Cover

ON THE COVER: Valve formation in a raphid diatom Diploneis smithii observed under SEM. The initial 2D network of ribs (top) thickens and begins vertical development (middle) to form an elaborate 3D structure with alternately arranged areolae, elliptical margin, and longitudinal canals on either side of the midrib containing the raphe slits. All valves are shown from the inside. [Vol. 54, No. 2, pp. 171–186 ]     

Ultrastructure of excretory system in <Emphasis Type="Italic">Bothrioplana semperi</Emphasis> (Platyhelminthes,Turbellaria)

The ultrastructure of flame bulbs and epithelium of excretory canals in Bothrioplana semperi (Turbellaria, Seriata) have been studied. The flame bulbs consist of two cells, the terminal cell and the proximal canal cell. The weir is formed by two rows of longitudinal ribs. The ribs of the internal row originate from the flame cell, external ribs are formed by the proximal canal cell. Each external rib has a remarkable bundle of microfilaments, originating in the cytoplasm of the first canal cell distally to the bases of external ribs. Membrane of internal ribs is marked by small electrondense granules, separate or fused to an electron-dense layer, continuous to dense “membrane,” connecting both external and internal ribs. Sparse internal leptotrichs originate from the bottom of the flame bulb cavity. External leptotrichs are lacking. Septate junction is present only in proximal canal cell at the level of tips of cilia. The apical surface of the canal cell bears rare short microvilli. The basal membrane of canal cells forms long invaginations that may reach nearly the apical membrane. The epithelium of excretory canals lacks the cilia. The ultrastructure of flame bulbs and epithelium of the excretory canals in B. semperi shares representatives of suborder Proseriata (Seriata). The contradiction exists in interpretation of the structure of flame bulbs in Proseriata. Ehlers and Sopott-Ehlers assumed that the external ribs are derivatives of the proximal canal cell and internal ones are outgrowths of the terminal cell, while Rohde has found conversely: the external ribs are outgrowths of the terminal cell, the internal ones are outgrowths of the proximal canal cell. However, the illustrations provided by Rohde do not enable to ascertain what cells the internal and external ribs derive from, while illustrations provided by Ehlers justify his interpretation. The order of weir formation in B. semperi confirms the viewpoint of Ehlers. The implication of ultrastructure of flame bulbs in Proseriata, especially of the order of flame bulb formation, in the Platyhelminthes phylogeny has been discussed.     

SCALY INCUNABULA,AUXOSPORE DEVELOPMENT,AND GIRDLE POLYMORPHISM IN SELLAPHORA MARVANII SP. NOV. (BACILLARIOPHYCEAE)1

Uniparental auxosporulation was observed in a monoclonal culture of a Sellaphora clone isolated from the epipelon of a fishpond in the Czech Republic. The cox1 sequence for the clone confirmed that it belonged to the Sellaphora pupula–bacillum species complex but showed significant differences from all previously characterized Sellaphora species, and it is therefore described as S. marvanii sp. nov. Protoplast, valve, and girdle structure resembled those of other Sellaphora species, but a novel finding for all diatoms was a change in girdle structure during the life cycle: the most advalvar girdle band (valvocopula) bore a single line of pores in enlarged postauxospore cells but was entirely plain in small cells and gametangia. The young auxospores were covered by incunabula containing large, delicate, ± circular scales, resembling those of centric diatom auxospores; similar scales have been reported in a few other raphid diatoms (Pseudo‐nitzschia multiseries, Diploneis sp.) but contrast with the strip incunabula of some Nitzschia and Pinnularia and the helmet‐like caps of Neidium. The scales persisted during auxospore expansion, mostly as two caps over the auxospore poles. The transverse perizonium comprised a very wide, closed primary band, flanked by numerous secondary bands whose open ends were strongly incurved toward the center. Initial valves were differentiated from their immediate descendants by the very strong external demarcation of the raphe sternum, irregular shape, and curved transapical profile.     

DIATOMS (BACILLARIOPHYTA) OF ISOLATED ISLANDS: NEW TAXA IN THE GENUS NAVICULA SENSU STRICTO FROM THE GALáPAGOS ISLANDS1

The diatoms (Bacillariophyta) from a coastal lagoon from the Diablas wetlands (Isla Isabela, the Galápagos Islands) were studied in material from surface samples and a sediment core spanning the past 2,700 years in order to examine evidence of diatom evolution under geographic isolation. The total number of taxa found was ～100. Ultrastructural variation in valve morphology between members of Galápagos taxa was used to describe 10 species from the genus Navicula sensu stricto, which are new to science. Four taxa: N. isabelensis, N. isabelensoides, N. isabelensiformis, and N. isabelensiminor, shared several key characteristics that may be indicative of a common evolutionary heritage; these species therefore provide possible evidence for the in situ evolution of diatoms in the Galápagos coastal lagoons. Shared morphological characteristics include: (i) stria patterning in the central area, (ii) an elevated and thickened external raphe‐sternum, (iii) external central raphe endings that are slightly deflected toward the valve primary side, and (iv) an arched valve surface. To explain these findings, two models were proposed. The first suggested limited lateral diatomaceous transport of Navicula species between the Galápagos and continental South America. Alternatively, these new species may be ecological specialists arising from the unique environmental conditions of the Galápagos coastal lagoons, which restrict the colonization of common diatom taxa and enable the establishment of novel, rare species. The Diablas wetlands are an important site for diatom research, where local‐scale environmental changes have combined with global‐scale biogeographic processes resulting in unique diatom assemblages.     

THE MORPHOLOGY AND INTEGRATION OF VALVES AND BANDS IN NAVICULA MUTICA VAR. MUTICA (BACILLARIOPHYCEAE)1

Navicula mutica (Kütz.) var. mutica was isolated from the air, cloned on agar, cultured in soil-water bottle, and studied with transmission and scanning electron micros-ropy. The frustules were lanceolate to ovoid with rounded apices, with the apical axis 8.5 ± 3.2 μ and the trans-apical and the transapical axis 3.6 ± 0.6 μm. Striae were composed of two or three puncta, and the mantle bore a single row of puncta aligned with the striae. The ends of the raphe turned away from an isolated punctual in the central area of the valve. The mantle puncta and one or two of the valve-face puncta in each stria opened into a series of transapical grooves in the interior of the valve, the grooves contributing to the appearance of striae in the light microscope. The interior of the mantle also possessed a pair of longitudinal grooves, discontinuous at the apices of the valves. An undulate advalvar margin of the valvocopula likely articulates along the interior longitudinal groove of the mantle. The projections of the undulate margin are perhaps positioned between the transapical grooves and along the longitudinal groove between the dentiform structures formed by the intersection of the double-grooved system. The girdle bands each had two (occasionally three) rows of pores. The pleurae margins were straight and not undulate.     

Ontogenetic analysis of siliceous cell wall formation in Triparma laevis f. inornata (Parmales,Stramenopiles)

Triparma laevis f. inornata is a unicellular alga belonging to the Bolidophyceae, which is most closely related to diatoms. Like diatoms, T. laevis f. inornata has a siliceous cell wall. The cell wall of T. laevis f. inornata consists of four round plates (three shields and one ventral plate) and one dorsal and three girdle plates. But, unlike diatoms, T. laevis f. inornata cells can grow when concentrations of silica are depleted. We took advantage of this ability, using TEM to study the ontogeny of the siliceous plate, pattern center formation, and development. Two types of pattern centers (annulus and sternum) were observed in the early and middle stage of plate formation. During their formation, the annuli were initially crescent‐shaped but eventually their ends fused to make a ring. Only outward silica deposition of the branching ribs occurred on the growing annulus until it became a ring, resulting in an unfilled circle inside the annulus. The pattern center of the shield plate was always an annulus, but in ventral plates both annulus and sternum were observed. The annuli and sterna in T. laevis f. inornata round plates were very similar to the annuli and sterna in diatom valves. These results suggested that the round plates of Parmales are homologous to diatom valves. This information on the plate ontogeny of T. laevis f. inornata provides new insights into the evolution of the siliceous cell wall in the Parmales and diatoms.     

TAXONOMY,LIFE CYCLE,AND AUXOSPORULATION OF NITZSCHIA FONTICOLA (BACILLARIOPHYTA)1

Nitzschia fonticola (Grunow) Grunow is a member of Nitzschia sect. Lanceolatae, a group of taxonomically intractable but ecologically important and widespread diatoms. We investigated the morphology and life cycle in three clones of N. fonticola and all exhibited reduced sexuality, with pedogamous production of auxospores in unpaired gametangia. The auxospores of all clones contained tangles of striplike elements that lay outside the perizonium and were distinct from it in structure and ontogeny. We introduce a new term, incunabula, to refer to such components of the auxospore wall. Semicryptic variation was detected: one clone differed from the other two in valve size and shape, stria density, and fibula density, as well as its nuclear large subunit ribosomal DNA (LSU rDNA) sequence. The implications of reduced sexuality for the taxonomy of sect. Lanceolatae are discussed. A lectotype is designated for N. fonticola from among original material of Grunow, and the application of the name is clarified further by designating illustrations and the LSU sequence AM182191 from one of our clones as epitypes.     

The composition and structure of the benthic diatom taxocene (Bacillariophyta) near Cape Fiolent (the Crimea,the Black Sea)

For the first time in the waters of the Cape Fiolent Reserve, a high species richness of benthic diatoms was revealed on natural substrates, 290 species and infraspecific taxa (IT), belonging to 281 species, 68 genera, 32 families, 19 orders, and 3 classes. A total of 68 species and 3 genera were previously registered as new for the flora of the Black Sea; 3 species and 1 taxonomic combination were previously described as new to science, as well as 4 species that did not occur in the Black sea in the past 50 and 100 years of research. Navicula (41 species and IT), Amphora (30), Nitzschia (29), Fallacia (20), Cocconeis (17) and Diploneis (16) were the most represented genera. Only one representative was identified in each of the orders Anaulales, Ardissoneales, Thalassionematales, Rhaphoneidales, and Rhopalodiales. The diversity assessment that was carried out using taxonomic distinctness indices, TaxDI, revealed that the structure of the phylogenetic diatom tree was formed by branches with various species saturation levels and different hierarchical subordination, but with the predominance of polyspecific taxa up to at genera level. The TaxDI values (Δ⁺ = 76.71; Λ⁺ = 361.52) indicate the low evenness and high variability of the structure of the benthic diatom taxocene near Cape Fiolent.     

Xenococconeis opunohusiensis gen. et sp. nov. and Xenococconeis neocaledonica comb. nov. (Bacillariophyta) from the tropical South Pacific

During a survey of the coral reef diatoms of Moorea Island (Society Archipelago, South Pacific) a small‐sized member of the order Achnanthales was studied using a light microscope (LM) and a scanning electron microscope (SEM). This marine taxon has: a raphe valve (RV) with a non‐crenulate edge; a high cingulum; a sternum valve (SV) often irregularly striated and areolae with concave hymenate pore occlusion; a thick and plain SV valvocopula (SVVC), ring‐shaped, composed of large fused fimbriae, with a central elliptic foramen bordered by the peg‐like edge of the fimbriae. On abvalvar side, the SVVC bears radiate concave and robust transapical ribs, interlinking with short elevated transverse ribs of the RV valvocopula (RVVC). Large marginal fenestrae of the RVVC give access to pseudoloculi. One oblong, unique and striated papilla is located on each RVVC rib. Given this unique set of features, we describe Xenococconeis opunohusiensis gen. et sp. nov. as a new taxon belonging to the Achnanthales. The characteristics of the new taxon are compared with Campyloneis Grunow and Cocconeis Ehrenberg. From New Caledonia, Cocconeis neocaledonica Maillard ex Lange‐Bertalot et Steindorf, a freshwater diatom, was described with two internal septa with marginal pseudoloculi. Based on subsequent SEM illustrations and remarks, we propose the transfer of C. neocaledonica to the new genus, and compare it to the type species, Xenococconeis opunohusiensis.     

Kulikovskiyia gen. nov. (Bacillariophyceae) from the lateritic rock pools of the Western Ghats,India and from Hainan Province,China

A new triundulate naviculoid diatom genus is described from the Western Ghats of Peninsular India and Hainan Province, China. The new taxon, Kulikovskiyia gen. nov. has robust conical spines along its margin and at the apices and the external valve face has longitudinally‐oriented siliceous slat system extending the length of the valve. The external distal raphe ends bifurcate and terminates on the valve face. There appear to be superficial similarities between this Asian genus and species and Playaensis, a genus comprised of two species found only in the western USA. The systematic position of Kulikovskiyia is discussed, and other than noting its similarities to other biraphid naviculoid diatoms due to symmetry features and the position of the raphe, we are uncertain about its systematic placement at finer levels of classification.     

OVERLOOKED HETEROPOLARITY IN SURIRELLA CF. FASTUOSA (BACILLARIOPHYTA) AND RELATIONSHIPS BETWEEN VALVE MORPHOGENESIS AND AUXOSPORE DEVELOPMENT

Surirella cf. fastuosa is an apparently isopolar elliptic marine raphid diatom. We observed cells before and after sexual reproduction in monoclonal cultures using light and scanning electron microscopy (LM and SEM). After sexual reproduction cells were approximately twice as large as before, in valve length and width. The stria and infundibula densities were stable during the life cycle. Subtle morphological differences were detectable between the two poles of the frustule. One pole (pole A) was characterized by endings of the external raphe fissure that turned toward the valve face, continuity of the domed wall of the raphe canal externally, an elliptic chamber visible internally, a shallow nick in the interior of the valvocopula. The other pole (pole B) was with the following: straight endings of the external raphe fissures, a dent in the domed wall of the raphe canal externally, a double chamber internally, presence of the open ends of the valvocopula nearby, a deep nick in the valvocopula. Furthermore, at pole A virgae developed at an early stage in morphogenesis, whereas at pole B they were not formed. In the auxospores, pole A was situated beneath the primary transverse perizonial band. Pole A is suggested to be homologous with the head pole in heteropolar Surirella and is the “protopole” likely equivalent to the central nodule in naviculoid taxa. Pole B is homologous with the foot pole in heteropolar Surirella and is the “synaptic pole” formed by fusion of components equivalent to both poles of naviculoid taxa.     

VALVE MORPHOGENESIS IN THE PENNATE DIATOM ACHNANTHES COARCTATA

Mitosis and valve morphogenesis in the pennate diatom Achnanthes coarctata (Bréb. in W. Sm.) Grun. are described. After cytokinesis, both daughter nuclei and their microtubule centers (MCs) are found near one side of the cell. Each new tubular silica deposition vesicle (SDV) arises centrally, forming a single rib running the length of the cell. Each MC then migrates around its nucleus and positions itself directly adjacent to the new SDV. The enlarging silicalemmas with their associated MCs, nuclei, microtubules (MTs) and microfilaments (MFs) appear in mirror image in the daughter cells. Both SDVs soon generate a second longitudinal rib alongside the first; the gap between the ribs ultimately becomes the future raphe fissure. The MC, MTs and nucleus are associated with each fissure. However, the subsequent behavior of the valve secreting machinery now becomes quite different in the daughter cells. In the cell that will form a raphid valve, the silicalemma, flanked by MFs, expands laterally in both directions over the cleavage furrow. Within the expanding SDV, silica secretion continues, eventually generating the structure of the mature valve, and during this phase the raphe fissure becomes delineated as in other raphid diatoms. In the other daughter cell, however, the MC and its MTs withdraw from the silicalemma, and the SDV moves laterally across the cleavage furrow until the double rib is at the corner of the cell. As silica is secreted into this expanding SDV, the raphe fissure completely fills in. This valve, therefore, lacks a raphe when mature and has a symmetry quite different from that of the valve formed in the other daughter cell. These events are compared with the course of morphogenesis described for other raphid diatoms.     

New species and new records of diatoms from Lake Fuxian,China

Abstract One new species, four newly recorded genera, and two newly recorded species of diatoms are described from China. They are Sellaphora fuxianensis sp. nov. Li, Sellaphora Mereschkowsky, Karayevia Round & Bukhtiyarova, Placoneis Mereschkowsky, Fallacia Stickle & Mann, Placoneis prespanensis Levkov, Krstic & Nakov, and Fallacia lucinensis (Hustedt) D. G. Mann. All taxa were collected from Lake Fuxian in Yunnan Province, China. These species were observed with light and scanning electron microscopes, and their taxonomic characters are discussed.     

The occipital region in the basal bony fish Erpetoichthys calabaricus (Actinopterygii: Cladistia)

Erpetoichthys calabaricus has unusual cranio‐vertebral anatomy, with an occipital centrum forming a component part of the compound basiexoccipital bone, and a ‘free‐floating’ occipital neural arch that differs from accessory arches found in some teleosts. The occipital neural arch bears autapomorphic lateral projections that articulate with small rod‐like bones resembling the spatial relationship of parapophyses and ribs, a feature normally restricted to vertebral centra. Based on analyses of cleared and stained specimens, computed tomography and histology, it is hypothesized that the lateral projections and associated rod‐shaped bones are structures that share developmental homologies to the unique ‘dorsal ribs’ of Polypteridae.     

THE “PARADOX” DIATOM BACILLARIA PAXILLIFER (BACILLARIOPHYTA) REVISITED1

Defined by its unique colonial locomotion, Bacillaria paxillifer (O. F. Müll.) Hendey was recognized as a single, pandemic species by many phycologists. However, reinvestigation of colonies from different habitats revealed three distinct groups: (A) brackish/freshwater, (B) marine littoral, and (C) marine planktonic taxa. Groups differed in colony and cell form, raphe flanges (RFs), shape and position of transapical ribs (Tr's), and morphogenesis. Linear‐shaped species were restricted to group A: Tr's thickened principally to the interior. Lanceolate forms were confined to groups B and C: valve formation proceeded from an internal base layer to the exterior. The planktonic species differed in the shape of its raphe slit, and the transformation of girdle bands (GBs) into “winglets.” Taxa also differed in chloroplast shape and number. All species formed motile colonies. Siblings adhered via elastic fibrils secreted through their raphe. Raphe ribs were held in position by siliceous clamps (fibulae), anchored in an extra pair of axial ribs (fibular ribs) parallel to the raphe ribs. This raphe system resembled that of Cylindrotheca rather than the “canal raphe” of Nitzschia. Many valves were asymmetric along the apical axis due to protruding RFs shuttling in a 1:1 ratio within a colony, but raphe slits were mirror images, as were the growth direction of fibulae and position of plastids, with pyrenoids tilted in the same direction. Species possessed four open GBs per epitheca; the third band invariably bore an internal, organic ridge to aid in adhesion of the plasmalemma during cleavage. The results suggested that these taxa are a natural phylogenetic group, requiring precise determination of their taxonomic position.     

Studies on the biochemistry and fine structure of silicia shell formation in diatoms VII. Sequential cell wall development in the pennateNavicula pelliculosa

Summary The development of the wall of synchronized culture ofN. pelliculosa is described. The first step, modification of the 3-2 configuration of the girdle bands of the wall during interphase, occurs immediately before mitotic division by the addition of a third girdl band to the hypotheca. Following cytokenesis, the new valve is initiated when a primary central band is formed within a silica deposition vesicle. This band extends the length of the cell and contains a central nodule. Secondary arms extend from the central nodule, join with extensions of the primary central band, and constitute the raphe rib. Mounds or knolls are formed on the central nodule and disappear as the valve matures. Transapical ribs appear on both the primary central band and secondary arms, and cross extensions join to form the sieve plate areas. The wall appears to be released by a joining of the inner silicalemma and the plasmalemma. An organic coat covers the newly released wall. Two girdle bands are formed and released sequentially.     

Intestinal Methanobrevibacter smithii but not total bacteria is related to diet‐induced weight gain in rats

It is increasingly understood that gastrointestinal (GI) methanogens, including Methanobrevibacter smithii, influence host metabolism.

Objective:

Therefore, we compared M. smithii colonization and weight gain in a rat model under different dietary conditions.

Design and Methods:

Sprague‐Dawley rats were inoculated with M. smithii or vehicle (N = 10/group), fed normal chow until day 112 postinoculation, high‐fat chow until day 182, then normal chow until day 253. Thereafter, five rats from each group were fed high‐fat and normal chow until euthanasia.

Results:

Both groups exhibited M. smithii colonization, which increased following inoculation only for the first 9 days. Change to high‐fat chow correlated with significant increases in weight (P < 0.00001) and stool M. smithii (P < 0.01) in all rats, with stool M. smithi decreasing on return to normal chow. Rats switched back to high‐fat on day 253 further increased weight (P < 0.001) and stool M. smithii (P = 0.039). Euthanasia revealed all animals had higher M. smithii, but not total bacteria, in the small intestine than in the colon. Rats switched back to high‐fat chow had higher M. smithii levels in the duodenum, ileum, and cecum than those fed normal chow; total bacteria did not differ in any bowel segment. Rats which gained more weight had more bowel segments colonized, and the lowest weight recorded was in a rat on high‐fat chow which had minimal M. smithii colonization.

Conclusions:

We conclude that M. smithii colonization occurs in the small bowel as well as in the colon, and that the level and extent of M. smithii colonization is predictive of degree of weight gain in this animal model.     

Morphology and ontogeny of multiple lateral‐line canals in the rock prickleback,Xiphister mucosus (Cottiformes: Zoarcoidei: Stichaeidae)

The structure and ontogeny of lateral‐line canals in the Rock Prickleback, Xiphister mucosus, were studied using cleared‐and‐stained specimens, and the distribution and morphology of neuromasts within lateral‐line canals were examined using histology. X. mucosus has seven cephalic canals in a pattern that, aside from four branches of the infraorbital canals, is similar to that of most teleostean fishes. Unlike most other teleosts, however, X. mucosus features multiple trunk lateral‐line canals. These include a short median posterior extension of the supratemporal canal and three paired, branching canals located on the dorsolateral, mediolateral, and ventrolateral surfaces. The ventrolateral canal (VLC) includes a loop across the ventral surface of the abdomen. All trunk canals, as well as the branches of the infraorbitals, are supported by small, dermal, ring‐like ossifications that develop independently from scales. Trunk canals develop asynchronously with the mediodorsal and dorsolateral canals (DLC) developing earliest, followed by the VLC, and, finally, by the mediolateral canal (MLC). Only the mediodorsal and DLC connect to the cephalic sensory canals. Fractal analysis shows that the complexity of the trunk lateral‐line canals stabilizes when all trunk canals develop and begin to branch. Histological sections show that neuromasts are present in all cephalic canals and in the DLC and MLC of the trunk. However, no neuromasts were identified in the VLC or its abdominal loop. The VLC cannot, therefore, directly function as a part of the mechanosensory system in X. mucosus. The evolution and functional role of multiple lateral‐line canals are discussed. J. Morphol. 276:1218–1229, 2015. © 2015 Wiley Periodicals, Inc.     

Valve morphogenesis in an araphid diatom Rhaphoneis amphiceros (Rhaphoneidaceae,Bacillariophyta)

The pattern centre in valve morphogenesis is an annulus in centric diatoms or a sternum in pennate diatoms. The genus Rhaphoneis is currently placed within a lineage that diverges at the root of the pennate diatom clade in most molecular phylogenies, and its valves have a unique pattern to their striae, i.e. radiating from both apices, giving the impression that a pattern centre exists at both ends of the valve and virgae (ribs) formation proceeds centripetally. The present study, however, shows that the pattern centre is actually a linear sternum and the formation of virgae proceeds centrifugally, a pattern centre that is commonly found in most araphid diatoms. Thus, the hypothesis that valve morphogenesis based on a linear sternum and perpendicular virgae is a synapomorphy of pennate diatoms is supported. Our study also demonstrates that the pattern of valve formation can be observed by light microscopy with a direct mounting method when the specimen is relatively large, i.e. exceeding approximately 50 µm in valve length. An important advantage of the use of the direct mounting method is that it requires no repeated centrifugation steps for dehydration, steps necessary for observation by a scanning electron microscope, causing the loss and/or collapse of the specimen, particularly with fragile valves in the early stages of development.