Redescription and molecular characterization of Baruscapillaria spiculata (Nematoda: Capillariidae) parasitizing the Neotropic cormorant Phalacrocorax brasilianus from two Argentinian lagoons.

Two species of intestinal Capillariidae were hitherto known from the Neotropic cormorant Phalacrocorax brasilianus, Baruscapillaria spiculata (Freitas, 1933), and Baruscapillaria appendiculata (Freitas, 1933). The original descriptions are very short and brief, and further reports of both species are scarce and/or confusing. This paper provides a morphological redescription and molecular characterization, based on the partial 18S rDNA gene, of B. spiculata specimens parasitizing the Neotropic cormorant in two continental lagoons from Buenos Aires province, Argentina. Both morphological and morphometrical differences between B. spiculata and B. appendiculata are highlighted on the examination of available type material. Additionally, two previous reports of B. appendiculata from Mexico and Brazil are discussed. A phylogenetic analysis conducted on specimens of B. spiculata and 46 other capillariid isolates available from the GenBank demonstrated a sister-taxon relationship between our specimens and the type species of Baruscapillaria. But, at the same time, significant genetic distances between both taxa showed an interesting variability of the genus Baruscapillaria. The probable division of this genus into multiple genera could probably be confirmed through integrative studies including more species.

Cytological attributes of storage tissues in nematode and eriophyid galls: pectin and hemicellulose functional insights.

Cell walls and protoplast may work together or distinctly in the establishment of the functional profiles of gall tissue compartments. This presumption is herein evaluated in three gall systems by immunocytochemical and ultrastructural analyses. The common storage tissues (CSTs) of leaf galls induced by Eriophyidae on Miconia ibaguensis leaves and by Ditylenchus gallaeformans on M. ibaguensis and M. albicans have rigid and porous cell walls due to their composition of pectins. Hemicelluloses in CST cell walls are scarcer when compared to the cell walls of the control leaves, being functionally compensated by rigid pectate gels. The typical nutritive tissues (TNTs) in galls induced by Ditylenchus gallaeformans are similar to promeristematic and secretory cells regarding their enriched cytoplasm, several mitochondria, and proplastids, as well as multivesicular and prolamellar bodies in cell membranes. The cytological features of the feeding cells of Eriophyidae galls indicate that they are not as metabolically active as the cells of the TNT in nematode galls. However, their cell wall composition suggests more plasticity and porosity than the cells of the TNT, which can compensate the less production of nutrients with more transport. The ultrastructural and immunocytochemical profiles of CST cells reveal functional similarities, which are independent of the taxa of the gall inducer or of the host plant. Despite their analogous functionalities, the protoplast and cell wall features of TNT cells of nematode galls and of the feeding cells of the Eriophyidae galls are distinct, and work out through different strategies toward keeping gall developmental site active.

Imaging dynamic changes of an intracellular cysteine pool that responds to the stimulation of external oxidative stress.

Understanding the dynamic status of intracellular cysteine that responds to the stimulation of external oxidative stress is of great importance to mechanistic studies of cysteine related biology and medicine. However, due to the interference of structural similarity and comparable reactivity of other bio-thiols under physiological conditions, it is exceedingly challenging to develop fluorescent probes with excellent selectivity and sensitivity for the visualization of cysteine activities in an intricate biological context. Herein, we report a new fluorescent probe with an extended coumarin fluorophore, CyP, which displays favorable advantages for intracellular cysteine detection in living cells, and enables effective in vivo visualization of endogenous cysteine activities under external stimuli in live nematodes and plants. In addition, the dynamic changes of the intracellular cysteine pool after exposure to cadmium-induced oxidative stress have been successfully monitored by using this probe in Arabidopsis thaliana, thus providing a valuable diagnostic tool to investigate cysteine functions in a variety of biological phenomena.

A Diffuse Interface Framework for Modeling the Evolution of Multi-cell Aggregates as a Soft Packing Problem Driven by the Growth and Division of Cells.

We present a model for cell growth, division and packing under soft constraints that arise from the deformability of the cells as well as of a membrane that encloses them. Our treatment falls within the framework of diffuse interface methods, under which each cell is represented by a scalar phase field and the zero level set of the phase field represents the cell membrane. One crucial element in the treatment is the definition of a free energy density function that penalizes cell overlap, thus giving rise to a simple model of cell-cell contact. In order to properly represent cell packing and the associated free energy, we include a simplified representation of the anisotropic mechanical response of the underlying cytoskeleton and cell membrane through penalization of the cell shape change. Numerical examples demonstrate the evolution of multi-cell clusters and of the total free energy of the clusters as a consequence of growth, division and packing.

Two independent sulfation processes regulate mouth-form plasticity in the nematode Pristionchus pacificus.

Sulfation of biomolecules, like phosphorylation, is one of the most fundamental and ubiquitous biochemical modifications with important functions during detoxification. This process is reversible, involving two enzyme classes: a sulfotransferase, which adds a sulfo group to a substrate; and a sulfatase that removes the sulfo group. However, unlike phosphorylation, the role of sulfation in organismal development is poorly understood. In this study, we find that two independent sulfation events regulate the development of mouth morphology in the nematode Pristionchus pacificus. This nematode has the ability to form two alternative mouth morphologies depending on environmental cues, an example of phenotypic plasticity. We found that, in addition to a previously described sulfatase, a sulfotransferase is involved in regulating the mouth-form dimorphism in P. pacificus However, it is unlikely that both of these sulfation-associated enzymes act upon the same substrates, as they are expressed in different cell types. Furthermore, animals mutant in genes encoding both enzymes show condition-dependent epistatic interactions. Thus, our study highlights the role of sulfation-associated enzymes in phenotypic plasticity of mouth structures in Pristionchus.

An asymmetric attraction model for the diversity and robustness of cell arrangement in nematodes.

During early embryogenesis in animals, cells are arranged into a species-specific pattern in a robust manner. Diverse cell arrangement patterns are observed, even among close relatives. In the present study, we evaluated the mechanisms by which the diversity and robustness of cell arrangements are achieved in developing embryos. We successfully reproduced various patterns of cell arrangements observed in various nematode species in Caenorhabditis elegans embryos by altering the eggshell shapes. The findings suggest that the observed diversity of cell arrangements can be explained by differences in the eggshell shape. Additionally, we found that the cell arrangement was robust against eggshell deformation. Computational modeling revealed that, in addition to repulsive forces, attractive forces are sufficient to achieve such robustness. The present model is also capable of simulating the effect of changing cell division orientation. Genetic perturbation experiments demonstrated that attractive forces derived from cell adhesion are necessary for the robustness. The proposed model accounts for both diversity and robustness of cell arrangements, and contributes to our understanding of how the diversity and robustness of cell arrangements are achieved in developing embryos.

Hsp90-downregulation influences the heat-shock response, innate immune response and onset of oocyte development in nematodes.

Hsp90 is a molecular chaperone involved in the regulation and maturation of kinases and transcription factors. In Caenorhabditis elegans, it contributes to the development of fertility, maintenance of muscle structure, the regulation of heat-shock response and dauer state. To understand the consequences of Hsp90-depletion, we studied Hsp90 RNAi-treated nematodes by DNA microarrays and mass spectrometry. We find that upon development of phenotypes the levels of chaperones and Hsp90 cofactors are increased, while specific proteins related to the innate immune response are depleted. In microarrays, we further find many differentially expressed genes related to gonad and larval development. These genes form an expression cluster that is regulated independently from the immune response implying separate pathways of Hsp90-involvement. Using fluorescent reporter strains for the differentially expressed immune response genes skr-5, dod-24 and clec-60 we observe that their activity in intestinal tissues is influenced by Hsp90-depletion. Instead, effects on the development are evident in both gonad arms. After Hsp90-depletion, changes can be observed in early embryos and adults containing fluorescence-tagged versions of SEPA-1, CAV-1 or PUD-1, all of which are downregulated after Hsp90-depletion. Our observations identify molecular events for Hsp90-RNAi induced phenotypes during development and immune responses, which may help to separately investigate independent Hsp90-influenced processes that are relevant during the nematode's life and development.

Three-dimensional reconstruction of the pharyngeal gland cells in the predatory nematode Pristionchus pacificus.

Pristionchus pacificus is a model system in evolutionary biology and for comparison to Caenorhabditis elegans. As a necromenic nematode often found in association with scarab beetles, P. pacificus exhibits omnivorous feeding that is characterized by a mouth-form dimorphism, an example of phenotypic plasticity. Eurystomatous animals have a dorsal and a sub-ventral tooth enabling predatory feeding on other nematodes whereas stenostomatous animals have only a dorsal tooth and are microbivorous. Both mouth forms of P. pacificus, like all members of the Diplogastridae family, lack the grinder in the terminal bulb of the pharynx resulting in a fundamentally different organization of several pharynx-associated structures. Here, we describe the three-dimensional reconstruction of the pharyngeal gland cells in P. pacificus based on serial transmission electron microscopical analysis of 2527 sections of 50 nm thickness. In comparison to C. elegans, P. pacificus lacks two gland cells (g2) usually associated with grinder function, whereas the three gland cells of g1 (g1D, g1VL, and g1VR) are very prominent. The largest expansion is seen for g1D, which has an anterior process that opens into the buccal cavity through a canal in the dorsal tooth. We provide the morphological description and fine structural analysis of the P. pacificus gland cells, the behavior of the pharynx and preliminary insight into exocytosis of gland cell vesicles in P. pacificus.

Genome analysis of Diploscapter coronatus: insights into molecular peculiarities of a nematode with parthenogenetic reproduction.

BACKGROUND: Sexual reproduction involving the fusion of egg and sperm is prevailing among eukaryotes. In contrast, the nematode Diploscapter coronatus, a close relative of the model Caenorhabditis elegans, reproduces parthenogenetically. Neither males nor sperm have been observed and some steps of meiosis are apparently skipped in this species. To uncover the genomic changes associated with the evolution of parthenogenesis in this nematode, we carried out a genome analysis. RESULTS: We obtained a 170 Mbp draft genome in only 511 scaffolds with a N50 length of 1 Mbp. Nearly 90% of these scaffolds constitute homologous pairs with a 5.7% heterozygosity on average and inversions and translocations, meaning that the 170 Mbp sequences correspond to the diploid genome. Fluorescent staining shows that the D. coronatus genome consists of two chromosomes (2n = 2). In our genome annotation, we found orthologs of 59% of the C. elegans genes. However, a number of genes were missing or very divergent. These include genes involved in sex determination (e.g. xol-1, tra-2) and meiosis (e.g. the kleisins rec-8 and coh-3/4) giving a possible explanation for the absence of males and the second meiotic division. The high degree of heterozygosity allowed us to analyze the expression level of individual alleles. Most of the homologous pairs show very similar expression levels but others exhibit a 2-5-fold difference. CONCLUSIONS: Our high-quality draft genome of D. coronatus reveals the peculiarities of the genome of parthenogenesis and provides some clues to the genetic basis for parthenogenetic reproduction. This draft genome should be the basis to elucidate fundamental questions related to parthenogenesis such as its origin and mechanisms through comparative analyses with other nematodes. Furthermore, being the closest outgroup to the genus Caenorhabditis, the draft genome will help to disclose many idiosyncrasies of the model C. elegans and its congeners in future studies.

Blood parasites of penguins: a critical review.

Blood parasites are considered some of the most significant pathogens for the conservation of penguins, due to the considerable morbidity and mortality they have been shown to produce in captive and wild populations of these birds. Parasites known to occur in the blood of penguins include haemosporidian protozoans (Plasmodium, Leucocytozoon, Haemoproteus), piroplamid protozoans (Babesia), kinetoplastid protozoans (Trypanosoma), spirochete bacteria (Borrelia) and nematode microfilariae. This review provides a critical and comprehensive assessment of the current knowledge on these parasites, providing an overview of their biology, host and geographic distribution, epidemiology, pathology and implications for public health and conservation.

Tripius gyraloura n. sp. (Aphelenchoidea: Sphaerulariidae) parasitic in the gall midge Lasioptera donacis Coutin (Diptera: Cecidomyiidae).

A new nematode, Tripius gyraloura n. sp., is described from the arundo gall midge, Lasioptera donacis Coutin (Diptera: Cecidomyiidae). This gall midge is being considered as a biological control agent for use in North America against the introduced giant reed Arundo donax (L.) (Poaceae: Cyperales). Thus the present study was initiated to investigate a nematode parasite that was unknown at the time studies with L. donacis were initiated. The new species has a rapid development in the fly host and the mature parasitic female nematodes evert their uterine cells in the hosts' hemolymph. Because large numbers of nematodes sterilise the host, eradication of the parasite from laboratory colonies of the midge may be necessary before populations of the fly are released.

Description of Proyseria petterae n. sp., with an amended generic diagnosis and a review of the species of Proyseria Petter, 1959 and Stegophorus Wehr, 1934 (Nematoda: Acuariidae).

Proyseria decora (Dujardin, 1845) (the type-species of the genus Proyseria Petter, 1959) is redescribed on the basis of specimens from Alcedo atthis (L.) (Coraciiformes: Alcedinidae) from Iran. P. petterae n. sp. is described from Corythornis vintsioides (Eydoux & Gervais) (Alcedinidae) from Madagascar by light and scanning electron microscopy. Proyseria sp. from Alcedo euryzona Temminck from continental Malaysia is described on the basis of a single male specimen. Stegophorus alcedonis Puqin, Yanyin & Guocal, 1991 from A. atthis in China is transferred to the genus Proyseria as P. alcedonis n. comb. The generic diagnosis of Proyseria is amended. Review of the species of the genera Proyseria and Stegophorus Wehr, 1934 is presented.

Bioinformatic analysis of nematode migration-associated genes identifies novel vertebrate neural crest markers.

Neural crest cells are highly motile, yet a limited number of genes governing neural crest migration have been identified by conventional studies. To test the hypothesis that cell migration genes are likely to be conserved over large evolutionary distances and from diverse tissues, we searched for vertebrate homologs of genes important for migration of various cell types in the invertebrate nematode and examined their expression during vertebrate neural crest cell migration. Our systematic analysis utilized a combination of comparative genomic scanning, functional pathway analysis and gene expression profiling to uncover previously unidentified genes expressed by premigratory, emigrating and/or migrating neural crest cells. The results demonstrate that similar gene sets are expressed in migratory cell types across distant animals and different germ layers. Bioinformatics analysis of these factors revealed relationships between these genes within signaling pathways that may be important during neural crest cell migration.

Nematode sperm motility.

Form follows function, and this maxim holds particularly true for the nematode sperm cell. Motility is essential for fertilization, and the process of spermatogenesis culminates in the production of a crawling spermatozoon with an extended pseudopod. However, the morphological similarity to amoeboid cells of other organisms is not conserved at the molecular level. Instead of utilizing the actin cytoskeleton and motor proteins, the pseudopod moves via the regulated assembly and disassembly of filaments composed of the major sperm protein (MSP). The current work reviews the structure and dynamics of MSP filament formation, the critical role of pH in MSP assembly, and the components that regulate this process. The combination of cytological, biochemical, and genetic approaches in this relatively simple system make nematode sperm an attractive model for investigating the mechanics of amoeboid cell motility.

Imaging trematode and nematode parasites.

Recent advances in molecular genetics and in imaging mean that it is now increasingly feasible to image biological processes within helminth parasites and to visualize interactions between worms and their hosts. Moreover, other innovative imaging approaches that are not dependent on transgenic parasites have been applied to, and or developed for, the study of helminth parasites and have provided novel and important insights into the biology of these important pathogens.

Evo-devo of the germline and somatic gonad in nematodes.

Due to recent progress in the development of genetic tools, nematodes have become excellent models to address the mechanistic basis of evolution of development. The gonad is one of the most variable structures in nematodes, reflecting the diverse modes of reproduction and lifestyle in this phylum. During larval development, the gonad primordium has a key role in organizing the neighboring tissues. Therefore, changes in the development of the gonad do not only influence the evolution of its morphology but also the overall body plan of the nematode. Here, we review recent progress on the evolution of development of the germline and somatic gonad in nematodes.

Growth in width and FtsZ ring longitudinal positioning in a gammaproteobacterial symbiont.

Rod-shaped bacteria usually grow in length and place their FtsZ ring and division site at midcell, perpendicular to their long axis [1,2]. Here, we provide morphometric and immunocytochemical evidence that a nematode-associated gammaproteobacterium [3,4] grows in width, sets a constricting FtsZ ring parallel to its long axis, and divides longitudinally by default. Remarkably, the newly described FtsZ ring appears to be not only 90° shifted with respect to model rods, but also elliptical and discontinuous. This reveals an unexpected versatility of the gammaproteobacterial cytokinetic machinery.

Transformation: how do nematode sperm become activated and crawl?

Nematode sperm undergo a drastic physiological change during spermiogenesis (sperm activation). Unlike mammalian flagellated sperm, nematode sperm are amoeboid cells and their motility is driven by the dynamics of a cytoskeleton composed of major sperm protein (MSP) rather than actin found in other crawling cells. This review focuses on sperm from Caenorhabditis elegans and Ascaris suum to address the roles of external and internal factors that trigger sperm activation and power sperm motility. Nematode sperm can be activated in vitro by several factors, including Pronase and ionophores, and in vivo through the TRY-5 and SPE-8 pathways. Moreover, protease and protease inhibitors are crucial regulators of sperm maturation. MSP-based sperm motility involves a coupled process of protrusion and retraction, both of which have been reconstituted in vitro. Sperm motility is mediated by phosphorylation signals, as illustrated by identification of several key components (MPOP, MFPs and MPAK) in Ascaris and the characterization of GSP-3/4 in C. elegans.