Phylogenetic position and morphology of Raphidiophrys elongata sp. nov. (Haptista: Centroplasthelida) with notes on cyst wall structure and evolution

The majority of centrohelids bear external coverings consisting of organic spicules or siliceous scales. Cyst coverings are usually reinforced with additional layers of modified scales. The cyst wall of Raphidiophrys heterophryoidea has an unusual and complex structure. It consists of three different types of scales and includes the mosaic scale layer not known in other centrohelids. During excystment, the cyst wall fragments along the sutures of the mosaic layer. For other Raphidiophrys species, cyst coverings are not studied. The present paper describes a new Raphidiophrys species, R. elongata, belonging to the NC7 environmental clade. Trophozoites bore thin plate scales with reduced upper plate. Under starvation, cysts emerged in clonal cultures. Cyst coverings of R. elongata and R. heterophryoidea were studied in comparison with the use of FIB-SEM. Cyst wall of R. elongata was significantly thinner than in R. heterophryoidea and was formed with 3–5 layers of uniform overlapping scales. No mosaic scale layer was present. During excystment, trophozoite exited cyst shell through random fissure. Possible evolutionary events and driving forces behind the complication of cyst wall within Raphidiophrys were discussed.     

Characterization of brine shrimp Artemia from Cam Ranh Bay in Central Vietnam

Artemia cysts collected from inoculation experiments in Cam Ranh salterns are evaluated for their potential use in aquaculture. Cyst biometrics, hatching quality, naupliar fatty acid profile and naupliar growth were measured and compared to reference Artemia strains. Cyst characteristics reveal that the parthenogenetic strain (PR China) used in inoculations, was eliminated from the environment and that the remaining brine shrimp are likely to be composed of Macau and Great Salt Lake Artemia strains, and of their cross-breds. Differences in cyst diapause deactivation characteristics between Macau and Great Salt Lake Artemia may have resulted in the disappearance of Macau Artemia during the rainy season and the persistence of Great Salt Lake Artemia during the following dry season.     

Artemia monica cyst production and recruitment in Mono Lake,California, USA

Annual egg production was determined for Artemia monica in Mono Lake, California, from 1983 to 1987. Annual oviparous (overwintering cyst) production was 3 and 7 million cysts m^–2 yr^–1 in 1986 and 1987, respectively, as measured by in situ sediment traps. Cyst production for the entire five year period was calculated using Artemia census data and inter-brood duration derived from mixolimnetic temperature. These estimates ranged from 2 to 5 million cysts m^–2 yr^–1. This method underestimated annual production by 30%, when compared to estimates using sediment traps. Cyst production was similar during 1983–1986 and showed a significant increase in 1987, which was due primarily to a larger reproductive population later in the year. Recruitment into the adult populations of the following spring ranged between 1.4 to 3.2%. Overall abundance of this generation reflected the patterns in annual cyst production. Compensatory effects must operate on the second generation of each year, since summer populations were similar in all years despite differences in cyst production.     

Resting Cysts of the Pigmented Ciliate Blepharisma sinuosum Sawaya, 1940 (Ciliophora: Heterotrichea)

The morphology of Blepharisma sinuosum resting cysts and the dynamics of pigmentation at different stages of encystment are presented for the first time. Cyst morphometrics are similar to other Blepharisma species, with three‐wall layers, bacteria surrounding the ectocyst, a conical plug, and wrinkly surface toward the plug in mature stages. The vegetative moniliform macronucleus changes to a horseshoe shape, and at early stages, the cystic cytoplasm is homogeneously pigmented, comprising a contractile vacuole; later, pigments polarize toward the plug, decorate the cortical layer, and become brownish. This work reinforces the potential role of pigment dynamics on cyst biology.     

The Fine Structure of Secretion in Hyalophysa chattoni: Formation of the Attachment Peduncle and the Chitinous Phoretic Cyst Wall

ABSTRACT. The settling tomite stage of the apostome Hyalophysa chattoni secretes a phoretic cyst wall composed of chitin, mucopolysaccharides, and protein. Within 1 1/2 h after settling, an electron-dense proteinaceous cyst layer (the outer layer) is formed from secretions originating at the base of the kineties and from the thick pellicular layer between the kineties. The inner cyst layer, composed primarily of chitin (acidic and neutral polysaccharides are also present), is secreted across the entire cell surface. Cyst wall formation is completed within 6 h. The fine structure of endocyst secretion resembles stages in the secretion of chitin by fungi, yeasts, and arthropods. A proteinaceous attachment peduncle is secreted to anchor the cell to a shrimp host and is formed by the release of electron-dense secretory bodies from the cell's ventral surface.     

A Key and Compendium to Species of the Heterodera avenae Group (Nematoda: Heteroderidae)

A key based on cyst and juvenile characters is given for identification of 12 valid Heterodera species in the H. avenae group. A compendium providing the most important diagnostic characters for use in identification of species is included as a supplement to the key. Cyst characters are most useful for separating species; these include shape, color, cyst wall pattern, fenestration, vulval slit length, and the posterior cone including presence or absence of bullae and underbridge. Also useful are those of second-stage juvenile characteristics including aspects of the stylet knobs, tail hyaline tail terminus, and lateral field. Photomicrographs of diagnostically important morphological features complement the compendium.     

Localization of chitin on ultrathin sections of cysts of two ciliated protozoa,Blepharisma undulans andPseudomicrothorax dubius,using colloidal gold conjugated wheat germ agglutinin

Summary The cyst walls of the ciliatesBlepharisma undulans andPseudomicrothorax dubius were examined ultrastructurally and by postembedding labeling with wheat germ agglutinin (WGA)-gold conjugate. Different methods of fixation and embedding were performed. In all procedures, WGA-gold binds selectively to material of the cyst wall. Pretreatment of the sections with chitinase inhibits labeling. The cyst walls of both species contain 3 nm fibrils, which are supposed to be of chitinous nature. In the cyst wall ofB. undulans, several thin layers of WGA-binding fibrils are interspaced with thick layers of other material. InP. dubius, WGA-binding sites are mainly concentrated in the mesocyst, where the microfibrils appear to represent the major component. These results obtained from two phylogenetically distant species confirm that chitin synthesis is an ancestral feature of ciliated protozoa. The amount and distribution of the chitin fibrils may play an important role in the properties and functions of the wall of the resting cyst.     

Formation of sperm cells inGalium mollugo (Rubiaceae),Trichodiadema setuliferum (Aizoaceae), andAvena sativa (Poaceae)

The formation of sperm cells has been examined ultrastructurally in the tricellular pollen grains ofGalium mollugo L. (Rubiaceae).Trichodiadema setuliferum Schwantes (Aizoaceae), andAvena sativa L. (Poaceae). After detachement from the intine the generative cell of all three species lies free within the vegetative cytoplasm. The two sperm cells are built inTrichodiadema andAvena by a single separating wall, while inGalium mollugo two independent walls are formed. However, both mechanisms separate the two male gametes completely.     

AN ENCYSTMENT STAGE,BEARING A NEW SCALE TYPE,OF THE ANTARCTIC PRASINOPHYTE PYRAMIMONAS GELIDICOLA AND ITS PALEOLIMNOLOGICAL AND TAXONOMIC SIGNIFICANCE1

Cysts of the Antarctic prasinophyte Pyramimonas gelidicola McFadden were found in water samples from a fjord and a saline lake in the Vestfold Hills, Antarctica Unialgal cultures of P. gelidicola from Ace Lake produced cysts. After ca. five weeks, tile cysts settled and adhered to the bottom of the culture flask. The cyst wall was covered by a scale type not seen on the flagellated cells; however, the base of the cyst scale was similar to the box scales of P. gelidicola motile cells. Cyst scales were also found off the continental shelf in Prydz Bay. In a 1.7 m sediment core taken from Ace Lake, both cyst scales and box scales of P. gelidicola occurred at most depths. Differences in the ratio of these two scale types at different depths in the core may indicate past ecological changes in the lake. Upper sediments of the core were dated at 5310 ± 90 yrs B.P., indicating that prasinophyte scales may be recognizably preserved for extended periods. P. gelidicola was widely distributed in saline lakes of the Vestfold Hills with salinities of 3.2–133% and temperatures ranging from – 5.0 to 10.4°C. This is the first report of encystment of P. gelidicola and, to our knowledge, is the first record of a prasinophyte with two distinctly different scale types occurring on cells during different stages of the life history.     

SEASONAL VARIABILITY OF THE ORGANIC‐WALLED DINOFLAGELLATE CYST PRODUCTION IN THE COASTAL UPWELLING REGION OFF CAPE BLANC (MAURITANIA): A FIVE‐YEAR SURVEY1

A 5‐year sediment trap survey in the upwelling area off Cape Blanc (NW Africa) provides information on the seasonal and annual resting cyst production of dinoflagellates, their sinking characteristics and preservation potential. Strong annual variation in cyst production characterizes the region. Cyst production of generally all investigated species, including Alexandrium pseudogonyaulax (Biecheler) T. Horig. ex T. Kita et Fukuyo (cyst genus Impagidinium) and Gonyaulax spinifera (Clap. et J. Lachm.) Diesing (cyst genus Nematosphaeropsis) was enhanced with increasing upper water nutrient and trace‐element concentrations. Cyst production of Lingulodinium polyedrum (F. Stein) J. D. Dodge was the highest at the transition between upwelling and upwelling‐relaxation. Cyst production of Protoperidinium americanum (Gran et Braarud) Balech, Protoperidinium monospinum (Paulsen) K. A. F. Zonn. et B. Dale, and Protoperidinium stellatum (D. Wall) Balech, and heterotrophic dinoflagellates forming Brigantedinium spp. and Echinidinium aculeatum Zonn., increased most pronouncedly during upwelling episodes. Production of Protoperidinium conicum (Gran) Balech and Protoperidinium pentagonum (Gran) Balech cysts and total diatom valves were related, providing evidence of a predator–prey relationship. The export cyst‐flux of E. aculeatum, P. americanum, P. monospinum, and P. stellatum was strongly linked to the flux of total diatom valves and CaCO₃, whereas the export production of Echinidinium granulatum Zonn. and Protoperidinium subinerme (Paulsen) A. R. Loebl. correlated with total organic carbon, suggesting potential consumption of diatoms, prymnesiophytes, and organic matter, respectively. Sinking velocities were at least 274 m · d^?1, which is in range of the diatom‐ and coccolith‐based phytoplankton aggregates and “slower” fecal pellets. Species‐selective degradation did not occur in the water column, but on the ocean floor.     

CYST FORMATION IN THE FRESHWATER DINOFLAGELLATE CERATIUM HIRUNDINELLA (DINOPHYCEAE)1

Cyst formation in Ceratium hirundinella (O. F. Müll.) Bergh was studied by light and electron microscopy, using material from several lakes and reservoirs and also laboratory cultures. Cells preparing to encyst build up large quantities of starch and lipid and at the same time reduce their other cell components. The cyst is released from the theca as a naked cell bounded by a double membrane. The most commonly found cyst deposits a layer of electron-dense granules containing silicon on the outer membrane and lays down a cellulose-like material between the two membranes. Cysts without the electron-dense granules are commonly formed in cultures but rarely found in lakes. These cysts appear less resistant to decay and do not show the reorganization of cell contents for dormancy. It is suggested that C. hirundinella has both a resting cyst, forming part of the life cycle, and a temporary cyst stage.     

Protective role of astaxanthin against u.v.-B irradiation in the green alga Haematococcus pluvialis

Cyst cells of the green alga Haematococcus pluvialis accumulate astaxanthin with maturation of the resting stage. To study the protective role of astaxanthin against u.v. damage, both immature (astaxanthin-poor) and mature (astaxanthin-rich) cyst cells were exposed to u.v.-A or u.v.-B irradiation, and the residual cell viability and astaxanthin levels were determined. u.v.-B decreased both cell viability and astaxanthin level of cyst cells to a greater extent than u.v.-A. Tolerance of mature cyst cells to u.v.-B was 6-fold higher than that of immature cyst cells. These results indicated that astaxanthin in cyst cells functions as a protective agent against u.v.-B irradiation.     

The biology ofChlamydomyxa montana: Ultrastructure of the cyst

Summary Cells ofChlamydomyxa montana Lankester photosynthesize within a cyst under bright light and ingest diatoms in an excysted amoeboid form during periods of low light intensity. Cyst cell walls are partially comprised of cellulose and vary in thickness. The cells are multinucleate and intracellular bacteria are closely associated with the nuclei. A pair of centrioles is also present adjacent to individual nuclei. Chloroplasts are numerous and thylakoids are generally organized into bands of three. No endoplasmic reticulum was found surrounding the plastids, nor was a complete girdling lamella ever observed within.C. montana chloroplasts appear to be its own but this protist does not precisely fit into an algal class.     

A light and electron microscopic study of tissue interactions between a parasitic copepod, Scolecodes huntsmani (Henderson), and its host ascidian, Styela gibbsii (Stimpson)

The structure of the cellular cyst which encapsulates the parasitic copepod, Scolecodes huntsmani, in the subendostylar blood vessel of the ascidian, Styela gibbsii, is described from light and electron microscopic studies. The cells comprising the cyst are contributed by the ascidian. The cells are columnar, contain large central reservoirs of glycogen and lipid, and have a conspicuous Golgi apparatus, many small cisternae of smooth endoplasmic reticulum and peripheral mitochondria. The cells are held together by complex basal interdigitations and a short apical zonula occludens. Long cilia emerge in circular clusters from the cell apices and beat in the lumen of the cyst. As atypical of a columnar epithelial layer, the nuclei are staggered in position in the cells and there is no basal lamina. One end of the cyst is blind, but the other end, which may be either anterior or posterior with respect to the longitudinal axis of the host, narrows to a profusely ciliated duct which opens through the wall of the blood vessel to the atrium of the ascidian by a ciliated funnel. The effective beat of the cilia of the duct and the funnel is outward toward the atrium. The first nauplii of the copepod emerge from the incubatory pouch of the adult and pass to the exterior sea water through the cyst funnel and the atrium and atrial siphon of the ascidian. As in other notodelphyid copepods, the life cycle of this incarcerated form also involves free-living naupliar stages followed by two free-living copepodid stages. The provision of an egress for the first nauplii is, therefore, important to the survival of the species. The adult females of Scolecodes, which range in length from 2 to 14.6 mm, are sluggish when removed from the cyst and fail to survive in sea water for more than 24 hours. The males, which have only been obtained when parasitic fifth copepodids molt in culture, are much smaller, averaging 0.8 mm, and are very active. Since one dead male has been found inside the cyst of an adult female and females are often found with attached spermatophores, it is suggested that the funnel of the cyst may also serve as an entrance for the males. Evidence is presented for the formation of the cyst as an accumulation of totipotent lymphocytes around the copepod. Cysts of parasitic developmental stages (third through fifth copepodids) are also described. All of these cysts and those of immature adult females lack funnels to the atrium. The funnel of the cyst of mature females is formed, in part, by modified cells of the wall of the blood vessel, but is induced after the major portion of the cellular cyst has been formed. Cells in the general circulation of the ascidian and those inside the lumen of the cyst are compared. The cells in the lumen of the mature cyst do not arise by diapedesis of blood cells from the subendostylar blood vessel, but by conversion and migration of cells composing the cyst proper. These cells have been found in the guts of the copepods and they may serve as a nutritive source. The ascidian appears not to be harmed by the association, but the copepod gains in many ways.     

Embryology ofEriocaulon setaceum (Eriocaulaceae)

Eriocaulon setaceum can be characterized by: young microsporangium wall with epidermis, endothecium (with fibrous thickenings), and glandular tapetum (uninucleate cells); pollen grains 3-celled, spiraperturate; embryo sac development according to the Polygonum type and with antipodal cyst; endosperm nuclear; embryo small, with incipient differentiation into cotyledonary and epicotylary loci; seed coat mainly from the inner layers of the integuments; pericarp 2-layered and membranous. Embryologically, theEriocaulaceae are nearer to theXyridaceae than to otherFarinosae. Their elevation to the rank of an order,Eriocaulales, therefore appears justified.     

Short-Cut Pathway to Synthesize Cellulose of Encysting Acanthamoeba

The mature cyst of Acanthamoeba is highly resistant to various antibiotics and therapeutic agents. Cyst wall of Acanthamoeba are composed of cellulose, acid-resistant proteins, lipids, and unidentified materials. Because cellulose is one of the primary components of the inner cyst wall, cellulose synthesis is essential to the process of cyst formation in Acanthamoeba. In this study, we hypothesized the key and short-step process in synthesis of cellulose from glycogen in encysting Acanthamoeba castellanii, and confirmed it by comparing the expression pattern of enzymes involving glycogenolysis and cellulose synthesis. The genes of 3 enzymes, glycogen phosphorylase, UDP-glucose pyrophosphorylase, and cellulose synthase, which are involved in the cellulose synthesis, were expressed high at the 1st and 2nd day of encystation. However, the phosphoglucomutase that facilitates the interconversion of glucose 1-phosphate and glucose 6-phosphate expressed low during encystation. This report identified the short-cut pathway of cellulose synthesis required for construction of the cyst wall during the encystation process in Acanthamoeba. This study provides important information to understand cyst wall formation in encysting Acanthamoeba.     

Effects of nitrogen deficiency and light of high intensity onCryptomonas rufescens (Cryptophyceae)

Summary C.rufescens excystment, experimentally induced, corresponds to a general metabolism recovery of the cell, previously in a resting phase. The cytoplasm changes without any polarity, and organelles like gullet and flagella redifferentiate. The thylakoids develop mainly from the stored lipidic compounds which then disappear. Phycoerythrin immediately fills the intrathylakoidal lumen. Pigment synthesis seems closely associated with the development of membranes. The activated cell divides and the cyst wall breaks down. The destruction of the wall begins in the median layer and is followed by a mechanical rupture of the external and internal layers. Each germinative cyst releases two or four fully differentiated cells. There is an exact symmetry between excystment and encystment, all the transformations of theC. rufescens cell being reversible.     

Cyst hatching in Anostraca accelerated by retinoic acid,amplified by Calcium Ionophore A23187, and inhibited by Calcium-channel blockers

Cyst hatching, under standardized conditions, of the Anostracan species Thamnocephalus platyurus and Streptocephalus dichotomus was significantly accelerated but not increased by applying the morphogen retinoic acid (RA). Cyst hatching was enhanced but not accelerated by artificially increasing the inflow of Ca²⁺ to the embryonic cells, using Calcium Ionophore A 23 187. Cyst hatching was accelerated and amplified, to a level in excess of the summed effects of each treatment, by a combined application of RA and ionophore. It was inhibited almost quantitatively by the Calcium-channel blockers Nifedipin and Verapamil. The significance of these findings is discussed.