The distribution of Concanavalin A binding sites on the intestinal epithelium of the nematodes Ascans suum and Parascaris equorum

Summary Receptors for Concanavalin A (Con A), were localized on the intestinal epithelium of the nematodes Ascaris suum and Parascaris equorum. Fixed tissue incubated in ³H-Con A showed labeling of the microvilli surface and basal membrane. Using Con A coupled with peroxidase, the tips of the microvilli of Ascaris suum and the tips and lateral surfaces of Parascaris equorum were stained. The basal membrane of both species was also labeled. No labeling was observed on control tissue incubated without Con A or on tissue incubated with Con A to which -methyl-D-mannoside was added.     

Kinetochores and chromatin diminution in early embryos of Parascaris univalens

In Parascaris the mitotic chromosomes of gonial germline cells are holocentric and possess a continuous kinetochore along their entire length. By contrast, in meiotic cells, the centromeric activity is restricted to the heterochromatic tips where direct insertion of spindle microtubules into chromatin without any kinetochore plate is seen. In the presomatic cells of early embryos, which undergo heterochromatin elimination, only euchromatin shows kinetic activity. After developing a technique to separate the very resistant egg shell from the embryos, we studied the cell divisions during early embryogenesis by immunochemical and EM approaches. The results reported here show that in presomatic cells microtubules bind only the euchromatin where a continuous kinetochore plate is present. We also report observations suggesting that the binding of the long kinetochores to the mitotic spindle initiates to a limited number of sites and extends along the entire length, during chromosome condensation. The existence of different centromere stages in different cell types, rends Parascaris chromosomes a very good model to study centromere organization.     

Chromatin diminution in nematode development

Chromatin diminution in Parascaris and Ascaris represents the classical case of a developmentally programmed genome rearrangement. The process is very specific with respect to ontogenetic timing and chromosomal localization, and involves chromosomal breakage, new telomere formation and DNA degradation. Recent evidence from Ascaris lumbricoides var. suum suggests that chromatin diminution might have a function in gene regulation.     

Centromere ultrastructure in germ-line chromosomes of Parascaris

Ultrastructural analysis of the centromere in germ-line mitotic chromosomes of Parascaris univalens and Parascaris equorum revealed that these chromosomes are holocentric. In thin longitudinal sections of both species the kinetochore appeared as a continuous plate (up to 3.8 m long) and displayed a layered structure. This structure consisted of electron-dense inner and outer layers (average width 10 nm) separated by a less dense middle layer (25 nm wide), which had transverse electron-dense bars (10 nm wide) regularly spaced every 25–30 nm. Thus the ladderlike kinetochore profile observed in Parascaris gonial mitotic chromosomes represents a different type of organization from that of the classical trilaminar kinetochore found in both holocentric and monocentric chromosomes.     

Centromere organization in meiotic chromosomes of Parascaris univalens

The chromosomes of Parascaris univalens possess a continuous centromeric structure spanning their entire length in gonial cells. A cytological and ultrastructural analysis of P. univalens meiotic chromosomes was performed. The results show that during meiosis the holocentric germline chromosomes of male P. univalens undergo restriction of kinetic activity to the heterochromatic terminal regions. These regions lack kinetochore structures and interact directly with spindle microtubules.     

Scanning electron microscopy on Parascaris equorum (Goeze, 1782), Yorke and Maplestone, 1926

Research on Ascaridinae from African zebra and zebra living in Vincennes zoological garden (Paris) and those from horses confirmed that they all belong to the same parasite, Parascaris equorum (Goeze, 1782), Yorke &; Maplestone, 1926. Study by scanning electron microscopy permitted greater precision in investigation of various formations: denticules along the peribuccal groove, sinuses separating eulabium from prelabium, intrabuccal sockets.     

Anthelmintic efficacy on <Emphasis Type="Italic">Parascaris equorum</Emphasis> in foals on Swedish studs

Background  

In the last few years stud farms have experienced increasing problems with Parascaris equorum infections in foals despite intensive deworming programs. This has led to the question as to whether the anthelmintic drugs used against this parasite are failing. This study aimed to investigate the efficacy of ivermectin, fenbendazole and pyrantel on the faecal output of ascarid eggs of foals.     

The migration and development of Parascaris equorum in the horse.

Eight worm-free pony foals, aged 2–4 weeks received a single infection of 8000 Parascaris equorum eggs. The foals were killed at intervals to investigate the migration and subsequent development of P. equorum. Signs of larval invasion were found in the liver within 48 h after infection. Larvae migrated from the liver to the lungs from 7 to 14 days after infection and the majority had returned to the small intestine via the tracheooesophageal route by day 23.After completion of their tissue migration, the parasites grew rapidly in size in the lumen of the small intestine, but after day 37 the total number of worms present in this site decreased. Patent infections developed on days 101–104.     

Toxocara infection in pigs. The use of indirect fluorescent antibody tests and an in vitro larval precipitate test for detecting specific antibodies.

An in vitro larval precipitate test using second-stage Toxocara canis larvae and an indirect fluorescent antibody (IFA) test employing cuticles of T. canis larvae as antigen were evaluated using antisera produced in pigs experimentally infected with T. canis, T. cati, Ascaris suum, Toxascaris leonina and Parascaris equorum. The former test was both specific and sensitive and is suggested as a reliable and simple method of detecting Toxocara antibodies in pigs. The latter test was considered unsuitable because of cross-reactions that occurred when sera from pigs infected with other ascarids were tested. An IFA test for Ascaris antibodies, employing cuticles of A. suum larvae as antigen, is described. The degree of specificity of this test suggests that it may be of value in the detection of antibodies to Ascaris in pigs under natural conditions.     

The sperm-egg fusion in the nematode Parascaris equorum

Summary

The process of fertilization and the sperm storage in the female apparatus in Parascaris equorum is described in this paper. The sperm approaches the egg by means of pseudopodia containing bundles of microfilaments. The sperm and egg membranes fuse and the sperm penetrates progressively into the ovum. The egg and sperm plasma membranes and glycocalyces disappear at the point of fusion. At the end of fertilization, they are reformed at the egg's surface, while the egg and sperm chromatin begins to decondense. Spermatozoa are stored in the female apparatus prior to fertilization; here they come into contact with the epithelial cells of the spermatheca, protruding pseudopodia rich in microfilaments into the cellular body.     

Chromatin diminution in the parasitic nematodes ascaris suum and parascaris univalens

Chromatin diminution in Parascaris univalens and Ascaris suum undoubtedly represents an interesting case of developmentally programmed DNA rearrangement in higher eukaryotes. It is a complex mechanism involving chromosomal breakage, new telomere addition and DNA degradation, and occurs in all presomatic cells. The process is rather specific with respect to its developmental timing and the chromosomal regions that are eliminated. The functional significance of chromatin diminution still remains an enigma. The fact, however, that single-copy, protein-coding genes are contained in the eliminated DNA demonstrates that in P. univalens and A. suum, there is a qualitative difference between germ-line and somatic genomes, and suggests that chromatin diminution may be used as a "throw-away" approach to gene regulation. We present a hypothesis as to how, during evolution, a partial genome duplication might have been linked to the process of chromatin diminution, in order to provide a selective advantage to parasitic DNA-eliminating nematodes.     

Protease inhibitors from the parasitic worm Parascaris equorum

Two proteic inhibitors (I and II) of serine proteases have been purified from the parasitic worm Parascaris equorum by affinity chromatography on immobilized trypsin followed by preparative electrophoresis. They have an apparent relative molecular mass of 9000 and 7000 as determined by gel filtration, a slightly acid isoelectric point (5.5 and 6.1) and a similar amino acid composition. Both inhibitors lack serine, methionine and tyrosine. They bind bovine trypsin extremely strongly with an association constant, Ka, larger than 10(9) M-1, and form a 1:1 complex with this protease. The Ka values for the binding to bovine chymotrypsin are approximately 3.3 X 10(8) M-1 (inhibitor I) and approximately 2 X 10(6) M-1 (inhibitor II). Inhibitor I interacts also with porcine elastase (Ka approximately 5 X 10(7) M-1), while inhibitor II is inactive towards this enzyme.     

A survey in Louisiana of intestinal helminths of ponies with little exposure to anthelmintics

Ponies reared with minimal or no exposure to anthelmintics were surveyed for intestinal helminths in order to estimate prevalence and intensity of parasite populations unaltered by frequent exposure to anthelmintics. Thirty-seven mixed breed ponies of varying ages were examined. Thirty-four species of nematodes and 2 species of cestodes were found. Twenty-four of the nematode species (including 1 new species) were in the subfamily Cyathostominae (small strongyles). Eighty-seven percent of the total burden of adult small strongyles in the large intestine was composed of 10 species. By comparing the results of the present survey with those of recent surveys of horses from herds which had been subjected to treatments with anthelmintics, the effect of prolonged usage of anthelmintic treatment on the prevalence of individual species possibly can be estimated. The general ranking of the 10 most common cyathostome species was similar to those described in recent surveys of horses, suggesting that anthelmintic pressure does not affect the prevalence of most cyathostome species. The lack of anthelmintic treatment appeared not to affect prevalence rates for Anoplocephala perfoliata and Anoplocephala magna when compared to other studies. Conversely, prevalence rates for Strongylus spp., Triodontophorus spp., Craterostomum acuticaudatum, Oxyuris equi, and Parascaris equorum were higher than those reported for these species in recent studies of horses.     

Biochemical and immunological systematics of some ascaridoid nematodes: genetic divergence between congeners

Vertical starch gel electrophoresis and trefoil immunodiffusion were used to study the systematics of some ascaridoid nematodes. Within the Ascarididae, the time scale of divergence was too great for intergeneric electrophoretic comparisons. Congeneric electrophoretic comparisons of Baylisascaris procyonis (host--raccoon) versus Baylisascaris transfuga (host--black bear), and Toxocara canis (host--domestic dog) versus Toxocara cati (host--domestic cat) yielded Nei genetic distance coefficients of 1.21 and 1.55, respectively. Estimates of times of divergence made from 1 electrophoretic clock calibration suggest that the Baylisascaris species have not shared a common ancestor for 25 million years (Myr), and that the Toxocara species diverged 33 Myr ago. The Baylisascaris divergence estimate corresponds to host-family divergence estimates based on immunological and paleontological evidence, which suggests that cospeciation has occurred. In contrast to this, Ascaris suum (host--pig) and Ascaris lumbricoides (host--human) have a distance coefficient of 0.09. This indicates that these species diverged comparatively recently and may represent a case of host range expansion. Trefoil immunodiffusion comparisons of ascaridoid albumins yielded reactions of identity for A. suum, A. lumbricoides, Parascaris equorum, B. procyonis, B. transfuga, T. canis, and T. cati. This confirms that these taxa are members of a monophyletic group.     

Seasonal transmission of gastrointestinal parasites of equids in southern Louisiana.

Parasite-naive pony foals were used as sentinels to monitor transmission of gastrointestinal parasites of equids in Louisiana during 4 seasons of the year. Two annual periods were studied, 1988-1989 and 1992-1993. Two or 3 foals each season were turned out to graze a contaminated pasture along with resident parasitized mares and their foals. After a grazing period of 8 wk, sentinel ponies were held in a parasite-free box stall for a period of 6 wk to allow parasites to develop, thus enhancing the evaluation of hypobiotic stages. Following this holding period, necropsies were performed for complete parasite recoveries. Data show that transmission of large and small strongyles occurs during all seasons in southern Louisiana, with highest levels of transmission occurring in the winter and only minimal transmission taking place in the summer. Numbers of mucosal cyathostomes, as well as total cyathostome numbers, were highest in the winter, and luminal cyathostome numbers were highest in the spring. Transmission of Anoplocephala perfoliata and Parascaris equorum occurred during all seasons of the year, although numbers of P. equorum were reduced in spring 1989 and 1993. Gasterophilus intestinalis instars were recovered from fall and winter sentinels only. Oxyuris equi L4 were found all seasons 1 yr, but only during the fall and winter of the final year.     

Cytological analysis of chromosomes in the two species Parascaris univalens and P. equorum

Chromosome organization and the phenomenon of chromatin diminution in the two species Parascaris univalens (2n=2) and P. equorum (2n=4) were cytologically analysed by a variety of staining techniques (Quinacrine, Hoechst 33258, Chromomycin A₃ and C-banding). The results show that: (1) the chromosomes of the two species differ markedly in both the location and the type of heterochromatin they contain; (2) in both species there is a strong chromosome polymorphism which, however, ranges within a basic species-specific phenotype; (3) the heterochromatin can be eliminated in presomatic cells during early embryogenesis at two different stages and in both cases the consequence of this process is the generation of somatic cells with a 2n=60 karyotype. Moreover, evidence suggesting the sterility of hybrids between the two species is provided.     

Transglutaminase activity in equine strongyles and its potential role in growth and development.

Transglutaminases (E.C. 2.3.3.13) are a family of Ca(2+)-dependent enzymes that stabilize protein structure by catalyzing the formation of isopeptide bonds. A novel form of transglutaminase has been identified and characterized that seem to play an important role in growth, development, and molting in adult and larval stages of filarial nematodes. The aim of this study was to identify the ubiquitous nature of this enzyme in other nematodes and to measure its significance to larval growth, molting, and development. For this purpose, equine Strongylus spp. were used. Activity of this enzyme was identified in extracts of larvae and adults of Strongylus vulgaris, S. edentatus, Parascaris equorum and Cylicocyclus insigne. The significance of transglutaminase in the early growth and development of Strongylus vulgaris, S. edentatus and S. equinus was tested by adding specific inhibitors, monodansylcadaverine (MDC) or cystamine (CS), to in vitro cultures of third (L3) and fourth stage larvae (L4). The viability, molting and growth of these nematode species were affected by both inhibitors. Cystamine promoted abnormal development of Strongylus edentatus L3, resulting in an aberrant expansion of the anterior end. Addition of these inhibitors to cultures of L4 also reduced growth of the three species. The results indicated that transglutaminase is present in a wide array of nematode parasites and may be important in growth and development of their larval stages.     

Gastrointestinal helminths of ponies in Louisiana: a comparison of species currently prevalent with those present 20 years ago

A survey in Louisiana of gastrointestinal helminths recovered at necropsy from 117 ponies with minimal exposure to anthelmintics between 1989 and 2000 is compared with a survey conducted 20 yr earlier in the same region. An objective of this study was to determine whether species diversity has been affected by the advent and use of the macrocyclic lactone (ML) parasiticides and by the increased anthelmintic pressure on the helminth species infecting the general equine population. Twenty-six cyathostome species and 8 strongyle species were recovered. Two cyathostome species that were not found before, Cylicostephanus asymetricus and C. bidentatus, and 1 species of large strongyle, Oesophagodontus robustus, were added to the list of species found in Louisiana. All cyathostome and large strongyle species found previously were still present. But prevalences and intensities were significantly reduced for almost all large and small strongyle species. Prevalences and intensities of Oxyuris equi adults and larvae were reduced, whereas the prevalence of Parascaris equorum remained constant. The tapeworm Paranoplocephala mamillana was added to the list of parasite species found in Louisiana. Anoplocephala perfoliata remained the most common cestode. This species was found at the same level of intensity but increased slightly in prevalence. Anoplocephala magna was found less frequently than previously. The overall diversity of species remained reatively unchanged. The reasons for the differences in intensity and prevalence of strongyles between these 2 periods are unknown but might be related to the development and use of the broad-spectrum ML anthelmintics in the intervening period, a difference in the population of equids surveyed, different techniques used to identify the parasites, or differences in numbers of parasites identified (or to all).