cancer bush), a medicinally important plant species native to southern Africa. Single node explants were grown in various culture regimes of MS medium containing 30 g/l sucrose supplemented with various concentrations of cytokinins and auxins and solidified with 8 g/l agar. These were (a) 2.22, 4.44, 13.32 and 22.19 µM BA; 2.32, 4.65, 13.95 and 23.23 µM K and 0.45, 2.27, 4.54 and 13.62 µM TDZ (b) a combination of 2.22 µM BA with 0.57, 2.85, 5.71 and 11.42 µM IAA, 0.49, 2.46, 4.9 and 9.8 µM IBA or 0.54, 2.69, 5.37 and 10.74 µM NAA and (c) different media types viz. MS, SH basal salt medium and WPM at 1, ½ and ¼ salt strength which were each supplemented with 2.22 µM BA and 0.54 µM NAA. Single node explants were also grown in MS liquid medium supplemented with 2.22 µM BA and 0.54 µM NAA in temporary and continuous immersion bioreactors. Maximum number of shoots (12.9) per single node explant was obtained in the temporary immersion bioreactor but 50% of these shoots showed symptoms of hyperhydricity. In solid culture the best shoot multiplication response (10 shoots) was obtained in full strength MS. Roots were induced using shoot tips cultured in ½ MS solid medium supplemented with various concentrations of IBA or NAA. The highest rooting percentage (78%) was achieved in 19.6 µM IBA. Rooted plantlets were cultured in a mixture of perlite and vermiculite (1:1; v/v) and successfully acclimatized in a growth chamber with an 85% survival rate.     

Structural studies of sapote (Sapota achras) gum

Sapote gum contains residues of L-arabinose (pyranose and furanose), D-xylose, D-glucuronic acid, and 4-O-methyl-D-glucuronic acid in the ratio 1.0:2.8:0.48:0.52. The two uronic acids were conveniently determined by reducing the carboxyl functions with lithium borohydride and measuring the ratio of D-glucose to 4-O-methyl-D-glucose. Periodate oxidation of the carboxyl-reduced gum gave inter alia 2-O-methyl-D-erythritol and 4-O-methyl-D-glucose in amounts suggesting that 37% of the 4-O-methyl-D-glucuronic acid residues are unsubstituted in the polysaccharide. Acetolysis of the carboxyl-reduced gum gave O-α-D-glucopyranosyl-(1→2)-(4-O-β-D-xylopyranosyl)_0,1,2,-D-xylose, a hitherto undescribed series of oligosaccharides, together with 2-O-(4-O-methyl-α-D-glucopyranosyl)-D-xylose. Methylation confirmed that sapote gum has a highly branched structure, and commercial xylanases did not depolymerize the gum. An α-L-arabinofuranosidase liberated a substantial part of the arabinose residues. Sapote gum is a member of the uncommon class of plant gums having a D-xylose backbone and structurally resembles brea gum.     

Ultrastructural changes in salivary glands of Pseudolynchia canariensis (Dipteria: Hippoboscidae) infected with sporozoites of Haemoproteus columbae

Salivary gland cells of Pseudolynchia canariensis are ultrastructurally similar to those of many other diptera. Heavy infections with Haemoproteus columbae sporozoites cause disruption of the basement membrane, basal plasma membrane, and disorganization of the functional organelles of the cell. Cytoplasmic microtubules appear to increase in number.     

Ultrastructural studies on the vitellogenesis of Tetrodontophora bielanensis (Waga) (Collembola)

Summary Vitellogenesis in Tetrodontophora bielanensis (Waga) is of the mixed type. Part of the yolk material is produced inside the oocyte (auto-synthesis), while part is absorbed by micropinocytosis. During autosynthesis polyribosomes, rough endoplasmic reticulum and dictyosomes take part. Regardless of their origin, mature yolk spheres are constructed identically and are composed of three elements: cortex layer, matrix and crystals. Histochemical tests show that polysaccharides are present in the yolk spheres. Lipid droplets have been observed in the ooplasm; they develop without visible contact with any of the organelles. Among the reserve materials the following have been found: rough endoplasmic reticulum, dictyosomes, polyribosomes, mitochondria and a few microtubules.