Dormant Cysts Research Articles

MALE-FEMALE COEVOLUTION IN THE WILD: EVIDENCE FROM A TIME SERIES IN ARTEMIA FRANCISCANA

Sexual conflicts are ubiquitous in nature and are expected to lead to an antagonistic coevolution between the sexes. This coevolutionary process is driven by selection on sexually antagonistic traits that can either be directional or fluctuating. In this study, we used dormant cysts of Artemia franciscana, collected in the same population in three different years over a 23-year period (corresponding to ∼160 generations in this system), to investigate male-female coevolution in natural conditions over time. We performed a cross experiment study where reproduction of females mated to males from the past, present, or future was monitored until death. In agreement with a model of "fluctuating selection," we found that females survived better and had longer interbrood intervals when mated with their contemporary males compared to when mated with males from the future or the past. However, female weekly and lifetime reproductive successes displayed no differences between contemporary and noncontemporary matings. Finally, the coevolutionary patterns ("arms race dynamics" or "fluctuating selection dynamics") possibly acting on female relative fitness could not be discriminated. This study is the first direct demonstration that the process of male-female coevolution, previously revealed by experimental evolution in laboratory artificial conditions, can occur in nature on a short evolutionary time scale.

A Novel E2F-like Protein Involved in Transcriptional Activation of Cyst Wall Protein Genes in Giardia lamblia

Giardia lamblia differentiates into resistant walled cysts for survival outside the host and transmission. During encystation, synthesis of cyst wall proteins is coordinately induced. The E2F family of transcription factors in higher eukaryotes is involved in cell cycle progression and cell differentiation. We asked whether Giardia has E2F-like genes and whether they influence gene expression during Giardia encystation. Blast searches of the Giardia genome database identified one gene (e2f1) encoding a putative E2F protein with two putative DNA-binding domains. We found that the e2f1 gene expression levels increased significantly during encystation. Epitope-tagged E2F1 was found to localize to nuclei. Recombinant E2F1 specifically bound to the thymidine kinase and cwp1-3 gene promoters. E2F1 contains several key residues for DNA binding, and mutation analysis revealed that its binding sequence is similar to those of the known E2F family proteins. The E2F1-binding sequences were positive cis-acting elements of the thymidine kinase and cwp1 promoters. We also found that E2F1 transactivated the thymidine kinase and cwp1 promoters through its binding sequences in vivo. Interestingly, E2F1 overexpression resulted in a significant increase of the levels of CWP1 protein, cwp1-3 gene mRNA, and cyst formation. We also found E2F1 can interact with Myb2, a transcription factor that coordinate up-regulates the cwp1-3 genes during encystation. Our results suggest that E2F family has been conserved during evolution and that E2F1 is an important transcription factor in regulation of the Giardia cwp genes, which are key to Giardia differentiation into cysts.

Trehalose and Vitreous States: Desiccation Tolerance of Dormant Stages of the CrustaceansTriopsandDaphnia

Several aquatic organisms are able to withstand extreme desiccation in at least one of their life stages. This is commonly known as "anhydrobiosis." It was often thought that to tolerate such a desiccated state required high amounts of compatible solutes such as the nonreducing disaccharide trehalose, which protects cellular structures by water replacement and glass formation. Trehalose levels of dormant eggs and cysts of five freshwater crustaceans (Daphnia magna, Daphnia pulex, Triops longicaudatus, Triops cancriformis, and Triops australiensis) were observed in different states of hydration and dehydration. Although trehalose was detected in all species, the concentration was under 0.5% of the dry weight (0.05 μg/μg protein), and no change between the different states was observed. Differential scanning calorimetry (DSC) measurements indicated that dried cysts of all Triops species were in a glassy state, supporting the vitrification hypothesis. No indication for a vitreous state was found in dried resting eggs of Daphnia.

Ecosystem-level effects of bioturbation by the tadpole shrimp Lepidurus packardi in temporary pond mesocosms

An example of ecosystem engineering gaining attention in aquatic systems is bioturbation, the disruption of sediment at the water–sediment interface due to burrowing and foraging. One consequence of bioturbation can be increased turbidity from suspended sediment, which generally inhibits macrophyte growth and reduces ecosystem functioning. Conversely, bioturbation may promote invertebrate species richness by unearthing dormant cysts. Temporary-pond crustaceans are not widely regarded as agents of bioturbation, but on the basis of aquaria observations we hypothesized that certain taxa can disturb the sediment and create highly turbid water. We tested this hypothesis by removing crustaceans from mesocosms lined with vernal pool soil. Compared to this treatment group, mesocosms containing crustaceans had extremely high turbidity from suspended sediment, as well as reduced total macrophyte cover. We also found clear compositional differences in macrophyte communities between treatments, driven largely by differences in water physicochemistry, including turbidity. Regression analysis linked most of the bioturbation to the endangered notostracan Lepiduruspackardi Simon 1886, which was a strong predictor of turbidity in our mesocosms. We also found a trend toward increased crustacean species richness in our mesocosms in the presence of this taxon. An analysis of published data from King et al. (1996) suggests that this trend may extend to natural vernal pools. Overall, our results suggest that L. packardi may have large effects on vernal pool communities, likely mediated in part through its disturbing of the sediment.

A comparison of the Giardia lamblia trophozoite and cyst transcriptome using microarrays

BackgroundCompared with many protists, Giardia lamblia has a simple life cycle alternating between cyst and trophozoite. Most research on the molecular biology of Giardia parasites has focused on trophozoites and the processes of excystation and encystation, whereas cysts have attracted less interest. The striking morphological differences between the dormant cyst and the rapidly dividing and motile trophozoite implies profound changes in the metabolism as the parasite encysts in the host's intestine and excysts upon ingestion by a new host.ResultsTo investigate the magnitude of the transcriptional changes occurring during the G. lamblia life cycle we compared the transcriptome of G. lamblia trophozoites and cysts using single-color oligonucleotide microarrays. Cysts were found to possess a much smaller transcriptome, both in terms of mRNA diversity and abundance. Genes encoding proteins related to ribosomal functions are highly over-represented. The comparison of the transcriptome of cysts generated in culture or extracted from feces revealed little overlap, raising the possibility of significant biological differences between the two types of cysts.ConclusionsThe comparison of the G. lamblia cyst and trophozoite transcriptome showed that transcripts of most genes are present at a lower level in cysts. This global view of the cyst and trophozoite transcriptome complements studies focused on the expression of selected genes during trophozoite multiplication, encystation and excystation.

Two putative histidine kinases are required for cyst formation in Rhodospirillum Centenum

The photosynthetic bacterium, Rhodospirillum centenum, has a flexible life cycle that permits it to survive starvation as dormant cyst cells. Previous studies have identified some of the key regulators for encystment and demonstrated that the control of development is intricate. This complexity may arise from the need to integrate several environmental signals to mediate a switch from one mode of energy metabolism to another and to ensure that a transition to dormancy is initiated only when necessary. We searched for additional regulators of development by screening for encystment deficient strains after subjecting wild type R. centenum to mini-Tn5 mutagenesis. Analysis of "hypo-cyst" strains led to the identification of two genes that encode putative hybrid histidine kinases (cyd1 and cyd2). Cells with deletions of either gene fail to form cysts under conditions that normally induce development. Furthermore, the deletion strains exhibit altered swarming behavior suggesting that Cyd1 and Cyd2 affect behaviors utilized when the organism is attached to a substrate.

Toxoplasma gondii in the peripheral blood of patients with acute and chronic toxoplasmosis

Background and aimsToxoplasmic retinochoroiditis may recur months or years after the primary infection. Rupture of dormant cysts in the retina is the accepted hypothesis to explain recurrence. Here, the authors...

Major Role for Cysteine Proteases during the Early Phase of Acanthamoeba castellanii Encystment

Acanthamoeba castellanii is a facultative pathogen that has a two-stage life cycle comprising the vegetatively growing trophozoite stage and the dormant cyst stage. Cysts are formed when the cell encounters unfavorable conditions, such as environmental stress or food deprivation. Due to their rigid double-layered wall, Acanthamoeba cysts are highly resistant to antiamoebic drugs. This is problematic as cysts can survive initially successful chemotherapeutic treatment and cause relapse of the disease. We studied the Acanthamoeba encystment process by using two-dimensional gel electrophoresis (2DE) and found that most changes in the protein content occur early in the process. Truncated actin isoforms were found to abound in the encysting cell, and the levels of translation elongation factor 2 (EF2) were sharply decreased, indicating that the rate of protein synthesis must be low at this stage. In the advanced stage of encystment, however, EF2 levels and the trophozoite proteome were partly restored. The protease inhibitors PMSF (phenylmethylsulfonyl fluoride) and E64d [(2S,3S)-trans-epoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester] inhibited the onset of encystment, whereas the protein synthesis inhibitor cycloheximide was ineffective. Changes in the protein profile, similar to those of encysting cells, could be observed with trophozoite homogenates incubated at room temperature for several hours. Interestingly, these changes could be inhibited significantly by cysteine protease inhibitors but not by inhibitors against other proteases. Taken together, we conclude that the encystment process in A. castellanii is of a bipartite nature consisting of an initial phase of autolysis and protein degradation and an advanced stage of restoration accompanied by the expression of encystment-specific genes.

Stress-driven stage transformation of Neospora caninum

Neosporiosis, a serious disease caused by the apicomplexan intracellular parasite Neospora caninum, is considered to be one of the most economically important diseases in cattle. It is associated with potentially serious complications such as abortion, stillbirth and maternal infertility. To survive in fluctuating physiological and immunological environments, N. caninum has evolved a diverse set of regulatory mechanisms that govern various adaptive responses. The most intriguing paradigm in N. caninum adaptive evolutionary biology is its ability to alternate between two phenotypically and functionally distinct stages within the host. Recent research has reinforced the notion that N. caninum tachyzoite-bradyzoite stage switching on and off is correlated with its ability to form dormant cysts. Knowledge of the specific mechanisms that govern the dynamics of N. caninum phenotype switching enables a better understanding of the pathogenesis of the disease and effective control measures to be identified. Herein we review the available knowledge relating to various aspects of stage interconversion in N. caninum, with particular focus on the stress-related hypothesis presumed to be involved in this event. Finally, we put forward the postulation that N. caninum uses stage interconversion as an adaptive process to cope with the hostile environment within the host and to ensure its continuity in nature.

Origin of the cell nucleus, mitosis and sex: roles of intracellular coevolution.

BackgroundThe transition from prokaryotes to eukaryotes was the most radical change in cell organisation since life began, with the largest ever burst of gene duplication and novelty. According to the coevolutionary theory of eukaryote origins, the fundamental innovations were the concerted origins of the endomembrane system and cytoskeleton, subsequently recruited to form the cell nucleus and coevolving mitotic apparatus, with numerous genetic eukaryotic novelties inevitable consequences of this compartmentation and novel DNA segregation mechanism. Physical and mutational mechanisms of origin of the nucleus are seldom considered beyond the long-standing assumption that it involved wrapping pre-existing endomembranes around chromatin. Discussions on the origin of sex typically overlook its association with protozoan entry into dormant walled cysts and the likely simultaneous coevolutionary, not sequential, origin of mitosis and meiosis.ResultsI elucidate nuclear and mitotic coevolution, explaining the origins of dicer and small centromeric RNAs for positionally controlling centromeric heterochromatin, and how 27 major features of the cell nucleus evolved in four logical stages, making both mechanisms and selective advantages explicit: two initial stages (origin of 30 nm chromatin fibres, enabling DNA compaction; and firmer attachment of endomembranes to heterochromatin) protected DNA and nascent RNA from shearing by novel molecular motors mediating vesicle transport, division, and cytoplasmic motility. Then octagonal nuclear pore complexes (NPCs) arguably evolved from COPII coated vesicle proteins trapped in clumps by Ran GTPase-mediated cisternal fusion that generated the fenestrated nuclear envelope, preventing lethal complete cisternal fusion, and allowing passive protein and RNA exchange. Finally, plugging NPC lumens by an FG-nucleoporin meshwork and adopting karyopherins for nucleocytoplasmic exchange conferred compartmentation advantages. These successive changes took place in naked growing cells, probably as indirect consequences of the origin of phagotrophy. The first eukaryote had 1-2 cilia and also walled resting cysts; I outline how encystation may have promoted the origin of meiotic sex. I also explain why many alternative ideas are inadequate.ConclusionNuclear pore complexes are evolutionary chimaeras of endomembrane- and mitosis-related chromatin-associated proteins. The keys to understanding eukaryogenesis are a proper phylogenetic context and understanding organelle coevolution: how innovations in one cell component caused repercussions on others.ReviewersThis article was reviewed by Anthony Poole, Gáspár Jékely and Eugene Koonin.

Two red eyes and one asymptomatic donor

In October, 2008, a 30-year-old man underwent penetrating keratoplasty (PK) for keratoconus. A week later, he presented with ocular pain, ciliary injection, and graft oedema, with keratic precipitates. Despite treatment with antibiotics and steroids there was no improvement, and he underwent a re-grafting procedure in November. Analysis of the excised corneal button showed the presence of acanthamoeba cysts deeply embedded within the stroma (fi gure A). We treated him with topical hexamidine and 0·02% polihexanide. Keratitis recurred and a third PK was done in February, 2009, after the second graft melted. When last seen, in June, 2009, the patient had epithelial defects of the cornea but no signs of acanthamoeba infection; visual acuity was up to counting fi ngers. Also in October, 2008, the fellow cornea from the same donor was transplanted into a 73-year-old woman with post-traumatic corneal leukoma. Con comitant vitrectomy, extracapsular cataract extraction, and intraocular lens insertion were done. Despite treatment with oral and topical antibiotics, a worsening course (fi gure B) led to a second PK in December, 2008. Again, the explanted graft showed stromal acanthamoeba cysts. She was treated with aminoglycoside, 0·02% polihexanide and 0·1% propamidine eyedrops. When last seen, in June, 2009, she showed phthisis bulbi with no light perception. PCR of the 18S ribosomal subunit was done on DNA from both explanted grafts with JDP1 and JDP2 primers. PCR bands of 440 bp were sequenced and BLASTed (Basic Local Alignment Search Tool); they showed 98% homology to Acanthamoeba sp CRIB-20 and Acanthamoeba castellanii 18S sequences. Sequences of PCR bands from the two explanted corneas were identical, suggesting a single source of infection. The transplanted corneas were from a 45-year-old male donor who died after a motorcycle accident in September, 2008. He had never worn contact lenses nor complained of ocular discomfort or redness. He had been wearing his helmet when pronounced dead and his eyes were not exposed to sources of contamination until the corneas were retrieved 26 h later. Slit lamp and light microscopy confi rmed that both corneas were suitable for transplantation. No stromal abnormalities were seen. Routine microbiological tests were negative. We concluded that donor-to-recipient transmission of acanthamoeba from an asymptomatic individual had occurred, with dormant cysts residing in the donor corneas without active infection or morbidity. Acanthamoeba are ubiquitous protozoa in soil, water, and air. Risk factors for infection include use of contact lenses, corneal trauma, and contaminated fl uids. In the USA and the Netherlands, respectively, 1–2 and 3 people per million contact lens wearers are aff ected. Acanthamoeba keratitis can progress to scleritis, and uncontrolled infections require enucleation. Misdiagnosis as herpes or fungal keratitis often delays treatment. It is unlikely that a simultaneous ocular infection occurred in our two patients or that they had been at risk before transplantation (rigid gas-permeable contact lenses were negative for acanthamoeba culture in the fi rst patient; trauma had occurred 40 years earlier in the second patient). Simul taneous contamination of donor tissues in the eye bank was also excluded, because other corneas processed on the same days were transplanted uneventfully. A few cases of painless acanthamoeba keratitis have been reported, also in patients not wearing contact lenses. Since the presence of acanthamoeba in corneas of asymptomatic donors is unlikely, the cost/benefi t ratio of screening for acanth amoeba before transplantation is low. However, because the consequences of donormediated acanth amoeba infec tions are deleterious, eye banks should be aware of these rare events, develop a screening test, and work closely with surgeons to diagnose and treat keratitis rapidly.

Reciprocal Expression of Integration Host Factor and HU in the Developmental Cycle and Infectivity of Legionella pneumophila

Legionella pneumophila is an intracellular parasite of protozoa that differentiates late in infection into metabolically dormant cysts that are highly infectious. Regulation of this process is poorly understood. Here we report that the small DNA binding regulatory proteins integration host factor (IHF) and HU are reciprocally expressed over the developmental cycle, with HU expressed during exponential phase and IHF expressed postexponentially. To assess the role of these regulatory proteins in development, chromosomal deletions were constructed. Single (ihfA or ihfB) and double deletion (Deltaihf) IHF mutants failed to grow in Acanthamoeba castellanii unless complemented in trans when expressed temporally from the ihfA promoter but not under P(tac) (isopropyl-beta-d-thiogalactopyranoside). In contrast, IHF mutants were infectious for HeLa cells, though electron microscopic examination revealed defects in late-stage cyst morphogenesis (thickened cell wall, intracytoplasmic membranes, and inclusions of poly-beta-hydroxybutyrate), and were depressed for the developmental marker MagA. Green fluorescent protein promoter fusion assays indicated that IHF and the stationary-phase sigma factor RpoS were required for full postexponential expression of magA. Finally, defects in cyst morphogenesis noted for Deltaihf mutants in HeLa cells correlated with a loss of both detergent resistance and hyperinfectivity compared with results for wild-type cysts. These studies establish IHF and HU as markers of developmental stages and show that IHF function is required for both differentiation and full virulence of L. pneumophila in natural amoebic hosts.

The ecology and biological affinity of Arctic dinoflagellates and their paleoceanographical significance in the Canadian High Arctic

Dinoflagellates are eukaryotic organisms and constitute an important group of marine primary producers. Approximately 10-15% of living dinoflagellates produce a highly resistant dormant cyst that is fossilisable, and which constitute an excellent proxy indicator of the upper water column conditions and productivity. Relatively little is known on the distribution in time and space of the dinoflagellate life cycle (i.e., vegetative and cyst stages) in the Canadian Arctic; most studies usually focusing on other groups of organisms (e.g., diatoms). Here, we present information on the ecology of dinoflagellate cysts and how they relate to their counterpart plankton stages. We discuss the importance of considering the biological affinities of dinoflagellates cysts and their relevance for paleoceanographical interpretations. We also provide insight on the actual lack of such knowledge for the Canadian Arctic cyst and plankton assemblages.

Stress-induced tRNA-derived RNAs: a novel class of small RNAs in the primitive eukaryote Giardia lamblia

Giardia lamblia is an early diverging and evolutionarily successful protozoan as it can enter into a dormant cyst stage from a vegetative trophozoite. During dormant stage, its metabolic rate decreases dramatically. However, to date, the regulatory molecules participating in the initiation and maintenance of this process have not been fully investigated. In this study, we have identified a class of abundant small RNAs named sitRNAs, which are ∼46 nucleotides in length and accumulate in G. lamblia encysting cultures. Remarkably, they are derived from the 3′ portion of fully matured tRNAs by cleavage of the anticodon left arm, with the 3′ terminal CCA triplex still connected. During differentiation, only a limited portion of mature tRNAs is cleaved, but this cleavage occurs almost in the entire tRNA family. sitRNAs begin to accumulate as early as 3 h after initiation of encystation and are maintained at a relatively stable level during the whole process, exhibiting an expression peak at around 24 hr. Our studies further show that sitRNAs can be induced by several other stress factors, and in the case of serum deprivation, both tRNAs and sitRNAs degrade rapidly, with the accumulation of tRNA being halved. Our results may provide new insight into a novel mechanism for stressed G. lamblia to regulate gene expression globally.

Three-dimensional structure of the ciliate Didinium nasutum nucleoli

The nucleolar organization in ciliate Didinium nasutum somatic interphase nuclei was studied using serial ultrathin sections and compared for various physiological states of the cell, namely, fed ciliates, starved ciliates, and dormant cysts. It has been shown that the interphase nucleoli are large structures with a complex architecture: the fibrillar component forms an intricate network in the macronucleus space, while the granular component is located inside this network. The structures looking as individual nucleoli in single sections are actually parts of branched nucleolar networks. The intricate nucleolar networks do not disintegrate after a 30-h starvation; however, the granular component becomes denser and develops numerous cavities filled with fine fibrils of a nonribonucleoprotein nature. In fed D. nasutum, the fibrillar structures on the periphery of nucleoli contain numerous pores (virtually absent in starved cell nucleoli), which can potentially serve for transporting newly synthesized rRNP. Branched nucleolar networks are undetectable in cysts. Their nucleoli are individual structures consisting mainly of the fibrogranular component.

Direct transfer of starter substrates from type I fatty acid synthase to type III polyketide synthases in phenolic lipid synthesis

Alkylresorcinols and alkylpyrones, which have a polar aromatic ring and a hydrophobic alkyl chain, are phenolic lipids found in plants, fungi, and bacteria. In the Gram-negative bacterium Azotobacter vinelandii, phenolic lipids in the membrane of dormant cysts are essential for encystment. The aromatic moieties of the phenolic lipids in A. vinelandii are synthesized by two type III polyketide synthases (PKSs), ArsB and ArsC, which are encoded by the ars operon. However, details of the synthesis of hydrophobic acyl chains, which might serve as starter substrates for the type III polyketide synthases (PKSs), were unknown. Here, we show that two type I fatty acid synthases (FASs), ArsA and ArsD, which are members of the ars operon, are responsible for the biosynthesis of C(22)-C(26) fatty acids from malonyl-CoA. In vivo and in vitro reconstitution of phenolic lipid synthesis systems with the Ars enzymes suggested that the C(22)-C(26) fatty acids produced by ArsA and ArsD remained attached to the ACP domain of ArsA and were transferred hand-to-hand to the active-site cysteine residues of ArsB and ArsC. The type III PKSs then used the fatty acids as starter substrates and carried out two or three extensions with malonyl-CoA to yield the phenolic lipids. The phenolic lipids in A. vinelandii were thus found to be synthesized solely from malonyl-CoA by the four members of the ars operon. This is the first demonstration that a type I FAS interacts directly with a type III PKS through substrate transfer.

Abscisic acid controls calcium-dependent egress and development in Toxoplasma gondii

Calcium controls a number of critical events, including motility, secretion, cell invasion and egress by apicomplexan parasites. Compared to animal and plant cells, the molecular mechanisms that govern calcium signalling in parasites are poorly understood. Here we show that the production of the phytohormone abscisic acid (ABA) controls calcium signalling within the apicomplexan parasite Toxoplasma gondii, an opportunistic human pathogen. In plants, ABA controls a number of important events, including environmental stress responses, embryo development and seed dormancy. ABA induces production of the second-messenger cyclic ADP ribose (cADPR), which controls release of intracellular calcium stores in plants. cADPR also controls intracellular calcium release in the protozoan parasite T. gondii; however, previous studies have not revealed the molecular basis of this pathway. We found that addition of exogenous ABA induced formation of cADPR in T. gondii, stimulated calcium-dependent protein secretion, and induced parasite egress from the infected host cell in a density-dependent manner. Production of endogenous ABA within the parasite was confirmed by purification (using high-performance liquid chromatography) and analysis (by gas chromatography-mass spectrometry). Selective disruption of ABA synthesis by the inhibitor fluridone delayed egress and induced development of the slow-growing, dormant cyst stage of the parasite. Thus, ABA-mediated calcium signalling controls the decision between lytic and chronic stage growth, a developmental switch that is central in pathogenesis and transmission. The pathway for ABA production was probably acquired with an algal endosymbiont that was retained as a non-photosynthetic plastid known as the apicoplast. The plant-like nature of this pathway may be exploited therapeutically, as shown by the ability of a specific inhibitor of ABA synthesis to prevent toxoplasmosis in the mouse model.

Influence of temperature and darkness on embryonic diapause termination in dormant Artemia cysts that have never been desiccated

Environmental cues for embryonic diapause termination (EDT) were investigated in the laboratory-produced encysted dormant embryos of the brine shrimp, Artemia franciscana. The cysts were spawned and kept throughout in a 2% sea salt solution. They were activated by various temperatures of the temperate zones or by continuous dark condition (DD), resulting in a state of EDT, and were thereafter able to resume their subsequent development and hatch under appropriate conditions. The level of EDT was conveniently assayed by a hatch test observed within 2 days at 28 degrees C under continuous light condition (LL). A cold treatment of the newly spawned dormant cysts, at 4 degrees C under DD for more than 100 days, resulted in more than 95% hatch of the dormant cysts. Similar treatments of the dormant cysts but at room temperature or 28 degrees C led to significantly different results (30-40% hatch). Almost all the residual non-hatched cysts derived from the above could hatch after an additional cold treatment (at 4 degrees C under DD for about 100 days). This might prove to be latent partial bivoltine in Artemia. Meanwhile, a rearing condition (28 degrees C under LL) induced the newly spawned cysts to hatch scatteredly at and after 1 month, resulting in 22% cumulative hatch on the 92nd day after spawning. When the newly spawned dormant cysts were pretreated at 28 degrees C under DD for 5 or 14 days and then reared at 28 degrees C under LL, the cumulative hatch significantly increased (60%). These results are discussed with respect to probable diapause regulator(s) involved in EDT.

Dinoflagellate diversity and distribution

Dinoflagellates are common to abundant in both marine and freshwater environments. They are particularly diverse in the marine plankton where some cause “red tides” and other harmful blooms. More than 2,000 extant species have been described, only half of which are photosynthetic. They include autotrophs, mixotrophs and grazers. They are biochemically diverse, varying in photosynthetic pigments and toxin production ability. Some are important sources of bioluminescence in the ocean. They can host intracellular symbionts or be endosymbionts themselves. Most of the photosynthetic “zooxanthellae” of invertebrate hosts are mutualistic dinoflagellate symbionts, including all those essential to reef-building corals. Roughly 5% are parasitic on aquatic organisms. The fossil record, consisting of more than 2,500 species, shows a rapid radiation of cysts, starting in the Triassic, peaking in the Cretaceous, and declining throughout the Cenozoic. Marine species with a benthic, dormant cyst stage are confined to the continental shelf and fossil cysts can be used as markers of ancient coastlines. Northern and southern hemispheres contain virtually identical communities within similar latitudes, separated by a belt of circumtropical species. A few endemics are present in tropical and polar waters. Some benthic dinoflagellates are exclusively tropical, including a distinct phycophilic community, some of which are responsible for ciguatera fish poisoning. In lakes chemical and grazing effects can be important. Predatory dinoflagellates co-occur with their prey, often diatoms.

Induction and regulation of host cell-mediated immunity by Toxoplasma gondii.

Toxoplasma gondii is a highly infectious intracellular parasite which, if left unchecked by the immune system, rapidly overwhelms its intermediate hosts, as illustrated by the pathogenesis of toxoplasmic encephalitis in patients with AIDS. In order to insure both its host's and consequently its own survival simultaneously, T. gondii induces a potent gamma-interferon (IFN-gamma)-dependent cell-mediated immunity early in infection that controls the replication of the protozoan and facilitates transformation into the dormant cyst stage. The protective IFN-gamma is derived from three sources: natural killer cells; and CD4+ and CD8+ T lymphocytes, which can partially compensate for each other in knockout mice lacking the appropriate major histocompatibility complex-restricting elements. At least two properties of the parasite appear to be responsible for the early induction of these effector cells. The first is a hydrophobic molecule (or group of related molecules) that triggers interleukin 12 (IL-12), tumour necrosis factor alpha and IL-1beta synthesis in macrophages. This response can also promote HIV replication in the same cells. The second is a superantigen activity that drives IFN-gamma-producing Vbeta5+ CD8+ T cells. These potentially lethal responses are later regulated through the triggering of IL-10 and by the induction of anergy in the superantigen-stimulated Vbeta5+ T cell population.