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The paper deals with methods facilitating the preparation of oncospheres of the cestode, Hymenolepis diminuta, for experimental studies. Described in detail are procedures for the infection of the definitive hosts with the oncospheres; collection and artificial hatching of oncospheres; purification of hexacanths; preparation of extracts from the hexacanths; and preparation of hexacanths for electronmicroscopic studies.
A single pyrimidine nucleoside phosphorylase was found in the cytoplasmic extract from Hymenolepis diminuta. This enzyme preferentially cleaves uridine and, to a much lesser extent, thymidine. Its presence directly indicates the existence of pyrimidine nucleoside salvage pathway in this parasite. Detailed kinetic studies in the phosphorolytic and synthetic direction pointed to the sequential mechanism of these reactions. For phosphorolysis, Kurd = 33 ^M and Kp = 806 nM. For synthesis of uridine, KUra = 204 jiM and Ki.p.rib.= 50 pM. Over six times higher Km for uracil than for uridine indicates that phosphorolysis is the favoured reaction in this tapeworm. Well known inhibitors of mammalian uridine phosphorylase: 2,2'-anhydro-5- -ethyluridine and l-(l,3-dihydroxy-2-propoxymethyl)-5-benzyluracil (DHPBU), both with Ki = 0.07 nM were potent competitive inhibitors of the enzyme from H. diminuta. The newly synthesized 2,3'-anhydro-5-ethyluridine (K. Felczak,unpublished) showed only moderate inhibitory activity (Ki = 14 jiM) similarly as l-(l,3-dihydroxy-2- -propoxy-methyl)-5-benzyluracil. The same order of Ki values obtained for the investigated inhibitors vs uridine phosphorylase, irrespective whether the enzyme was isolated from rat intestinal mucosa (Drabikowska etal., 1987,Biochem. Pharmacol. 36,4125-4128) or H. diminuta may point to a great similarity between binding sites on the parasite and the host enzyme.
Referring to a recently published new taxonomy of the tapeworms, changes in generic combinations, for 19 cestode species recorded in Poland, are proposed.
Acetylcholinesterase (AChE) sequentially extracted from mature specimens of Hymenolepis diminuta was shown to be a globular protein, the monomeric form of which (Ga₁) had molecular mass of 66 kDa as determined by SDS-PAGE. Amphiphilic character of the enzyme was revealed by Triton X-l14 phase partitioning. The cestode AChE preferred acetylthiocholine over propionyl- and butyrylthiocholine as substrate, split N-acetyl-ß-methylthiocholine and myristoylcholine but did not hydrolyze ß-carbonaphthoxycholine, a substrate for butyrylcholinesterases. It was sensitive to 10⁻⁵ M physostigmine and 10⁻⁵ M BW284C51 but not to 10⁻³ M iso-OMPA. No butyrylcholinesterase activity was detected in extracts from the parasite.
Research resulting in the description of six new species representing the genus Fimbriaria has thus revealed that the former Fimbriaria fasciolaris (Pallas, 1781) was in fact a group of species. In the light of this, the long list of final and intermediate hosts for this species may in fact result from earlier errors in identification. Proper diagnosis of Fimbriaria to the level of the species should be based on penetrating morphological studies of adult forms which would take account of: the structure of the oncospheral envelope, the number of genital primordia per segment of the strobila, the number of spines at the cirrus base and the length of the cirrus pouch. Proper diagnosis of species at the larval stage is practically impossible in natural infections because of the great similarity between the larvae of different species. The present state of knowledge of the genus Fimbriaria calls for a revision review of the broad specificity of the whole group in relation to both final and intermediate hosts.
Vitellogenesis in Mosgovoyia ctenoides was examined by means of transmission electron microscopy. Mature vitelline follicles consist of cells in various stages of development, progressing from immature cells of gonial type near the periphery to mature vitellocytes towards the centre. Maturation is characterized by: (1) increase in cell volume; (2) extensive development of large parallel cisternae of granular endoplasmic reticulum (GER), the vitelline material producing units; (3) development of Golgi complexes engaged in vitelline material package; (4) continuous fusion of small vesicles into larger vitelline vesicles and fusion of these into a single very large vesicle, which is characteristic for mature vitellocytes of this tapeworm. Vitellogenesis in M ctenoides is compared with that in other cestodes. Some conclusions concerning the interrelationship between the vitellogenesis pattern and the type of embryogenesis are drawn and discussed.
The envelopes of oncospheres of a hymenolepidid tapeworm with an aquatic life cycle Fimbriaria czaplinskii, surrounding larvae inside the gravid part of the strobila, were examined under a transmission electron microscope. Details of the ultrastructure of the outer envelope and the inner envelope with its two derivatives: the embryophore and oncospheral membrane are described. The ultrastructural features of F. czaplinskii envelopes are compared and discussed with those described previously in the other hymenolepidids, and particularly with the related species Fimbriaria fasciolaris.
The distribution of acetylcholinesterase (AChE) in oncospheres and developing cysticercoids of Hymenolepis diminuta was examined. The enzyme was localized in the nervous system and in some non-nerve cells of these larvae. In oncospheres AChE was detected in hook muscles and in the binucleated medullar center that is known to enclose two neurons. At early developmental stages of the cysticercoids the enzyme was localized in the post-oncospheral hook muscles and in subtegumental muscle fibers of the cercomer. At medium and late stages of development the activity of AChE was detected in the developing nervous system and in two and, subsequently, in four populations of cells, which gradually spread over the whole internal wall of the cyst, thus forming a thin multilayer AChE-positive lining of the cyst cavity. Following withdrawal of the scolex the lining separates the parenchyma of the turned neck from the cyst tissues and remains AChE-positive during the whole life of the parasite, i.e. up to the death of the infected host. The role played by non-neural AChE associated with the cyst cavity lining is unknown, but seems to regulate both the transport of nutrients and minerals into the scolex and waste substances in the opposite direction.
This study includes a redescription of Anoplocephaloides iudicata (Sawada et Papasarathorn, 1966) comb. nov. (Cestoda, Anoplocephalidae) based on museum material from the South-East Asian Tapirus indicus, comparison with Flabelloskrjabinia tapirus (Chin, 1938) from the South American Tapirus terrestris, as well as comments on the genus Flabelloskrjabinia Spasskii, 1951. The present analysis suggests that A. iudicata and F. tapirus are independent species parasitizing T. indicus and T. terrestris, respectively, distinguished primarily by the size and morphology of the scolex and distribution of testes. However, new material from T. terrestris is needed for the evaluation of the generic status of anoplocephalid cestodes from tapirs.
Spermatogenesis in Gallegoides arfaai is similar to that described for other cestode species. Six incomplete synchronic cytokineses occur: four mitotic and two meiotic cell divisions. The primary spermatogonium divides forming two secondary spermatogonia. All further divisions occur simultaneously, resulting in a rosette of four tertiary, then eight quaternary spermatogonia and sixteen primary spermatocytes. The first meiotic division forms thirty-two secondary spermatocytes and after the second meiotic division sixty-four spermatids are formed. Spermiogenesis begins with the formation of a differentiation zone in the form of a conical projection of cytoplasm delimited by a ring of arching membranes. Within this area there are two centrioles, a centriolar adjunct and vestigial striated rootlets. During spermiogenesis, only one of the centrioles develops an axoneme that grows directly into the cytoplasmic extension. The other centriole remains oriented in a cytoplasmic bud and posteriorly aborts. The nucleus elongates and moves into the cytoplasmic extension. Granular material present in each sperm originates from electron-dense material present in the periphery of the spermatid. In the final stage of spermiogenesis two crest-like bodies appear at the Bâse of the spermatid. Finally, the ring of arching membranes constricts and the young spermatozoon detaches from the residual cytoplasm. In order to increase homogeneity in the designation of the non-typical striated rootlets previously described, in this study we propose to group them under the common designation of "vestigial striated rootlets" and its importance is discussed according to previous findings of related structures in other cyclophyllideans.
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