Exposed and hidden lectin-binding epitopes at the surface of Borrelia burgdorferi.

The presence of surface- and subsurface-located lectin-binding epitopes of Borrelia burgdorferi was examined by electron microscopy using a variety of gold-labeled lectins. Concanavalin A reacted predominantly with extracellular material adjacent to the spirochetes. Wheat germ agglutinin bound weakly to the surface of borreliae; however, alterations of the outer membrane by preincubation in 100 ppm Triton X-100 or boiling uncovered numerous periplasmic sites recognized by the lectin. The periplasmic flagella liberated by some cells after detergent treatment were labeled with concanavalin A, wheat germ agglutinin and Ulex europaeus agglutinin UEA-I. No surface-exposed or periplasmic epitopes for the lectins from Glycine max, Dolichos biflorus or Helix pomatia were detected.

Presence and cellular distribution of soybean agglutinin-binding epitopes in two strains of Bacillus subtilis.

The cellular location of N-acetylgalactosamine in Bacillus subtilis strains 168 and 170 was examined by electron microscopy using gold-conjugated soybean agglutinin (SBA) as a marker. Post-embedding labeling of sectioned material showed SBA-reactive, galactosamine-containing polymers associated with the cell membrane or the cytoplasm of the two strains. This intracellular location was distinct from the concanavalin A-binding epitopes that were located over the cell wall. Labeling of whole cells (native, fixed in glutaraldehyde, or treated with proteinase K, or Tween 20) before negative staining revealed no galactosamine exposed on the surface of strain 168. On the surface of strain 168 some exposed galactosamine terminal residues were detected; their accessibility to SBA increased when Tween 20 or proteinase K was applied.

Ultrastructure of spermiogenesis and the mature spermatozoon of Tetrabothrius erostris Loennberg, 1896 (Cestoda, Tetrabothriidae).

The spermiogenesis of Tetrabothrius erostris is characterized by the following events: formation of a differentiation zone containing 2 basal bodies and a pair of rootlets; one of the basal bodies gives rise to a free flagellum, the other induces formation of a flagellar bud; rotation at 90 degrees of the flagellum prior to its fusion with the middle cytoplasmic process of the differentiation zone and partial rotation of the flagellar bud; penetration of the nucleus between the rootlets and appearance of a spur-like protrusion in the differentiation zone; elongation and twisting of the differentiation zone, resulting in twisting of the peripheral microtubules and migration of the nucleus; formation of a crested body; proximal densification of the spermatozoon prior to its detachment from the spermatid rosette. The mature spermatozoon has a single axoneme of 9+"1" type and twisted peripheral microtubules. It consists of 3 portions: a proximal part with a crested body, a middle region rich in beta-glycogen, and a distal part containing the nucleus. The pattern of spermiogenesis resembles most closely that in phyllobothriid tetraphyllideans, and probably reflects a relationship of the family Tetrabothriidae with this group.

Grillotia erinaceus (Cestoda, Trypanorhyncha): localization of neuroactive substances in the plerocercoid, using confocal and electron-microscopic immunocytochemistry.

Indirect immunocytochemistry, in conjunction with confocal scanning laser microscopy and electron-microscopic immunogold labeling, has been used to localize neuropeptide and 5-hydroxytryptamine (5-HT) immunoreactivities (IRs) in the plerocercoid (scolex and surrounding blastocyst) of the trypanorhynch tapeworm, Grillotia erinaceus. Antisera directed to two native cestode neuropeptides, neuropeptide F and the FMRFamide-related peptide, GNFFRFamide, were used to demonstrate the presence of a well-developed and extensive peptide-immunoreactive nervous system of central and peripheral elements in the juvenile scolex. Neuronal connectivity exists between the scolex and the surrounding blastocyst, in which there is a rich innervation of varicose fibers displaying peptide IR. Ultrastructurally, gold labeling of the peptide IR was found exclusively over the contents of dense secretory vesicles in the axons and somatic cytoplasm of neurons. Double-labeling experiments demonstrated an apparent colocalization of peptide IR, although the results of antigen preadsorption procedures indicated substantial cross-reactivity of the two antisera. A separate and well-differentiated 5-HT-immunoreactive nervous system, with a similar anatomical arrangement as the peptide-immunoreactive nervous system, is present in both the scolex and blastocyst.

Effects of luxabendazole on the tegument of Fasciola hepatica.

The effects in vivo of 5, 10, and 20 mg/kg of luxabendazole (LBZ) on the tegument of Fasciola hepatica have been examined 48 h, 7 days and 14 days post-treatment of experimentally-infected rats. As early as 48 h post-treatment, the drug is shown to provoke significant damage to the tegument. The pathological phenomena characterizing LBZ damage are blebbing of the apical plasmalemma, formation of microvillus-like projections over the free surface, swelling of the basal infolds and stimulation of autophagy. The spines are often fractured; the tegument in the vicinity of spines seems more strongly altered than that in other foci. The basal layer is often changed, from increase of electron density to lack of integrity with the apical cytoplasm. The progress of the ultrastructural damage with time is not expressed. However, cytochemical data show that a longer post-treatment intervals the surface-coat structure becomes irregular and patches of ruthenium red positive material of variable thickness are focally accumulated. Only a slight dose-effect is noted 48 h after LBZ application when the alterations provoked by 5 mg/kg are less evident than those by 10 and 20 mg/kg.

Effects of three anthelmintics on the tegument of Hymenolepis fraterna (Cestoda).

The in vivo effects of the anthelmintics taenifugin, VUFB 14170 and VUFB 15269 on the tegument of Hymenolepis fraterna have been examined by SEM, TEM and cytochemistry. The drugs were given to H. fraterna-infected mice on the 14th day post-infection in a single oral dose of 150, 200 and 200 mg/kg, respectively. By 72 h post-treatment, the drug-induced pathomorphological changes in the tegument indicated that all three drugs had a significant effect. Changes were most pronounced on the brush border and in the intercellular material. On the apical surface, there was blebbing as well as accumulation of membrane fragments over the microthrix tips and erosion of the brush border. The intercellular material was changed in structure, showing increased electron density in some areas and oedema of the intercellular spaces in other areas. There were also fractures of the tegument of variable depth, sometimes reaching to the parenchyma. These results suggest altered tegumental integrity and, occasionally, complete disruption of the selective permeability barrier created by the normal tegument. This suggestion is further supported by the penetration of ruthenium red into some tegumental areas and its distribution into the intercellular spaces, down to the parenchyma. The intrategumental lysosomes also appeared to be significantly activated. There was evidence of autophagy in both distal cytoplasm and tegumental cells. Mature and gravid proglottides were more susceptible to drug damage than those in the anterior strobila and neck.