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By use of a thin-layer chromatogram (TLC) overlay procedure, 35S-labeled acanthamoebae were shown to bind to seven glycolipids of rabbit corneal epithelium. Corneal epithelial cells were grown in culture and were subjected to Folch extraction to isolate a chloroform-rich lower phase containing neutral glycosphingolipids (NGSL) and an aqueous upper phase containing gangliosides, i.e., sialic acid-containing glycolipids. Thin-layer chromatography of the upper phase revealed the presence of 10 ganglioside components. Acanthamoebae were shown to bind to four of these components, referred to as 2, 3, 6, and 7. On TLC plates, ganglioside components 2 and 3 migrated slightly ahead of the glycolipid standard GD1a, component 7 comigrated with standard GM3, and component 6 migrated a little more slowly than GM3. Likewise, of the 10 NGSL known to be present in the lower phase, acanthamoebae bound to components 1, 5, and 6. NGSL components 1, 5, and 6 migrated on TLC plates with relative mobilities similar to those of standards asialo GM1, asialo GM2, and ceramidetrihexoside, respectively. We propose that one or more of the Acanthamoeba-reactive glycolipids of corneal epithelium identified in this study may play a role in the pathogenesis of Acanthamoeba keratitis by mediating the adherence of the parasites to the cornea.

Scanning and electron microscopy was used to study the pathogenesis that occurred in mouse epithelial cells that had been penetrated by Eimeria papillata sporozoites. Optimal penetration of parasites injected into nonligated and ligated mouse intestine was found to occur at 4-15 min post-inoculation. During initial penetration, the parasite caused disruption of the microvilli of the intestinal cells, which led to detachment of the microvilli from the plasma membrane of the penetrated cell. Host cells penetrated by the parasite showed extensive destruction of the internal cellular organization together with blebbing of host-cell cytoplasm and release of internal organelles such as mitochondria. Ultimately, the penetrated cells completely broke down, leaving vacuolated areas next to ultrastructurally normal epithelial cells.

1. To further characterize the basic aspects of parasite-cell interaction, untreated and hormone-treated Madin-Darby canine kidney (MDCK) cells were allowed to interact with T. vaginalis. 2. Among the six hormones assayed, alpha-estradiol caused MDCK cells to become highly adhesive to T. vaginalis. Increases in parasite adhesion of 83% and 68% were obtained by previous incubation of the epithelial monolayers for 18 h with 1 ng/ml alpha-estradiol and 10 ng/ml 17-beta-estradiol, respectively. 3. The estrogenic response of MDCK cells was inhibited by tamoxifen, cycloheximide, puromycin, or 2-deoxy-D-glucose. 4. We conclude that MDCK cells possess receptors for estrogens and that T. vaginalis cytoadherence can be influenced by the hormonal cycle of its host.

Toxoplasma gondii-like schizonts were found in tracheal epithelium of an 8-yr-old male cat. The parasites were located in parasitophorous vacuoles within the host cell cytoplasm, divided by schizogony, contained periodic acid-Schiff-positive granules, and reacted with anti-T. gondii serum but not with anti-Neospora caninum serum. Mature schizonts were 7.0 x 5.9 microns (5-10 x 4-10 microns; n = 22) and contained 4-16 merozoites. The merozoites were approximately 5 x 1 microns.

Interactions of pathogenic Entamoeba histolytica (HM 1) with human intestinal epithelial cells (Henle-407) were investigated. The E. histolytica trophozoites adhered and cytolysed 87% of cultured epithelial cell monolayers. A significant (P less than 0.001) inhibition of cytopathic effect of amoebic trophozoites pretreated with monoclonal antibodies to a 29 kDa surface associated protein suggested utilization of the 29 kDa surface protein in recognition and cytolysis of epithelial target cells. The polyclonal sera from treated patients of amoebic liver abscess and anti-amoebic hyperimmune serum inhibited cytopathogenicity to a greater degree (P less than 0.001) than did the monoclonal antibodies. The data thus suggest involvement of several amoebic molecules in exercising cytopathogenicity to epithelial cells.

Human corneal buttons were exposed to Acanthamoeba castellanii trophozoites and cysts for 12 hours at 35 degrees C. The buttons examined by light microscopy and scanning and transmission electron microscopy had severe epithelial ulceration and penetration by trophozoites. Observations on trophozoites below the surface suggest that penetration is accomplished by both secreted cytolytic enzymes and phagocytosis. It is likely that the secretion of one or more enzymes constitutes the initial step in preparing the host tissue for endocytosis or that the secretory mechanism is used by the amebas to move through the outer squamous layer to the basement epithelium where phagocytosis occurs. Based on this study and a previous study, it appears that entry into the cornea is a two-step process involving adherence and penetration by trophozoites.

Human corneal buttons were exposed to trophozoites and cysts of Acanthamoeba castellanii for 12 hours. Examination of the buttons by scanning electron microscopy showed numerous trophozoites on the surface of the epithelium. Trophozoites examined by transmission electron microscopy had limited regions of attachment to the epithelium but extensive regions of attachment to each other. Attachment regions were characterized as plaque-like maculae of an incomplete desmosome junction. Firm attachment mechanisms may explain how penetration of the human cornea occurs.

When sporozoites of Eimeria tenella were inoculated into an isolated horn of the bicornuate caecum of the domestic fowl, replicating infections, culminating in the production of oocysts, were found, almost exclusively, in the inoculated horn. In contrast, sporozoites of E. tenella introduced into the bloodstream via a superficial vein induced infections of equivalent intensity in both horns of the caecum. These findings are discussed with reference to (1) the detection (by tissue transfer) of infective sporozoites in samples of cardiac blood and vascular tissues taken within 3 to 18 h of the oral inoculation of oocysts, and (2) the (previously published) observation that the sporozoites of E. tenella are transported from the superficial epithelium of the caecum (site of invasion) to the crypts (site of initial development) within host intra-epithelial lymphocytes. We suggest that a local inflammatory response to the invasion of sporozoites may be responsible for their re-location in proximity to the site of entry. This could act by directing the homing (or retention) of the parasite-bearing lymphocytes, and/or by providing a stimulus for the sporozoite to leave its transporting cell.

Microsporidia are obligate intracellular protozoan parasites that are becoming increasingly recognized as opportunistic pathogens in patients with AIDS. They have been associated with enteritis, hepatitis, and peritonitis and recently keratoconjunctivitis. Gram stain demonstrates the presence of these organisms on light microscopic sections. The specific diagnostic features that distinguish microsporidia from other small nonspore-forming organisms are best demonstrated by electron microscopy, which is also used to characterize the members of Microsporea. In this study, salient histopathologic features of microsporidia in corneal epithelium obtained from an HIV-seropositive individual who developed AIDS are illustrated and discussed.

The growth of Eimeria vermiformis within cultured murine fibroblastlike (L-929) or rat epithelial-like (RATEC) cells was inhibited by treatment of the cells with the appropriate recombinant gamma interferon. The effect was apparent as a reduction in both the initial numbers of intracellular sporozoites and, to a much greater extent, the numbers of subsequent developmental stages. Pretreatment of the host cells was more effective than treatment in the early postinvasive period, and recombinant gamma interferon had no effect on the development of the parasite if added 24 h or later after the inoculation of sporozoites. Incubation of sporozoites in medium containing recombinant gamma interferon in no way affected their ability to invade or to grow within host cells. These findings indicate that the inhibitory effects of recombinant gamma interferon on the growth of E. vermiformis are mediated via the host cell and are directed mainly against the transforming sporozoite, although the ability of the sporozoite to invade the host cell was also reduced to some extent. The later developmental stages were refractory to the effects of this lymphokine.

We have used glutaraldehyde-fixed target cells to study the attachment phase of cell invasion by live trypomastigotes of Trypanosoma cruzi, and determined that attachment is polarized and receptor-mediated. T. cruzi trypomastigotes bind much less efficiently to confluent epithelial cells, which are polarized, than to sparse epithelial cells. When the tight junctions of confluent epithelial cells are disrupted by removing Ca2+ from the incubation medium before glutaraldehyde fixation, binding of T. cruzi increases. T. cruzi also shows preference for attachment underneath cells or to the edges of cells. The binding occurs within a few minutes, is saturable, and is influenced by the parasite developmental stage. Fab fragment derived from monoclonal antibodies that immunoprecipitate a 160-kDa molecule present only on the surface of trypomastigotes inhibit adhesion to fixed and live cells. Future characterization of the target cell receptors for this molecule and the use of fixed target cells should facilitate studies of the mechanisms involved in the initial interaction of T. cruzi with its host cells.

The ultrastructural development of Eimeria tenella was studied in experimentally infected chicks fed 90 ppm lasalocid, an ionophorous anticoccidial antibiotic. Drug treatment was timed to target-specific endogenous stages. At 6 h after infection, many sporozoites within the epithelium showed degradation as a result of drug action. Only a few intact sporozoites were seen. The drug caused outer-membrane blistering, large surface swellings and enlarged mitochondria in both first- and second-generation merozoites. No effect on the gamonts was discerned.

Various conditions of cultures were performed to investigate the role of tight junctions formed between adjacent MDCK cells on the entry of Toxoplasma. When MDCK cells were cocultured with excess number of Toxoplasma at the seeding density of 1 x 10(5), 3 x 10(5), and 5 x 10(5) cells/ml for 4 days, the number of intracellular parasites decreased rapidly as the host cells reached saturation density, i.e., the formation of tight junctions. When the concentration of calcium in the media (1.8 mM in general) was shifted to 5 microM that resulted in the elimination of tight junction, the penetration of Toxoplasma increased about 2-fold (p less than 0.05) in the saturated culture, while that of non-saturated culture decreased by half. Trypsin-EDTA which was treated to conquer the tight junctions of saturated culture favored the entry of Toxoplasma about 2.5-fold (p less than 0.05) compared to the non-treated, while that of non-saturated culture decreased to about one fifth. It was suggested that the tight junctions of epithelial cells play a role as a barrier for the entry of Toxoplasma and Toxoplasma penetrate into host cells through membrane structure-specific, i.e., certain kind of receptors present on the basolateral rather than apical surface of MDCK cells.

This study compared the tissue distribution and cellular expression of Ia antigen in jejunal and ileal epithelium at various stages of intestinal inflammation produced by infecting rats with the nematode parasite Nippostrongylus brasiliensis. Tissues were examined at Day 0 (control), Day 4 (early), Day 10 (acute), and Day 16 (recovering). Frozen sections were stained with the MRC OX-4 anti-Ia monoclonal antibody using an immunoperoxidase technique. Control jejunal sections demonstrated positive epithelial Ia expression mainly in the mid-regions of the villi. The stain appeared to be mostly intracellular in the supranuclear area; the basolateral membrane stained faintly. At Day 4, a greater percentage of the epithelial cells expressed Ia, including those at the tips of the villi. The Day 10 sections demonstrated no staining at all of villus enterocytes, but the crypt epithelium was Ia positive. At Day 16, the pattern of Ia expression was similar to that seen in the early infection. In the ileum, stain was present in enterocytes over most of the villus and crypt regions except in the villus-crypt junction and did not change significantly during infection. We conclude that the changes in the expression of Ia antigen by intestinal epithelium are local to the site of infection and probably occur as a consequence of the host's inflammatory response.

Adherence of trichomonads to host epithelial cells appears to be a critical step in the pathogenesis of trichomoniasis. We evaluated the effect of a panel of 10 monoclonal antibodies on attachment of [35S]methionine-radiolabeled Trichomonas vaginalis strains to HeLa cell monolayers. Of 10 monoclonal antibodies, 3 totally eliminated motility of PHS2J strain trichomonads and reduced their adherence to 48 to 60% of control values (P less than 0.001). However, none of the monoclonal antibodies affected motility or adherence of STD13 strain trichomonads. Although the antibodies all reacted with PHS2J trichomonads by immunofluorescence, there was no correlation between inhibition of adherence and findings on either immunofluorescence or radioimmunoprecipitation. Direct microscopic observations showed that incubation with the monoclonal antibodies did not cause cytolysis of T. vaginalis. In quantitative cultures there was no difference in the number of colonies produced by parasites that had been incubated with antibodies that inhibited or had no effect on adherence. We conclude that our monoclonal antibodies reduced adherence not by cytotoxic effects or by competing for specific sites mediating adherence of the protozoa, but by inhibiting motility of T. vaginalis.

The behaviour and pathogenic effects of Trichomonas vaginalis in mammalian cell cultures were studied using light microscopy and scanning electron microscopy. Six hours after inoculation of the parasites into the cell cultures about 10% of the epithelial monolayer was destroyed. The parasites adhered to the epithelial cells, developed an amoeboid morphology, and crawled over and under the monolayer of cells. These observations suggest that the adhesiveness, amoeboid morphology, and motility of T vaginalis may be important mechanisms in the injury caused to the vaginal epithelium.

Giardiasis is the most common waterborne diarrheal disease in the United States and is highly prevalent throughout the world. The clinical spectrum of disease ranges from asymptomatic infection to persistent severe malabsorption. The precise interaction between Giardia and its human host remains conjectural because of the paucity of published studies that address the details of its pathogenesis. The immune system of the host responds to this protozoan parasite, and the intestinal epithelium is a site of interaction between parasite and host. Possible mechanisms whereby Giardia may alter the host's absorption of nutrients at the epithelial level include direct physical interference, toxin secretion, direct physical alteration of the epithelium, competition for nutrients, induction of an inflammatory response, and coincidental infection of the host with a second organism. The host's immune system may play both a protective and a pathogenic role.

A new species, Demodex foveolator, is described in all stages of the life cycle from the lesser white-toothed shrew, Crocidura suaveolens (Pallas, 1821). The species inhabits epidermal pits and apparently is a low-grade pathogen. A suggested trait of structural adaptations of epithelia-dwelling demodicids to their habitats is briefly discussed.

Germ-free guinea pigs were inoculated intracecally with Entamoeba histolytica and the enteric flora derived from a human patient with acute amebic colitis. Aanimals were killed at post-inoculation intervals of 7 to 12 days. The mode of penetration of cecal epithelium by the ameba was examined by light and electron microscopy. The following sequence was reconstructed from numerous individual observations. When the amebae were in moderately close proximity to the brush, border, the microvilli became shortened, irregular, and sometimes disappeared. Dense material was observed between the amebae and microvilli. When the ameba was very close to the epithelium the apical portion of epithelial cytoplasm projected into the lumen contacting the organism, thus becoming detached from adjoining cells. This produced spaces between epithelial cells through which amebae invaded interepithelial spaces. Initially the ameba penetrated the interglandular epithelium. Later, it penetrated equally the glandular and interglandular epithelium barrier. There were marked alterations of cytoplasmic components of epithelial cells. Polymorphonuclear leukocytes migrated into the epithelium filling these spaces; these often showed a variety of degenerative processes. Amebae, utilizing their pseudopodia, moved further through the intercellular spaces and reached the lamina propria.

Eggs of Trichuris muris hatched when implanted directly into the ceca of mice. The hatched larvae had penetrated the epithelium 40 min after introduction of the eggs. It is certain that the larvae penetrated the epithelium lining the cecal glands after entering the glands. Since penetration through the surface or luminal epithelium was not observed, it may be that the larvae enter gland openings selectively before penetrating epithelial cells.

Five mule deer fawns (Odocoileus hemionus) ranging in age from 3 to 6 weeks were given sporulated Eimeria mccordocki oocysts orally. Four of the five fawns developed coccidiosis. Initial clinical signs appeared by 8 to 9 days postinoculation and included elevated body temperature and bloody diarrhea. Dehydration and limited emaciation occurred as the disease progressed. The disease was allowed to run its course in one fawn and oocysts were passed in the feces on the 16th day following inoculation. Oocysts passage continued until the 27th day, at which time the animal appeared to be completely recovered. The remaining animals demonstrated all signs of coccidiosis and were euthanized prior to passing oocysts. Intracellular stages of E. mccordocki in these animals were confined to the last 75 to 100 cm of the ileum. The cytoplasm of each infected epithelial cell contained one to five meronts. The intestinal epithelium was inflammatory and contained many necrotic foci.

This paper reviews the major steps in alimentation, digestion, and absorption, which must be intact as a basis for normal nutrition, and discusses evidence relating parasitic infection in humans to effects on intestinal physiology. Parasites, with their ability to induce systemic toxicity and fever, to release active and toxic substances into the intestinal lumen, to compete for certain nutrients, to cause both functional and structural changes in the intestinal mucosa, and to stimulate hypermotility, which lessens the time available for digestion and absorption, can influence the alimentary process a almost every one of its steps. However, parasitic infection is likely to exert its most important impact at the very first step of the alimentary process, by adversely affecting the intake of food through any of a variety of mechanisms.

Human vaginal epithelial cells (VECs) from vaginal swabs obtained from normal women or from patients with trichomoniasis were purified, and VEC parasitism by Trichomonas vaginalis was examined. Trichomonads bound equally well to live or dead VECs, and up to 20% of VECs were parasitized. Trichomonal cytadherence of human VECs was time, temperature, and pH dependent. Saturation binding levels of live trichomonads to VECs gave approximately 2 organisms adherent to parasitized VEC. No differences in cytadherence levels were detected by different isolates to VECs from the same patient compared with adherence to VECs from normal individuals. Trypsinized, live T. vaginalis organisms failed to recognize VECs. A ligand assay identified four adhesin candidates, and only organisms without a prominent immunogen on the surface (negative phenotype) cytadhered to VECs and synthesized the adhesins, confirming the results of a recently published report by us on adherence to HeLa cell monolayers (J. F. Alderete and G. E. Garza, Infect. Immun. 56:28-33, 1988). These data show the ability of T. vaginalis to parasitize human vaginal epithelial cells in a specific receptor-ligand manner.

Eimeria cernae is recorded for the first time in England and the life-cycle is described in experimentally infected bank-voles (Clethrionomys glareolus). The pre-patent period was 6 days and the patent period was 4-6 days. Oocysts were ellipsoidal in shape and measured 20.2 x 15.9 micrometers. Sporocysts, measuring 11.5 micrometers long and 6.8 micrometers wide, possessed a small stieda body and contained a mass of granular sporocyst residuum. The endogenous stages developed in the epithelial cells of the colon and rectum. Three generations of schizonts were found. The 1st-generation schizont seen at 48 h post-infection (p.i.) contained up to 8 merozoites, the 2nd-generation schizont seen at 72 hr p.i. had a mean number of 16 (12-20) merozoites and the 3rd-generation schizont at 96 h p.i. had a mean of 18 (14-21) merozoites. Gamogonic stages were present from 96 to 120 h p.i. in the rectum only.

Germ-free and conventional chickens were infected with Lactobacillus acidophilus and/or Eimeria brunetti. Germ-free chickens acquiring single infections of E. brunetti were found to have lifecycle stages of the coccidial organism in the bursa of Fabricius.

Scanning electron micrographs showed that both epimastigotes and metacyclic trypomastigotes of Trypanosoma cruzi are attached by the flagellum to the epithelium of the rectal gland of Triatoma dimidiata. The flagellates tended to cover the surface of the gland and there was a marked predominance of epimastigotes with a round posterior end. Reproduction and metacyclogenesis seem to take place in situ, the latter apparently by twisting and elongation of the epimastigotes. Metatrypomastigotes remain attached for some time, probably by a weaker mechanism which easily allows them to loosen, facilitating expulsion with the urine or feces.

Developmental stages of Cryptosporidium sp. (Protozoa: Cryptosporiidae) were observed in tracheal epithelium of two groups of turkey poults from a farm in central Saskatchewan. Lesions associated with the parasite included excess mucus, epithelial hyperplasia and metaplasia and necrosis, and macrophage and heterophil infiltration in thickened lamina propria. Infection with the parasite was confirmed by histologic and ultrastructural examination of tracheal tissues.

The fine structure of the sexual stages of Cyclospora caryolytica and Eimeria goussevi from the epithelial cells of the small intestine of the European mole was studied by means of transmission electron microscopy. Special interest was given to the intranuclear development of C. caryolytica .

Dead and dying cells were localized by light microscopy in the mucosal epithelium of the intestine of an outbred strain (CD1) and an inbred strain (B10A) of mice by vital staining with the dye, trypan blue. In whole mounts of the intestinal wall, trails, or variable-sized clusters of blue-stained cells were seen throughout the course of infection and in mice given a range of inoculum levels. In CD1 mice, irregular trails of dead cells were seen in the intestine floor and clusters of them along the villi. In B10A mice, dead cells were seen only as trails or clusters in the intestinal floor. The results suggest that worms move through the epithelium only in the intestinal floor. Cells killed by this activity may be sloughed from the epithelium more rapidly by B10A mice than by CD1 mice where the dead cells migrate up villi before being sloughed.

The in vitro interaction between Treponema pallidum and Sf1Ep cells during treponemal replication was investigated by using transmission electron microscopy. The Sf1Ep cells grown on Teflon-treated cover slips after 12 days of cocultivation were fixed in situ, overlaid with agar, embedded, and vertically sectioned. Large numbers of treponemes were found extracellularly not only at the upper cell surfaces but also in the narrow spaces between the cells and between the cells and the cover slips. These narrow spaces supported treponemal growth and survival, as did those at the upper cell surfaces. Although few in number, organisms were also seen in cell vacuoles either surrounded by a membrane or free in the cytoplasm. Some extracellular treponemes attached to host cells by body spirals or the terminal end and formed electron-dense layers at attachment sites. Some treponemes were often surrounded with amorphous, extracellular material which appeared to "connect" them to host cell surface. After 12 days of cocultivation, host cells showed excessive vacuolation and appeared to be damaged. This did not seem to be due to treponemal infection alone, because cells from uninfected cultures also showed similar vacuolation.

Three calves were studied in stages of spontaneous cryptosporidial infection with particular reference to the relation of the cryptosporidia to the follicle-associated epithelium (FAE) over the ileal Peyer's patch (IPP). In early infection scanning electron microscopy and streptavidin immunoperoxidase staining showed marked predilection of cryptosporidia for the FAE. Cryptosporidial antigen was also found in subepithelial tissue, both in the domes over the IPP and in villi, apparently in macrophages, where the parasites seemed to be progressively degraded. The FAE showed long tightly spaced microvilli, replacing normal low folds and protrusions, particularly in late infection. Endocytosis of indian ink was restricted to the cell periphery in late infection, contrasting the normal, more even distribution of endocytosis in the FAE apical cytoplasm. Few parasites were seen in the intestinal mucosa at this stage. At convalescence the FAE was normal, but all stages of infection were characterised by elongation of microvilli in absorptive cells.

First generation merozoites of Eimeria necatrix were found in epithelial cells of the crypts of Lieberkühn of chicks cells appeared to migrate into the lamina propria, forming nests of second generation schizonts that extended from the external muscularis to the lamina propria of the mid-villus. On the fourth and fifth days the mature schizonts, usually still within the infected cells were dishcarged into adjacent crypts. The muscularis mucosa appeared to be repaired by the formation of new, smooth muscle cells that spliced the broken ends of the ruptured muscle. 'Ghosts' of schizonts were found in the lamina propria on the sixth day after infection.

In Trichostrongylus colubriformis-infected rabbits, epithelial cell migration rates and cell transit times along the villi were compared by radioautography on histological slides to normal values from noninfected small intestine. Regions of gut with high (upper jejunum) and low (ileum) burdens of worms were both examined. In the control rabbits, the estimated values for the cell migration rates in the proximal and distal parts of gut were respectively 5.8 and 2.8 microns/hr. Seventy-two hours after the thymidine injection, the labeled epithelial cells were near the tip of the villi in the jejunum whereas only 60% of the villous length was labeled in the ileum. In the infected rabbits, the presence of T. colubriformis was associated with a two-fold increase of the cell velocity, in the main site of infection. Although less prominent than in the proximal region, a significant acceleration in the cell migration was also noticed in the ileum. The cell transit time was markedly reduced in the parasitized jejunum, but no variation of this parameter was found in the distal part of gut. These changes in the dynamics of epithelial cells in both regions of the gut appeared to underlie the morphological and enzymological changes of the parasitized mucosa. They particularly contribute to create an adaptive region in the small intestine beyond the main site of infection.

Colonic epithelia from rats infected with the nematode Nippostrongylus brasiliensis have been studied under short circuit conditions and in response to challenge with worm antigen. Challenge from the serosal but not the mucosal side with antigen caused a transient increase in inwardly directed short circuit current. No effects were observed in comparable tissues from noninfected animals. Simultaneous measurements of short circuit current and of the fluxes of sodium or chloride ions showed there was an increase in electrogenic chloride secretion and an inhibition of electroneutral sodium chloride absorption, associated with antigen challenge. This result, together with the inhibitory effects of piretanide on the response to antigen challenge, indicate that chloride ions are a major carrier of the short circuit current response. However, the equivalence of the biophysical response to ion fluxes was not established, there being an excess of chloride secretion. The mast cell stabilizing agent, FPL 52694, significantly inhibited the current responses to antigen, while cromoglycate and doxantrazole were ineffective. Mepyramine, an H1-receptor antagonist, and indomethacin, an inhibitor of fatty acid cyclo-oxygenase, were without effect on the responses to antigen challenge. Anti-rat IgE produced qualitatively similar responses to antigen in both normal and sensitized colonic epithelia. However, the responses were significantly greater in tissues derived from infected animals. Maximally effective antigen concentrations prevented subsequent responses to anti-rat IgE in sensitized tissues, while anti-rat IgE only attenuated the responses to antigen. The ways in which antigen challenge modifies epithelial function is discussed, particularly in relation to its possible role in promoting rejection of the nematodes during secondary infection.

Life cycle of Eimeria coecicola was studied in experimentally infected rabbits by light microscopy and by transmission and scanning electron microscopy. First and second generation meronts developed in the vermiform appendix; third and fourth generation meronts were located in the epithelium of the ileum. Gametogony developed again in the vermiform appendix. The prepatent period was 9 days. New data were obtained by the study of asexual reproduction. First generation meronts were first observed 4 days post infection (DPI), which is relatively late in comparison with other species of rabbit coccidia. Sporozoites were found in lymphatic follicles of the vermiform appendix at 4 DPI by transmission electron microscopy. This suggests, together with selective location of first generation meronts in the epithelium adjacent to these follicles, that major part of sporozoites enter the epithelium cells through lymphatic follicles and not through the lumen of the vermiform appendix. The process of development of first generation merozoites is similar to endodyogeny. The differences are in formation of apical parts of daughter merozoites which is not coincidental with nuclear division and in formation of the outer membrane of pellicle which arises within the mother cell. Some first generation merozoites have 2-3 nuclei, second and fourth generation merozoites are only uninucleate, while third generation merozoites are only multinucleate. We found that further merozoites are formed in multinucleate third generation merozoites by endopolygeny.

During feeding a peritrophic membrane (PM) is formed in the gut of the tick Ixodes dammini, dividing the lumen of the gut into an ecto- and endoperitrophic space. Babesia and all food particles ingested with the blood meal by the tick are retained in the endoperitrophic space, the lumen proper. Only Babesia equipped with a highly specialized organelle, the arrowhead, are able to pass the PM and enter the ectoperitrophic compartment. During the crossing of the PM the arrowhead loses its density, suggesting that enzymes released from it dissolve the polymers in the PM, making passage of the parasite through this barrier possible. In the ectoperitrophic space the arrowhead of Babesia touches the epithelial cell. At the point of contact the membrane of the host cell starts to invaginate, and simultaneously the arrowhead's fine structure loses its highly organized pattern. The growing host membrane encircles the parasite and the arrowhead diminishes progressively in size. When the piroplasm is inside the host cell, the arrowhead can no longer be found. During invasion the host membrane often touches the parasite's plasma membrane at the site of a coiled structure, and the host membrane becomes ruptured and the nearby host cytoplasm appears to be lysed. Babesia inside the host cell is covered solely by its own plasma membrane; the invaginated host membrane is missing. It is postulated that the latter disintegrates during invasion by the parasite through the action of enzymes from the coiled structure. The parasite is surrounded by a halo of homogeneous material deriving most probably from the lysed host cytoplasm.

Histologic examination of the tongue of one juvenile male raccoon demonstrated the presence of adults and unembryonated eggs of the nematode Capillaria procyonis within parasite-induced tunnels in the stratified squamous epithelium. There was no inflammatory infiltrate within the epithelium or the subjacent lamina propria. The shape of tunnels approximated that of the resident nematodes and were lined by flattened and attenuated epithelial cells. Therefore, the site of infection of C. procyonis in the raccoon includes the oral, lingual and pharyngeal epithelium in addition to the esophagus.

The ultrastructure of microgametogenesis of Eimeria bakuensis was studied in the intestinal epithelium of an experimentally infected sheep. During development the nuclei became situated peripherally in the microgamont. Nuclear chromatin condensed into peripheral, dense portions that became the nuclei of the microgametes. The young microgamont was surrounded by a very narrow parasitophorous vacuole which widened during development and contained a few intravacuolar folds. Fissures and invaginations developed in the microgamont at the time the flagella began to grow. Polysaccharide granules were not present in the early and intermediate microgamonts but increased in number during the later stages and remained in the residual cytoplasm of the mature stage.

A fibrous material surrounds cells of Trypanosoma (Megatrypanum) melophagium in the hind-gut of the sheep ked, Melophagus ovinus, and terminates just beyond the distal portions of the attached cells. The fibres of this extracellular matrix have a diameter of approximately 4 nm and are closely packed. Individual fibres have approximately the same orientation as adjacent fibres and usually lie parallel to the longitudinal axis of the parasite cells.

Intestinal infection by the coccidian parasite Cryptosporidium is a well-recognized condition in immunocompromised hosts and in some normal persons. The authors studied a patient with acquired immunodeficiency syndrome and cryptosporidiosis of the small intestine. The parasite inhabits the microvillous brush border of the intestinal epithelium and must be carefully sought on light microscopic examination of intestinal biopsy specimens. Characteristic life cycle stages are observed on electron microscopy. The absence of significant light microscopic alterations of the villous architecture in this patient's biopsy specimen and in other cases suggests that other factors, such as toxin elaboration by cryptosporidia or other organisms, may be involved in the pathogenesis of diarrhea. Abnormal aggregation of lysosomes at the apices of intestinal epithelial cells may reflect ineffective host phagocytic mechanisms.

Previous observations demonstrated severe thymocyte depletion in mice undergoing acute Chagas' disease. These data led us to investigate the status of the thymic microenvironment in these animals. Young adult C57BL/6 and C3H/HeJ mice were infected i.p. with 10(5) blood-derived trypomastigote forms of Trypanosoma cruzi (CL strain) and killed 7-14 days after infection. Sera were then analyzed for thymic hormone (thymulin) levels, and frozen thymus sections were studied by immunohistochemistry for the expression of functional antigens (thymulin and Ia), the distribution of distinct thymic epithelial cell subsets and extracellular matrix components. Infected mice exhibited a transient decrease in thymulin production and those with severe thymic atrophy showed a denser Ia-bearing cellular network. In addition, an abnormal localization of the TR5 and CK18 antigens restricted to the medullary and cortical TEC subsets, respectively, was observed. Furthermore, an increase in the basement membrane proteins was detected within thymic lobules. We suggest that the thymic microenvironment is also affected during T. cruzi infection, extending the concept that the thymus should be regarded as a target in Chagas' disease.

Elevated corneal epithelial lines are another clinical sign in Acanthamoeba corneal infection. In this report, one patient wore extended wear soft contact lenses, and another wore daily wear soft contact lenses. Both patients used distilled water and salt tablets in their lens care. Histopathologic examination of these lines revealed trophozoites and cysts. In one of the patients following penetrating keratoplasty, Acanthamoeba castellani and Acanthamoeba polyphaga were cultured by impression cytology of an epithelial line, as well as from the bulbar and tarsal conjunctiva. In the other patient who did not undergo penetrating keratoplasty, these lines appeared in the cornea one month after initial symptoms.

Clinical coccidiosis in swine is associated with Isospora suis infection of piglets. The endogenous life cycle of I suis in piglets occurs within the superficial epithelium of the small intestine. Extraintestinal stages of I suis were not demonstrated in tissues of experimentally infected piglets or mice. Feeding these tissues to previously uninfected piglets did not result in oocyst shedding of consequence. Extraintestinal stages do not seem to occur in piglets with I suis as in some other Isospora species.