The brain structure of selected trypanorhynch tapeworms.

The brain architecture in four species of tapeworms from the order Trypanorhyncha has been studied. In all species, the brain consists of paired anterior and lateral lobes, and an unpaired central lobe. The anterior lobes connect by dorsal and ventral semicircular commissures; the central and lateral lobes connect by a median and an X-shaped crisscross commissure. In the center of the brain, five well-developed compact neuropils are present. The brain occupies a medial position in the scolex pars bothrialis. The ventral excretory vessels are situated outside the lateral lobes of the brain; the dorsal excretory vessels are located inside the brain and dorsal to the median commissure. The brain gives rize four anterior proboscis nerves and four posterior bulbar nerves with myelinated giant axons (GAs). The cell bodies of the GAs are located within the X-commissure and in the bulbar nerves. Highly developed serotonergic neuropils are present in the anterior and lateral lobes; numerous 5-HT neurons are found in the brain lobes including the central unpaired lobe. The X-cross commissure consists of the α-tub-immunoreactive and 5-HT-IR neurites. Eight ultrastructural types of neurons were found in the brain of the three species investigated. In addition, different types of synapses were present in the neuropils. Glial cells ensheath the brain lobes, the neuropils, the GAs, and the bulbar nerves. Glia cell processes form complex branching patterns of thin cytoplasmic sheets sandwiched between adjacent neural processes and filling the space between neurons. Multilayer myelin-like envelopes and a mesaxon-like structure have been found in Trypanorhyncha nervous system. We compared the brain architecture of Trypanorhyncha with that of an early basal cestode taxon, that is, Diphyllobothriidea, and present a hypothesis about the homology of the anterior brain lobes in order Trypanorhyncha; and the lateral lobes and median commissure are homologous brain structures within Eucestoda.

Multiparametric Analysis of Tissue Spheroids Fabricated from Different Types of Cells.

Reproducible, scalable, and cost effective fabrication and versatile characterization of tissue spheroids (TS) is highly demanded by 3D bioprinting and drug discovery. Consistent geometry, defined mechanical properties, optimal viability, appropriate extracellular matrix/cell organization are required for cell aggregates aimed for application in these fields. A straightforward procedure for fabrication and systematic multiparametric characterization of TS with defined properties and uniform predictable geometry employing non-adhesive technology is suggested. Applying immortalized and primary cells, the reproducibility of spheroid generation, the strong correlation of ultimate spheroid diameter, and growth pattern with cell type and initial seeding concentration are demonstrated. Spheroids viability and mechanical properties are governed by cell derivation. In this study, a new decision procedure to apply for any cell type one starts to work with to prepare and typify TS meeting high quality standards in biofabrication and drug discovery is suggested.

Prostaglandins E2 and D2-regulators of host immunity in the model parasite Diphyllobothrium dendriticum: An immunocytochemical and biochemical study.

The spectrum of immunomodulating molecules produced by tapeworms is not yet well understood. The aims of this study, on the tapeworm Diphyllobothrium dendriticum, were: 1) detection and quantification of prostaglandins (PGs) E2 and D2 by high performance liquid chromatography; 2) visualization of PGE2 and PGD2 in specific cells, using methods of immunocytochemistry and confocal laser scanning microscopy; and 3) investigation of the ultrastructure of the cells potentially producing PGE2 and PGD2. The PGE2 immunoreaction (IR) was found in the apical terminals of the frontal glands and sensory organs in the tegument and in small neurons belonging to the main cords and commissures. PGE2-IR partly coincided with α-tubulin-IR. PGD2-IR occurred in the muscle fibers of longitudinal and transverse body muscles and coincided with phalloidin TRITC staining. Both PGE2 and PGD2 were found in the flame cells of the excretory system. Ultrastructural study of the tegument revealed two types of structures that potentially produce PGE2: ciliated and unciliated free nerve endings and frontal gland terminals reinforced with neurotubules. In the main nerve cords, small neurons were identified as potentially exhibiting PGE2-immunoreactivity. In homogenates of the plerocercoids, the measured content of PGE2 and PGD2 was 33.15ngmg-1 and 1.94ngmg-1 of fresh tissue weight, respectively. We found evidence of PGE2 and PGD2 in D. dendriticum parasitizing Coregonus autumnalis (fish) and proved excretion of PGE2 and PGD2 in response to C. autumnalis blood serum. Prostaglandins produced by D. dendriticum probably play a dual role: 1) PGE2 and PGD2 potentially modulate the fish antiparasitic immune response; 2) PGE2 is presumably necessary for proper development and function of the nervous system, and PGD2 can act as an antagonist against mediators causing muscle contraction.

Fine structure of uterus and non-functioning paruterine organ in Orthoskrjabinia junlanae (Cestoda, Cyclophyllidea).

Some cyclophyllidean cestodes provide protection for their eggs in the external environment by providing them with additional protective layers around the egg membranes. In attempting to examine such adaptations, the microanatomy and fine structure of the uterus of pregravid and gravid proglottids of the cyclophyllidean cestode Orthoskrjabinia junlanae, a parasite of mammals that inhabit a terrestrial but moist environment, were studied. In the initial stages of uterine development, developing embryos locate freely in the lumen of a saccate uterus that later partitions into chambers. Each chamber that forms encloses several embryos. The chambers are surrounded by muscle cells that synthesize extracellular matrix actively. The paruterine organs consist of stacks of flattened long outgrowths of muscular cells, interspersed with small lipid droplets. In the gravid proglottids, the size of paruterine organ increases and consists of flattened basal and small rounded apical parts separated by constrictions. The fine structure of the organ wall remains the same: sparse nuclei and stacks of flattened cytoplasmic outgrowths but internal invaginations or lumen in the paruterine organ are absent. Completely developed eggs remain localized in the uterus. Based on the comparative morpho-functional analysis of uterine and paruterine organs and uterine capsules in cestodes, we conclude that these non-functioning paruterine organ in O. junlanae is an example of an atavism. We postulate that the life cycle of the parasite, which infects mammals living in wet habitats, where threats of desiccation of parasite ova is reduced, has favoured a reversion to a more ancestral form of uterine development.

Where are the sensory organs of Nybelinia surmenicola (Trypanorhyncha)? A comparative analysis with Parachristianella sp. and other trypanorhynchean cestodes.

The sensory organs in tegument of two trypanorhynchean species--Nybelinia surmenicola (plerocercoid) and adult Parachristianella sp. (Cestoda, Trypanorhyncha)--were studied with the aim of ultrastructural description and a comparative analysis. The Nybelinia surmenicola plerocercoid lacks papillae with sensory cilia on the bothria adhesive surface. We found an unciliated sensory organ within the median bothria fold. This unciliated free nerve ending contains the central electron-dense disc, three dense supporting rings, and broad root. The nerve ending locates in the basal matrix under the tegument. The tegument of N. surmenicola has a number of ultrastructural features which make it significantly different from other Trypanorhyncha: (i) the tegumental cytoplasm has a plicated constitution in a form of high apical and deep basal folds, (ii) numerous layers of the basal matrix are presented in the subtegument, and (iii) the squamiform and bristlelike microtriches N. surmenicola lack the base and the basal plate. In contrast, numerous ciliated and unciliated receptors were found in Parachristianella sp.: six types on the bothria and one type in the strobila tegument. Ultrastructural constitution of sensory organs in the form of ciliated free nerve endings as well as unciliated basal nerve endings of Parachristianella sp. has many common features inside Eucestoda. In comparison with other Trypanorhyncha, all Nybelinia species studied have less quantity of the bothrial sensory organs. This fact may reflect behavioral patterns of Nybelinia as well as phylogenetic position into Trypanorhyncha. Our observations of living animals conventionally demonstrate the ability of N. surmenicola plerocercoids to locomote in forward direction on the Petri dish surface. The participation of the bothrial microtriches in a parasite movement has been discussed.

Fine structure of the copulatory apparatus of the tapeworm Tetrabothrius erostris (Cestoda: Tetrabothriidea).

The organization and fine structure of the complex copulatory apparatus of Tetrabothrius erostris (Tetrabothriidea) is investigated by light and transmission electron microscopy. A diversity of microstructures was found on the surface of genital ducts. The apical surfaces of male gonadoducts possess tubular and blade-like microtriches that have specific structure in each section of the duct. The apical part of the tubular microtriches contains numerous constrictions in the proximal section of the sperm duct; blade-like microtriches of cirrus possess longitudinal striation in the apical part, and their basal part is reinforced with electron-dense strands. Two types of microtriches occur on the surface of cirrus, and their presence may be considered as systematic features. Prostate glands containing granules of medium electron density (up to 130 nm diameter) are localized in the cirrus sac. The genital atrium contains numerous non-ciliated receptors. Paramyosin-like fibers (up to 200 nm) were found in the muscle fibers surrounding the male atrium canal. Microtriches on the surface of the distal region of the male atrial canal are covered by a glycocalyx. Electron-dense, membrane-like structures (up to 40 nm) lie under the apical membrane of the genital atrium and vagina. These structures do not form a continuous layer; its edges turn down and sink into the apical invaginations of epithelium. Hypotheses on the possible ways of copulation in T. erostris based on the observed ultrastructure are discussed.

Fine structure of the uterus in tapeworm Tetrabothrius erostris (Cestoda: Tetrabothriidea).

The uterine organization in Tetrabothrius erostris (Tetrabothriidea) was investigated by the methods of transmission and scanning electron microscopy. In sexually mature proglottids, the uterine wall consists of a syncytial epithelium (1.4-2.5 µm thick, except in regions containing nuclei). The ribosomes, mitochondria and numerous cisternae of granular endoplasmic reticulum with concentric or parallel profiles with electron lucent material are observed in the epithelium. The uterine wall is characterized by the abundance of lipid droplets that are localized inside the long protrusions of the uterine epithelium (called fungiform papillae) up to 15-17 µm and in the surrounding medullary parenchyma. The protrusions with lipid droplets in the proximal ends of the uterus are located closely to each other. A basal matrix (up to 0.6 µm thick) supports the uterine epithelium. The musculature consisting of 1-2 muscle layers is well developed; large myocytons are connected with the myofibrils and have a nucleus that reaches 4 µm in size. In gravid proglottids, the epithelium without nuclei is reduced to 0.2-1.6 µm thick. The number of protrusions of the uterine epithelium and lipid droplets in the epithelial layer decreases. Sparse small muscle bundles underlay the uterine wall at this stage; the basal matrix is feebly marked. The matrotrophy or the support by nutrition from the parent organism to embryos is discussed for T. erostris which belongs to oligolecital cestodes and possesses numerous lipid droplets in the uterine wall during the development of embryos.

Relationships between uterus and eggs in cestodes from different taxa, as revealed by scanning electron microscopy.

Uterine organization and interaction with developing eggs in Tetrabothrius erostris (Tetrabothriidea), Nippotaenia mogurndae (Nippotaeniidea), Arostrilepis tenuicirrosa, and Monocercus arionis (Cyclophyllidea), cestodes belonging to three different orders, were investigated by scanning electron microscopy. The interactions were traced from sexually mature to gravid proglottids for all species. Pieces of evidence of interactions among these species include specific tight contacts between microlamellae of the uterine epithelium and the egg capsule, networks of fibrils between eggs and uterus, or numerous branched diverticula of the uterine wall that surround eggs or combinations of these. The contacts between uterine epithelium and eggs take place in mature and post-mature proglottids, at a period of development when eggs are newly formed and the embryos are rapidly developing. The eggs grow and develop actively in tight contact with the uterine wall. The maximum diameter of eggs increases 1.5-2 times (or 3.5-4 times in M. arionis) during development. In all species, the intimate contacts between uterus and eggs have weakened or disappeared by the time the proglottids have become gravid. The association between uterus and eggs thus appears as strong evidence of active trophic interaction (or matrotrophy) between the parent organism and developing eggs.

Fine structure of the uteri in two hymenolepidid tapeworm Skrjabinacanthus diplocoronatus and Urocystis prolifer (Cestoda: Cyclophyllidea) parasitic in shrews that display different fecundity of the strobilae.

Adult specimens of Skrjabinacanthus diplocoronatus and Urocystis prolifer were investigated by light and transmission electron microscopy, and data on the uterine organization in these species are presented. The interrelationships of morphological changes of the uterus and developing eggs on the one hand with fecundities of the strobilae of cyclophyllidean cestodes on the other hand are investigated. The development of the excretory system forming a network of excretory ducts close to the uterus is characteristic of S. diplocoronatus. Numerous excretory ducts are located near the uterus or contact the uterine epithelium and are also found in uterine cords. Accumulation of lipids is observed in the cavity of excretory ducts, in muscular cells, and in the lumen of the uterus as well as in the developing eggs. This species displays the largest fecundity among cestode parasites of the shrew. U. prolifer has one of the lowest fecundities among cestodes from shrews. A small amount of lipids found in cells of gravid proglottids could serve as a supply of nutrients. It seems that some cestodes with great fecundity possess specific paths by which eggs are supplied with nutrients. As a result, diverse modifications of the uterus in Cyclophyllidea are formed.