Cyst Production Research Articles

Reconstruction of palaeoceanographic conditions in the South Atlantic Ocean at the last two Terminations based on calcareous dinoflagellate cysts

Despite the increasing interest in the South Atlantic Ocean as a key area of the heat exchange between the southern and the northern hemisphere, information about its palaeoceanographic conditions during transitions from glacial to interglacial stages, the so-called Terminations, are not well understood. Herein we attempt to increase this information by studying the calcareous dinoflagellate cysts and the shells of Thoracosphaera heimii (calcareous cysts) of five Late Quaternary South Atlantic Ocean cores. Extremely high accumulation rates of calcareous cysts at the Terminations might be due to a combined effect of increased cyst production and better preservation as result of calm, oligotrophic conditions in the upper water layers. Low relative abundance of Sphaerodinella albatrosiana compared with Sphaerodinella tuberosa in the Cape Basin may be the result of the relatively colder environmental conditions in this region compared with the equatorial Atlantic Ocean with high relative abundance of S. albatrosiana. Furthermore, the predominance of S. tuberosa during glacials and interglacials at the observed site of the western Atlantic Ocean reflects decreased salinity in the upper water layer.

Mechanisms underlying the formation and enlargement of noncommunicating syringomyelia: experimental studies

The pathogenesis of noncommunicating syringomyelia is unknown, and none of the existing theories adequately explains the production of cysts that occur in association with conditions other than Chiari malformation. The authors' hypothesis is that an arterial pulsation-driven perivascular flow of cerebrospinal fluid (CSF) is responsible for syrinx formation and enlargement. They investigated normal CSF flow patterns in 20 rats and five sheep by using the tracer horseradish peroxidase; the effect of reducing arterial pulse pressure was examined in four sheep by partially ligating the brachiocephalic trunk; CSF flow was examined in 78 rats with the intraparenchymal kaolin model of noncommunicating syringomyelia; and extracanalicular cysts were examined using the excitotoxic model in 38 rats. In the normal animals there was a rapid flow of CSF from the spinal subarachnoid space into the spinal cord perivascular spaces and then into the central canal. This flow ceased when arterial pulsations were diminished. In animals with noncommunicating syringomyelia, there was rapid CSF flow into isolated and enlarged segments of central canal, even when these cysts were causing pressure damage to the surrounding spinal cord. Exitotoxic injury of the spinal cord caused the formation of extracanalicular cysts, and larger cysts were produced when this injury was combined with arachnoiditis, which impaired subarachnoid CSF flow. The results of these experiments support the hypothesis that arterial pulsation-driven perivascular fluid flow is responsible for syrinx formation and enlargement.

Cyclin A Associates with the Fusome during Germline Cyst Formation in the Drosophila Ovary

Regulated changes in the cell cycle underlie many aspects of growth and differentiation. Prior to meiosis, germ cell cycles in many organisms become accelerated, synchronized, and modified to lack cytokinesis. These changes cause cysts of interconnected germ cells to form that typically contain 2n cells. InDrosophila, developing germ cells during this period contain a distinctive organelle, the fusome, that is required for normal cyst formation. We find that the cell cycle regulator Cyclin A transiently associates with the fusome during the cystocyte cell cycles, suggesting that fusome-associated Cyclin A drives the interconnected cells within each cyst synchronously into mitosis. In the presence of a normal fusome, overexpression of Cyclin A forces cysts through an extra round of cell division to produce cysts with 32 germline cells. Female sterile mutations in UbcD1, encoding an E2 ubiquitin-conjugating enzyme, have a similar effect. Our observations suggest that programmed changes in the expression and cytoplasmic localization of key cell cycle regulatory proteins control germline cyst production.

(Palaeo-)ecological significance, transport and preservation of organic-walled dinoflagellate cysts in the Somali Basin, NW Arabian Sea

To date, relatively little information is available about factors influencing organic-walled cyst production of tropical dinoflagellates and processes influencing the final burial of cysts in bottom sediments, such as transport and preservation. To extend this information, cyst fluxes were documented for three sediment traps from June 1992 to February 1993 at two sites in the Somali Basin (northwestern Arabian Sea) as well as the cyst association of underlying sediments. By comparing cyst associations of contemporaneously collected trap samples at different depths at one site, information about transport and processes of decay in the water column was obtained. Neither transport nor decay appears to have any detectable influence on cyst association during cyst settlement through the water column. Comparing the trap associations with the underlying sediments indicates that downslope transport appears to have influenced the cyst association on a local scale only. Species-selective decay, probably related to the presence of oxygen in bottom sediments, has influenced the cyst association most pronouncedly at the most offshore site. Relating variations in the trap associations with environmental conditions of the overlying surface waters indicates that highest production of both filled and empty cysts occurs during the SW Monsoon upwelling. Based on this correlation three groups of species can be distinguished: Species with highest fluxes during (1) the first-half of the SW Monsoon (June–August); Bitectatodinium spongium, Echinidinium granulatum, Echindinium transparantum, Echinidnium spp., cysts of Protoperidinium compressum and cysts of Protoperidinium subinerme, (2) the transition between the SW-Monsoon and inter-Monsoon; Spiniferites mirabilis and Spiniferites spp., (3) no particular season; all other species. Cyst associations of all trap samples are dominated by cyst of Protoperidinium species. Cysts with highest fluxes during the SW-Monsoon form about a third of the associations.

Apparent Global Ubiquity of Species in the Protist Genus Paraphysomonas

Evidence is presented for the ubiquity of protist species. Using the example of protists that leave traces (siliceous scales) of their recent population growth, we show that most - perhaps all species in the genus Paraphysomonas, are ubiquitous. Of the species recorded in surveys carried out worldwide, we have identified 78% of their number in 0.1 cm2 of sediment collected from a freshwater pond (total area 10(8) cm2) in England. Moreover, the pond appears to act like a microcosm of aquatic environments in general, for species that are globally rare or abundant, are likewise rare or abundant in the pond. We assume that the rate of neutral migration to the pond is greatest for the globally abundant species. As these species are probably capable of growth in a broad range of conditions, they will more frequently encounter the environment they require for population growth. Thus globally abundant species are also locally abundant in the pond - a pattern that will be amplified by periodic cyst production. Ubiquitous dispersal is probably driven by very high absolute abundance of individuals, and the water column of the pond was estimated to support >10(14) Paraphysomonas individuals. Ubiquity will dampen rates of speciation, and the evidence presented here indicates that global species richness of Paraphysomonas is indeed modest - perhaps close to what is already known.

Different excystment patterns in two calcareous cyst-producing species of the dinoflagellate genus Scrippsiella

Scrippsiella rotunda and Scrippsiella trochoidea var. aciculifera (order Peridiniales, subfamily Calciodinelloideae) are autotrophic orthoperidinioid dinoflagellates producing calcareous resting cysts, which are at times abundant in coastal marine sediments. We have carried out labora- tory experiments to investigate features of cyst germination in the two species, including dormancy length, germination pattern and germination success, over an annual cycle and under different light and temperature conditions. The maturation period for S.rotunda cysts was between 17 and 24 weeks, while that of S.trochoidea var. aciculifera was much shorter, ranging between 2 and 5 weeks. Both species required exposure to light for germination, while temperature shifts (from 14 to 20°C) in the dark did not induce excystment of mature cysts. In both species, germination was not synchronous, but distributed over a variable time interval, suggesting a high physiological diversity within the cyst pool. Moreover, exposure to light of S.rotunda cysts that had not completed maturation impaired the germination of a great percentage of the cysts. Differences in dormancy length may partially explain the distinct cyst production patterns observed for the two species in the Gulf of Naples.

Dinoflagellate cysts from sediment traps deployed in the Bellingshausen, Weddell and Scotia seas, Antarctica

Dinoflagellate cysts have been recovered from six long-term (1–2 yr) sediment trap moorings deployed in the Bellingshausen, Weddell and Scotia seas, Antarctica. These traps, mostly moored near the sea bed to sample the nepheloid layer, were located both within and to the north of the maximum sea-ice limit. The numbers of cysts, together with the composition of the assemblages, reinforce the importance of the maximum sea-ice limit as a modern biogeographic boundary for the distribution of dinoflagellate cysts. Cysts derived from heterotrophic dinoflagellates make up the highest proportions within the assemblages recovered from the traps. One trap sampled the export production, revealing little difference in cyst flux over those sampling the nepheloid layer. Cyst flux appears to be highest in areas closest to the Antarctic Convergence, north of the maximum sea-ice limit, and to high nutrient availability. There are, however, differences between the sediment trap assemblages and those recovered from core-top samples at the same or nearby sites. These differences, in the greater number of cysts, and in the higher numbers of round, brown Protoperidinium cysts in the traps, may reflect annual differences in the primary productivity and/or cyst production in the area. In some areas the sediment record may preserve little information about local surface water productivity because of the activity of bottom water currents, for example those arising from the Antarctic Circumpolar Current.

Congo Red Effect on Cyst Viability and Cell Wall Structure of Encysting Entamoeba invadens

Background The cell wall of Entamoeba invadens cysts is composed of chitin microfibrils as the main structural component. It has been demonstrated in yeast that the chitin cell wall assembly is altered by dyes such as Congo red (CR) and Calcofluor. Methods The purpose of this work was to study the cell wall assembly under the effect of CR dye on encysting E. invadens by means of light and electron microscopy, after the amebas were subjected to the effect of 100–2,000 μg CR/mL. Experiments were performed either in BI-S-33 or in mLG media. Results Trophozoite growth was not inhibited by 100–1,000 μg/mL CR after 8 days of incubation in BI-S-33 medium. However, low levels of growth were observed with 2,000 μg/mL of dye. No significant differences in morphologically viable (hyaline) cyst production occurred after 24–48 h, when 100 μg CR/mL was used, while the highest concentration of CR (2,000 μg/mL) resulted in a significant decrease of hyaline cyst yield; dead cysts prevailed in cultures, particularly at 72 h of CR treatment. Differentiation of amebas incubated in the presence of 500–2,000 μg/mL CR produced abnormal chitin deposits, rendering irregularly thick or double cell walls, as shown by transmission and scanning electron microscopy. Cyst cultures obtained under 100 μg/mL CR produced as many trophozoites as did the control when they were incubated in BI-S-33, but only low numbers of trophozoites were found in culture cysts obtained under higher CR doses. Conclusion Our results suggest that CR affects E. invadens encystment, alters the cell wall formation, and also affects the cyst viability.

International study on Artemia. LX. Allozyme data suggest that a new Artemia population in southern Chile (50° 29′ S; 73° 45′ W) is A. persimilis

An Artemia population from a site recently described in Chile and located in a semi-arid and humid hydrographical zone (Torres del Paine National Park), was electrophoretically characterized and cross-bred (followed through F 3) to reference samples of A. franciscana (San Francisco Bay, California, U.S.A.) and A. persimilis (Buenos Aires, Argentina), which are the species most likely to be found in Chile. The population exhibited high levels of genetic variability as judged by the percentage of polymorphic loci (P=53.3), mean number of alleles per locus (n=2.2) and mean heterozygosity ( $$\bar H$$ =0.176). Based on the analysis of 15 loci it seems to be closer to A. persimilis (D Nei=0.211) than to A. franciscana (D=0.591). Laboratory cross-fertility tests do not suggest barriers to gene flow or infertility in accordance with eletrophoretic results (genetic distance values or population-specific alleles), though offspring quality tended to be biased toward the production of cysts rather than nauplii, most obviously in F33, which may be an indication of post-zygotic reproductive isolation.

Decoquinate induces tissue cyst formation by the RH strain of Toxoplasma gondii

Decoquinate is an anticoccidial agent that inhibits respiration in the parasites mitochondrion. We examined human foreskin fibroblast cell cultures infected with the normally tissue cyst-less RH strain of Toxoplasma gondii and treated with decoquinate for evidence of tissue cyst induction and formation. Transmission electron microscopy observations demonstrated tissue cysts in decoquinate-treated cultures on days 3, 4, 5, and 6 after inoculation. Tissue cysts contained a tissue cyst wall that enclosed stages that resembled tachyzoites and stages that were structurally bradyzoites. Similar treatment of human foreskin fibroblast cells infected with tachyzoites of the TS-4 temperature-sensitive mutant of the RH strain did not result in production of tissue cysts.

Cysts of Danish Gymnodinium nolleri Ellegaard et Moestrup sp. ined. (Dinophyceae): studies on encystment, excystment and toxicity

Life cycle dynamics of Gymnodinium nolleri Ellegaard et Moestrup sp. ined. were studied under different temperature and nutrient conditions. Five culture strains originating from cysts found in Danish marine sediments were used for the experiments. Both encystment and excystment were found to vary with temperature. Maximal encystment occurred at 22-28°C, with no cysts formed below 13°C or above 33°C. Cyst production was slightly higher under phosphorus limitation than under combined nitrogen and phosphorus limitation. Maximal excystment occurred at 26.5°C with negligible excystment under 11°C and over 35°C. The resting period for cyst maturation was typically 3-4 weeks. Cysts produced under both phosphorus and nitrogen limitation were tested for paralytic shellfish poisoning (PSP) toxins by HPLC, and none were detected. It remains unclear why vegeta- tive cells of this species have not yet been recorded in plankton samples from Scandinavia, despite the widespread distribution of the cysts.

Dinoflagellate cyst production at a coastal Mediterranean site

Dinoflagellate cyst production at a coastal Mediterranean site

Artemia parthenogenetica in Lake Hayward, Western Australia. I. Interrupted recruitment into adult stages in response to seasonal limnology

Lake Hayward, Western Australia, is a pristine, monomictic coastal lake with a residual brine (depth=1.5 m) of 200 gL−1: stratification in winter (May to October) followed the input of ground water and rain. The epilimnion had temperature ranges of 10–30°C, and salinity from 65–110 g L−1. Maximum depth in winter was 2.4 m. The population structure of the brine shrimpArtemia parthenogenetica, the dominant metazoan grazer in Lake Hayward, was studied from September, 1989, to February, 1991. The L. Hayward population ofA. parthenogenetica is predominantly ovoviviparous with little recruitment into the adult population. Cyst production occurs in summer, and there was evidence for recruitment from cyst hatching in the winters of 1989 and 1990. Nauplii were continuously present throughout the study period. Nevertheless, recruitment through to adult stages as defined by the presence of gravid females occurred only twice: in October–December, 1989 and in December, 1990. Based on limited observations of long-term survival of laboratory cultures, we believe that the major mechanism controlling nauplius survival and recruitment ofArtemia in Lake Hayward is food quality and quantity.

A Protocol for the Production of Neospora caninum Tissue Cysts in Mice

Identification of a definitive host for Neospora caninum has been inhibited by lack of an efficient method for producing bradyzoites, needed for oral infectivity trials. An improved protocol for producing bradyzoite-containing tissue cysts in mouse brains is described. Six variables, including mouse strain (Balb/C, CBA/Ca, and ICR), sex, N. caninum isolate (NC-2 and NC-Liverpool), tachyzoite inoculum dose, immunosuppression with methylprednisolone acetate (MPA), and sulfadiazine treatment were tested. Tissue cyst numbers were estimated using an immunohistologic staining procedure specific for bradyzoites. Male ICR mice (> or = 30 g) that were immunosuppressed with 2 mg MPA 7 days prior to and 2.5 mg MPA at the time of subcutaneous inoculation with 400,000 N. caninum tachyzoites produced the highest numbers of tissue cysts. Significant numbers were produced by methods using the NC-2 strain of N. caninum; however, protocols using NC-Liverpool produced greater numbers of tissue cysts. Sulfadiazine treatment did not appear to contribute to tissue cyst production. The procedure described is superior to previously described methods with regard to numbers of tissue cysts produced, protocol reproducibility, and survival of mice until tissue cyst formation.

Preservation of organic-walled dinoflagellate cysts in different oxygen regimes: a 10,000 year natural experiment

The occurrence of organic-walled dinoflagellate cysts in (fossil) sediments depends on several factors, including as the ecological preferences of the cyst-forming dinoflagellates, cyst production, transport and preservation. Although laboratory experiments have shown that several cyst species are sensitive to chemical treatment, no information about the selective preservation of dinoflagellate cyst species in natural environments has previously been presented. Here, we present data on the effects of oxygen availability in bottom sediments on a cyst assemblage from the ungraded Madeira Abyssal Plain f-turbidite of which only the upper layer has been oxidized. Based on differences in species composition between the oxidized and underlying, unoxidized layers of this turbidite, the influence of oxygen availability on the preservation of individual species has been estimated. Cyst species have been classified in ascending order of resistance to oxygen availability in sediments as: (1) highly sensitive (cysts formed by Protoperidinium species), (2) moderately sensitive (e.g. Spiniferites species), (3) moderately resistant (e.g. Impagidinium paradoxum and Nematosphaeropsis labyrinthus) and (4) resistant (e.g. Impagidinium aculeatum).

Developmental changes in the tegument of four microphallid metacercariae in their second (crustacean) intermediate hosts

AbstractThe morphology of the tegument of four microphallid metacercariae from the stage of invasive cercariae to their maturation as encysted metacercariae inside their crustacean second intermediate hosts is described. The tegument of the metacercariae developed surface lamellae and projections which, along with coated vesicles in the surface syncytium, indicated that the tegument had an absorptive function. The disappearance of secretory granules from the tegument at the same time as the appearance of the first cyst wall suggested that the tegument had a role in primary cyst production. Following this, the metacercariae continued to grow and seemingly retained their absorptive ability. The tegument was also involved in the transport of material into the perimetacercarial lumen prior to its eventual inclusion in the developing inner cyst layers. It appeared that this material originated in tegumental cells located amongst the parenchymal cells beneath the tegumental syncytial layer. On completion of the secondary cyst layers there was a gradual degeneration of structures associated with absorption and a progressive accumulation of dense discoid granules traceable to underlying tegumental cells. All four microphallid species (Maritrema arenaria,M. subdolum,Levinseniella brachysomaandMicrophallus claviformis) demonstrated the same developmental pattern but the period spent in each stage differed depending on the time spent migrating to encystment sites. The pattern of tegumental development described is thought to be applicable to all microphallid metacercariae and possibly to other metacercariae which undergo growth and development in their second intermediate hosts.

A ubiquitous intracellular parasite: The cellular biology of Toxoplasma gondii

Toxoplasma gondii shares many features with other apicomplexan parasites but is unusual in its extremely broad host and tissue specificity. The parasite exhibits typical ‘zoite’ morphology, its highly polar structure being dictated by the complex cytoskeleton. Molecules on the surface of the zoite are prime candidates for interaction with the host cell and in vitro assays have implicated 2 of the 5 tachyzoite surface molecules in invasion: SAG1 as a ligand mediating host cell invasion, and SAG2 in enabling reorientation prior to invasion. The functional roles of other molecules, secreted from internal organelles during invasion and intracellular development, are also becoming clear through immuno-EM and biochemical studies, and from sequence data. Molecules from the rhoptries including the penetration enhancing factor ROP1 are secreted at the point of invasion and are integral to the newly formed parasitophorous vacuole membrane. Release of the dense granule molecules GRA 1–6, appears to be calcium regulated and occurs within 10 min of invasion leading to formation of the tubular membranous network and stabilization of the vacuole. The interaction between Toxoplasma and the host cell is stage specific. The tachyzoite divides rapidly and synchronously forming rosettes and causing host cell lysis, while the bradyzoite exhibits slow asynchronous division secreting a granular matrix and becoming enclosed within a cyst wall. This altered phenotype is a reflection of changes in gene expression. Bradyzoite specific molecules are found internally, on the parasite surface, and in the cyst matrix while important tachyzoite proteins such as SAG1 and SAG2 are downregulated. Differentiation between the 2 stages is reversible and is influenced by immunomodulatory agents. However a strong genetic element is involved and it is notable that virulent strains show a very low frequency of cyst production.

Variations in some physico-chemical parameters in a hypersaline coastal lagoon of Baja California, Mexico

San Jose lagoon is a hypersaline body of water located in Mexico in the Baja California Peninsula. The lagoon belongs to a system that lies between the fault ridge known as San Jose Creek. Because of its marine origin, it can be considered as thalassohaline, but its isolation from the ocean has brought about changes in its salt composition. It has an area of 13,500 m2, a mean depth of 80 cm and a total volume of 10,000 m3. It does not desiccate and can be considered as a permanent lagoon. Seasonal variations are small. TheArtemia population in San Jose produces cysts all year. To determine the physico-chemical conditions inducing permanent production of cysts, temperature, salinity, dissolved oxygen, and pH of the lagoon were monitored, as well as relative humidity and wind conditions in the region in different seasons of the year. From spring to summer, differences of 1 mg L−1 of O2, 1°C in water temperature, and 8 g L−1 in salinity were observed, and from summer to winter differences of 3.3 mg L−1, 6.5°C, and 14 g L−1, respectively. Despite small seasonal variations, the lagoon exhibits strong spatial and daily changes that are important for cyst production.

Relative susceptibilities of eleven potato cultivars and breeders' clones toGlobodera pallida pathotype Pa 3, with a discussion of the interpretation of data from pot experiments

Curves according to the equationPf=M′ (1-e −aPi/M′ ) fitted well to the relations according to Seinhorst's (1993) modelPf=y e y h M(1-e −aPi/M ) between initial egg densities of potato cyst nematodes (Globodera rostochiensis, G. pallida)Pi up to 5T h (T h =the tolerance limit of haulm weight) andPf at the end of the growing season (a=maximum rate of reproduction,M, M′=different theoretical maximum egg densities). Variation of estimates ofM′, due to variation of the parameters of the submodelsy e andy h for the effect of weight reduction of haulms (and, therefore, of roots) on cyst production and damage to root tissue on egg production, respectively, was small enough to be ignored relative to experimental error. Therefore, ratios of values ofM′, determined in simultaneous pot experiments with different potato cultivars, are reliable measures of the relative host status of these cultivars at initial egg densitiesPi of these nematodes up to about 5T h . Variation between potato cultivars of growth reduction and damage to root tissue by the nematodes reduces the reliability of ratios of rates of relative susceptibility of these cultivars. The ratios between the maximum rates of reproduction ofG. pallida, pathotype Pa 3, on 8 out of 9 cultivars and one breeder's clone of potatoes and this rate on the susceptible cvs Bintje and Irene (relative susceptibilitiesrs a ) could be considered to be equal to the ratios of maximum population densitiesM′ on these cultivars (relative susceptibilitiesrs M′ ) (relative susceptibilities independent of initial egg density). The latter ratios were 0.59 times the first (relative susceptibilities negatively correlated with initial egg density) in one cultivar and one breeder's clone. Relative susceptibilitiesrs a andrs M′ of the tested cultivars and breeders clones suggest the existence of continuous ranges of both relative susceptibilities between 0.50 and 0.15 with, in a great majority of cases,rs a =rs M′ .

Field Population Dynamics of Soybean Cyst Nematode on Resistant and Susceptible Soybeans and Their Blends

Soybean cyst nematode (Heterodera glycines Ichinohe) (SCN) become an increasing problem in the upper midwest. In order to develop appropriate practices to manage SCN populations it is important to understand the population dynamics of SCN under field conditions. Soybean cyst nematode population dynamics were determined in naturally infested commercial soybean [Glycine max (L.) Merr.] fields in southern Minnesota on a monthly basis during 1991 and 1992 for an adapted susceptible (S) and a SCN Race 3 resistant (R) cultivar and three blends (25% R:75% S; 50% R:50% S; 75% R: 25% S). Population dynamics differed between sites and years. In 1991, fields with similar initial cyst densities were compared. Densities of second‐stage juveniles (J2) peaked in August at one site and gradually declined throughout the growing season at the other. Total cyst production was lower at the site where J2 densities declined. In 1992, fields with high and low initial egg densities were compared. SCN population densities increased when initial densities of J2 and eggs were low and decreased when initial J2 and egg densities were high. Physiological race of the nematode influenced population dynamics on resistant plants. The cyst index under field conditions for the resistant cultivar was 10 (moderately resistant) against Race and 32 (moderately susceptible) against Race 1. The blends generally lowered SCN population densities, but not significantly. Final egg density and cyst production for the resistant cultivar were significantly lower than the susceptible cultivar at each site. The use of SCN resistant cultivars alone or in a 75:25% mixture with a susceptible cultivar (25%) is recommended as a SCN population management strategy to delay shifts from one race to another thus prolonging the effectiveness of resistant cultivars.