Cryptosporidium parvum: Structural Components of the Oocyst Wall

1999 ◽  
Vol 85 (5) ◽  
pp. 839 ◽  
Author(s):  
J. Robin Harris ◽  
Franz Petry
Keyword(s):  
Cryptosporidium Parvum ◽  
Structural Components ◽  
Oocyst Wall

scholarly journals Strategies To Discover the Structural Components of Cyst and Oocyst Walls

2013 ◽  
Vol 12 (12) ◽  
pp. 1578-1587 ◽  
Author(s):  
John Samuelson ◽  
G. Guy Bushkin ◽  
Aparajita Chatterjee ◽  
Phillips W. Robbins
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibodies ◽  
Cryptosporidium Parvum ◽  
Giardia Lamblia ◽  
Intestinal Lumen ◽  
Structural Components ◽  
Oocyst Wall ◽  
Content Type ◽  
Glucose Polymer ◽  
Polymer Models

ABSTRACTCysts ofGiardia lambliaandEntamoeba histolyticaand oocysts ofToxoplasma gondiiandCryptosporidium parvumare the infectious and sometimes diagnostic forms of these parasites. To discover the structural components of cyst and oocyst walls, we have developed strategies based upon a few simple assumptions. Briefly, the most abundant wall proteins are identified by monoclonal antibodies or mass spectrometry. Structural components include a sugar polysaccharide (chitin forEntamoeba, β-1,3-linked glucose forToxoplasma, and β-1,3-linked GalNAc forGiardia) and/or acid-fast lipids (ToxoplasmaandCryptosporidium). BecauseEntamoebacysts andToxoplasmaoocysts are difficult to obtain, studies of walls of nonhuman pathogens (E. invadensandEimeria, respectively) accelerate discovery. Biochemical methods to dissect fungal walls work well for cyst and oocyst walls, although the results are often unexpected. For example, echinocandins, which inhibit glucan synthases and kill fungi, arrest the development of oocyst walls and block their release into the intestinal lumen.Candidawalls are coated with mannans, whileEntamoebacysts are coated in a dextran-like glucose polymer. Models for cyst and oocyst walls derive from their structural components and organization within the wall. Cyst walls are composed of chitin fibrils and lectins that bind chitin (Entamoeba) or fibrils of the β-1,3-GalNAc polymer and lectins that bind the polymer (Giardia). Oocyst walls ofToxoplasmahave two distinct layers that resemble those of fungi (β-1,3-glucan in the inner layer) or mycobacteria (acid-fast lipids in the outer layer). Oocyst walls ofCryptosporidiumhave a rigid bilayer of acid-fast lipids and inner layer of oocyst wall proteins.

scholarly journals Silver Nanoparticles Decrease the Viability of Cryptosporidium parvum Oocysts

2015 ◽  
Vol 82 (2) ◽  
pp. 431-437 ◽  
Author(s):  
Pamela Cameron ◽  
Birgit K. Gaiser ◽  
Bidha Bhandari ◽  
Paul M. Bartley ◽  
Frank Katzer ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Cryptosporidium Parvum ◽  
Silver Ions ◽  
Protozoan Parasite ◽  
Dependent Manner ◽  
Oocyst Wall ◽  
Content Type ◽  
Chlorine Disinfection ◽  
High Concentrations ◽  
Dose Dependent

ABSTRACTOocysts of the waterborne protozoan parasiteCryptosporidium parvumare highly resistant to chlorine disinfection. We show here that both silver nanoparticles (AgNPs) and silver ions significantly decrease oocyst viability, in a dose-dependent manner, between concentrations of 0.005 and 500 μg/ml, as assessed by an excystation assay and the shell/sporozoite ratio. For percent excystation, the results are statistically significant for 500 μg/ml of AgNPs, with reductions from 83% for the control to 33% with AgNPs. For Ag ions, the results were statistically significant at 500 and 5,000 μg/ml, but the percent excystation values were reduced only to 66 and 62%, respectively, from 86% for the control. The sporozoite/shell ratio was affected to a greater extent following AgNP exposure, presumably because sporozoites are destroyed by interaction with NPs. We also demonstrated via hyperspectral imaging that there is a dual mode of interaction, with Ag ions entering the oocyst and destroying the sporozoites while AgNPs interact with the cell wall and, at high concentrations, are able to fully break the oocyst wall.

Characterization and immunolocalization of an oocyst wall antigen ofCryptosporidium parvum(Protozoa: Apicomplexa)

Parasitology ◽  
1991 ◽  
Vol 103 (2) ◽  
pp. 171-177 ◽  
Author(s):  
A. Bonnin ◽  
J. F. Dubremetz ◽  
P. Camerlynck
Keyword(s):  
Monoclonal Antibody ◽  
Immunoelectron Microscopy ◽  
Cryptosporidium Parvum ◽  
Parasitophorous Vacuole ◽  
Periodate Oxidation ◽  
Immunofluorescence Assay ◽  
Minor Components ◽  
Oocyst Wall ◽  
Gold Particles ◽  
Electron Lucent

A monoclonal antibody (OW-IGO) raised against purified excysted oocysts ofCryptosporidium parvumreacted in an immunofluorescence assay with the oocyst wall. The corresponding antigen was localized by immunoelectron microscopy in fibrillous material present in the parasitophorous vacuole of developing macrogametes and in the wall of both single and double layered sporulating oocysts. Gold particles were also detected over electron-lucent vesicles of the macrogametes by immunoelectron microscopy. On Western blotting ofC. parvumoocyst extracts, major bands at 250 and 40 kDa and several minor components were recognized by Mab OW-IGO. Almost complete abolition of Western blot reactivity occurred after periodate oxidation of oocyst antigen, suggesting that monoclonal antibody OW-IGO reacts with a carbohydrate epitope. Taken together, our results suggest that a fibrillous glycoproteinic material is released in the parasitophorous vacuole from electron-lucent vesicles during gametogenesis, and later condensed in the oocyst wall.

scholarly journals Cloning and Expression of a DNA Sequence Encoding a 41-Kilodalton Cryptosporidium parvum Oocyst Wall Protein

1999 ◽  
Vol 6 (6) ◽  
pp. 912-920 ◽  
Author(s):  
Mark C. Jenkins ◽  
Jim Trout ◽  
Charles Murphy ◽  
James A. Harp ◽  
Jim Higgins ◽  
...  
Keyword(s):  
Cryptosporidium Parvum ◽  
Recombinant Dna ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cloning And Expression ◽  
Wall Material ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Oocyst Wall ◽  
Species Specific

ABSTRACT This study was conducted to produce a recombinant species-specific oocyst wall protein of Cryptosporidium parvum. Antigens unique to C. parvum were identified by gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of oocyst proteins from several different Cryptosporidiumspecies. Antiserum was then prepared against a 41-kDa antigen unique toC. parvum and used to identify a recombinant DNA clone, designated rCP41. Expression of CP41 mRNA in C. parvumoocysts was confirmed by reverse transcriptase PCR (RT-PCR). Although the CP41 sequence was shown by PCR to be present in the genome ofC. baileyi, CP41 mRNA was not detected in this species by RT-PCR. Immunofluorescence staining with antiserum against recombinant CP41 detected native CP41 antigen on the surface of C. parvum oocysts but failed to detect CP41 on C. baileyi oocysts. Immunoelectron microscopy demonstrated that native CP41 was distributed unevenly on the C. parvumoocyst surface and was associated with amorphous oocyst wall material. In an enzyme-linked immunosorbent assay, purified rCP41 performed as well as native C. parvum oocyst protein in measuring the serological responses of young calves and adult cows to experimental and natural C. parvum infections. These results indicate that recombinant CP41 antigen may have potential in the immunodiagnosis of cryptosporidiosis.

Composition and Conformation of Cryptosporidium parvum Oocyst Wall Surface Macromolecules and Their Effect on Adhesion Kinetics of Oocysts on Quartz Surface

2010 ◽  
Vol 11 (8) ◽  
pp. 2109-2115 ◽  
Author(s):  
Yuanyuan Liu ◽  
Mark S. Kuhlenschmidt ◽  
Theresa B. Kuhlenschmidt ◽  
Thanh H. Nguyen
Keyword(s):  
Cryptosporidium Parvum ◽  
Quartz Surface ◽  
Wall Surface ◽  
Oocyst Wall ◽  
Cryptosporidium Parvum Oocyst ◽  
Kinetics Of

scholarly journals Genetic analysis of Cryptosporidium from 2414 humans with diarrhoea in England between 1985 and 2000

2006 ◽  
Vol 55 (6) ◽  
pp. 703-707 ◽  
Author(s):  
F. Leoni ◽  
C. Amar ◽  
G. Nichols ◽  
S. Pedraza-Díaz ◽  
J. McLauchlin
Keyword(s):  
Genetic Analysis ◽  
Cryptosporidium Parvum ◽  
18S Rdna ◽  
Oocyst Wall ◽  
Cryptosporidium Hominis ◽  
Rdna Genes ◽  
Cryptosporidium Andersoni ◽  
Faecal Samples ◽  
Pcr Rflp

The characterization of Cryptosporidium using DNA extracted from whole faecal samples collected from 2414 humans with diarrhoea in England between 1985 and 2000 where cryptosporidial oocysts were detected using conventional methods is described. Characterization was achieved by PCR/RFLP and DNA sequencing of fragments of the Cryptosporidium oocyst wall protein and the 18S rDNA genes. Cryptosporidium parvum was detected in 56.1 % of cases, Cryptosporidium hominis in 41.7 % and a mixture of C. parvum and C. hominis in 0.9 %. In the remainder of cases, Cryptosporidium meleagridis (0.9 %), Cryptosporidium felis (0.2 %), Cryptosporidium andersoni (0.1 %), Cryptosporidium canis (0.04 %), Cryptosporidium suis (0.04 %) and the Cryptosporidium cervine type (0.04 %) were detected.

scholarly journals Evidence for Mucin-Like Glycoproteins That Tether Sporozoites of Cryptosporidium parvum to the Inner Surface of the Oocyst Wall

2009 ◽  
Vol 9 (1) ◽  
pp. 84-96 ◽  
Author(s):  
Anirban Chatterjee ◽  
Sulagna Banerjee ◽  
Martin Steffen ◽  
Roberta M. O'Connor ◽  
Honorine D. Ward ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Monoclonal Antibodies ◽  
Cryptosporidium Parvum ◽  
Oral Route ◽  
Ruthenium Red ◽  
Oocyst Wall ◽  
Maclura Pomifera ◽  
Inner Surface ◽  
Ruthenium Red Staining ◽  
Binding Lectin

ABSTRACT Cryptosporidium parvum oocysts, which are spread by the fecal-oral route, have a single, multilayered wall that surrounds four sporozoites, the invasive form. The C. parvum oocyst wall is labeled by the Maclura pomifera agglutinin (MPA), which binds GalNAc, and the C. parvum wall contains at least two unique proteins ( C ryptosporidium oocyst wall protein 1 [COWP1] and COWP8) identified by monoclonal antibodies. C. parvum sporozoites have on their surface multiple mucin-like glycoproteins with Ser- and Thr-rich repeats (e.g., gp40 and gp900). Here we used ruthenium red staining and electron microscopy to demonstrate fibrils, which appear to attach or tether sporozoites to the inner surface of the C. parvum oocyst wall. When disconnected from the sporozoites, some of these fibrillar tethers appear to collapse into globules on the inner surface of oocyst walls. The most abundant proteins of purified oocyst walls, which are missing the tethers and outer veil, were COWP1, COWP6, and COWP8, while COWP2, COWP3, and COWP4 were present in trace amounts. In contrast, MPA affinity-purified glycoproteins from C. parvum oocysts, which are composed of walls and sporozoites, included previously identified mucin-like glycoproteins, a GalNAc-binding lectin, a Ser protease inhibitor, and several novel glycoproteins (C. parvum MPA affinity-purified glycoprotein 1 [CpMPA1] to CpMPA4). By immunoelectron microscopy (immuno-EM), we localized mucin-like glycoproteins (gp40 and gp900) to the ruthenium red-stained fibrils on the inner surface wall of oocysts, while antibodies to the O-linked GalNAc on glycoproteins were localized to the globules. These results suggest that mucin-like glycoproteins, which are associated with the sporozoite surface, may contribute to fibrils and/or globules that tether sporozoites to the inner surface of oocyst walls.

A comparative study on the biology of Cryptosporidium sp. from guinea pigs and Cryptosporidium parvum (Apicomplexa)

1991 ◽  
Vol 37 (12) ◽  
pp. 949-952 ◽  
Author(s):  
Michael Tilley ◽  
Steve J. Upton ◽  
Clarence E. Chrisp
Keyword(s):  
Guinea Pig ◽  
Cryptosporidium Parvum ◽  
Banding Pattern ◽  
Guinea Pigs ◽  
Size Range ◽  
Molecular Size ◽  
Shape Index ◽  
Oocyst Wall ◽  
Suckling Mice ◽  
Length Width

Cryptosporidum sp. from guinea pigs and C. parvum were compared morphologically, electrophoretically, and for the ability to infect suckling mice. Oocysts from guinea pigs measured 5.4 × 4.6 (4.8–5.6 × 4.0–5.0) μm and had a shape index (length/width) of 1.17 (1.04–1.33). Oocysts of C. parvum were similar and measured 5.2 × 4.6 (4.8–5.6 × 4.2–4.8) μm with a shape index of 1.16 (1.04–1.33). All suckling mice inoculated with oocyts of C. parvum became infected, whereas most, but not all, mice fed oocysts of the guinea pig isolate also became infected. However, mice inoculated with oocysts from guinea pigs produced on average 100-fold fewer oocysts by day 7 postinoculation than did mice infected with C. parvum, and the resulting infections were sparse and patchy along the ileum. Electrophoretic profiles were similar, but 125I surface labeling of outer oocyst wall proteins revealed striking differences between the two isolates. Cryptosporidium parvum had a wide molecular size range of 125I-labeled bands, whereas C. sp. from guinea pigs had a banding pattern clustered between 39 and 66 kDa, with a smaller number of bands >100 kDa. Key words: Cryptosporidium parvum, coccidia, Apicomplexa, guinea pig, mouse.

Ultrastructural changes in the oocyst wall during excystation of Cryptosporidium parvum (Apicomplexa; Eucoccidiorida)

1985 ◽  
Vol 63 (8) ◽  
pp. 1892-1896 ◽  
Author(s):  
David W. Reduker ◽  
C. A. Speer ◽  
John A. Blixt
Keyword(s):  
Cryptosporidium Parvum ◽  
Outer Layer ◽  
Ultrastructural Changes ◽  
Oocyst Wall ◽  
Thin Electron ◽  
Cryptosporidium Parvum Oocysts ◽  
Electron Lucent

Cryptosporidium parvum oocysts were obtained from the feces of naturally infected calves and the oocyst wall was examined ultrastructurally. The oocyst wall averaged 49.7 nm thick and was composed of two layers. The outer layer was irregular in thickness, averaging 10 nm. A thin, electron-lucent space (2.5 nm) was interposed between the outer and inner layers. The inner layer had an outer (11.6 nm) and an inner (25.8 nm) zone. A suture that extended partway around the oocyst was observed within the inner layer of the oocyst wall. The suture underwent dissolution during excystation.

scholarly journals Significance of Wall Structure, Macromolecular Composition, and Surface Polymers to the Survival and Transport of Cryptosporidium parvum Oocysts

2010 ◽  
Vol 76 (6) ◽  
pp. 1926-1934 ◽  
Author(s):  
Michael B. Jenkins ◽  
Barbara S. Eaglesham ◽  
Larry C. Anthony ◽  
Scott C. Kachlany ◽  
Dwight D. Bowman ◽  
...  
Keyword(s):  
Cryptosporidium Parvum ◽  
Blue Staining ◽  
Wall Structure ◽  
Inactivation Kinetics ◽  
Aquatic Environments ◽  
Dense Layer ◽  
Oocyst Wall ◽  
Variable Surface ◽  
Biochemical Analyses ◽  
Hydrologic Transport

ABSTRACT The structure and composition of the oocyst wall are primary factors determining the survival and hydrologic transport of Cryptosporidium parvum oocysts outside the host. Microscopic and biochemical analyses of whole oocysts and purified oocyst walls were undertaken to better understand the inactivation kinetics and hydrologic transport of oocysts in terrestrial and aquatic environments. Results of microscopy showed an outer electron-dense layer, a translucent middle layer, two inner electron-dense layers, and a suture structure embedded in the inner electron-dense layers. Freeze-substitution showed an expanded glycocalyx layer external to the outer bilayer, and Alcian Blue staining confirmed its presence on some but not all oocysts. Biochemical analyses of purified oocyst walls revealed carbohydrate components, medium- and long-chain fatty acids, and aliphatic hydrocarbons. Purified walls contained 7.5% total protein (by the Lowry assay), with five major bands in SDS-PAGE gels. Staining of purified oocyst walls with magnesium anilinonaphthalene-8-sulfonic acid indicated the presence of hydrophobic proteins. These structural and biochemical analyses support a model of the oocyst wall that is variably impermeable and resistant to many environmental pressures. The strength and flexibility of oocyst walls appear to depend on an inner layer of glycoprotein. The temperature-dependent permeability of oocyst walls may be associated with waxy hydrocarbons in the electron-translucent layer. The complex chemistry of these layers may explain the known acid-fast staining properties of oocysts, as well as some of the survival characteristics of oocysts in terrestrial and aquatic environments. The outer glycocalyx surface layer provides immunogenicity and attachment possibilities, and its ephemeral nature may explain the variable surface properties noted in oocyst hydrologic transport studies.

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