scholarly journals Unique Tubulin-Based Structures in the Zoonotic Apicomplexan Parasite Cryptosporidium parvum

2021 ◽  
Vol 9 (9) ◽  
pp. 1921
Author(s):  
Chenchen Wang ◽  
Dongqiang Wang ◽  
Jiawen Nie ◽  
Xin Gao ◽  
Jigang Yin ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Cryptosporidium Parvum ◽  
Polyclonal Antibody ◽  
Developmental Stages ◽  
Apicomplexan Parasite ◽  
Parasite Development ◽  
Asexual Development ◽  
Intracellular Parasite ◽  
Cytoskeletal Elements ◽  
Β Tubulin

Cryptosporidium parasites are known to be highly divergent from other apicomplexan species at evolutionary and biological levels. Here we provide evidence showing that the zoonotic Cryptosporidium parvum also differs from other apicomplexans, such as Toxoplasma gondii, by possessing only two tubulin-based filamentous structures, rather than an array of subpellicular microtubules. Using an affinity-purified polyclonal antibody against C. parvum β-tubulin (CpTubB), we observed a long and a short microtubule that are rigid and stable in the sporozoites and restructured during the intracellular parasite development. In asexual development (merogony), the two restructuring microtubules are present in pairs (one pair per nucleus or merozoites). In sexual developmental stages, tubulin-based structures are detectable only in microgametes, but undetectable in macrogametes. These observations indicate that C. parvum parasites use unique microtubule structures that differ from other apicomplexans as part of their cytoskeletal elements.

scholarly journals Functional characterization of a fatty acyl-CoA-binding protein (ACBP) from the apicomplexan Cryptosporidium parvum

Microbiology ◽  
2006 ◽  
Vol 152 (8) ◽  
pp. 2355-2363 ◽  
Author(s):  
Bin Zeng ◽  
Xiaomin Cai ◽  
Guan Zhu
Keyword(s):  
Functional Analysis ◽  
Cryptosporidium Parvum ◽  
Developmental Stages ◽  
Binding Protein ◽  
Parasitophorous Vacuole ◽  
Functional Characterization ◽  
Fatty Acyl ◽  
Repeat Sequence ◽  
Parasite Development ◽  
Protein Fusion

In this paper, the identification and functional analysis of a fatty acyl-CoA-binding protein (ACBP) gene from the opportunistic protist Cryptosporidium parvum are described. The CpACBP1 gene encodes a protein of 268 aa that is three times larger than typical ACBPs (i.e. ∼90 aa) of humans and animals. Sequence analysis indicated that the CpACBP1 protein consists of an N-terminal ACBP domain (∼90 aa) and a C-terminal ankyrin repeat sequence (∼170 aa). The entire CpACBP1 ORF was engineered into a maltose-binding protein fusion system and expressed as a recombinant protein for functional analysis. Acyl-CoA-binding assays clearly revealed that the preferred binding substrate for CpACBP1 is palmitoyl-CoA. RT-PCR, Western blotting and immunolabelling analyses clearly showed that the CpACBP1 gene is mainly expressed during the intracellular developmental stages and that the level increases during parasite development. Immunofluorescence microscopy showed that CpACBP1 is associated with the parasitophorous vacuole membrane (PVM), which implies that this protein may be involved in lipid remodelling in the PVM, or in the transport of fatty acids across the membrane.

scholarly journals Diferensiasi Stadium Takizoit-Bradizoit pada Toxoplasma gondii

2017 ◽  
Vol 4 (2) ◽  
pp. 89
Author(s):  
Suri Dwi Lesmana
Keyword(s):  
Toxoplasma Gondii ◽  
Intermediate Host ◽  
Interferon Gamma ◽  
Life Stage ◽  
Apicomplexan Parasite ◽  
Intracellular Parasite ◽  
Symptomatic Disease ◽  
Definite Host ◽  
Differentiation Event ◽  
Bradyzoite Differentiation

Toxoplasma gondii is an intracellular parasite whose definite host is cat or felidae and intermediate host is human orother mammals. It causes congenital and acquisita toxoplasmosis. Infection with this apicomplexan parasite results inits dissemination throughout its host via the tachyzoite life stage. After dissemination, these tachyzoites differentiateinto bradyzoites within cyst and remain latent. These bradyzoites can transform back into tachyzoites and inimmunosupressed individuals this often results in symptomatic disease. Both tachyzoites and bradyzoites develop intissue culture and this crucial differentiation event can be studied. Interferon gamma (IFN ) is the main mediator intachyzoite-bradyzoite differentiation.

Ultrastructural and molecular characterization of Bacillidium vesiculoformis n. sp. (Microspora: Mrazekiidae) in the freshwater oligochaete Nais simplex (Oligochaeta: Naididae)

Parasitology ◽  
2004 ◽  
Vol 130 (1) ◽  
pp. 31-40 ◽  
Author(s):  
D. J. MORRIS ◽  
R. S. TERRY ◽  
K. B. FERGUSON ◽  
J. E. SMITH ◽  
A. ADAMS
Keyword(s):  
Phylogenetic Analysis ◽  
New Species ◽  
Infected Cell ◽  
Host Cell ◽  
Developmental Stages ◽  
Parasite Development ◽  
Adhesive Quality ◽  
Transmission Studies ◽  
A New Species

The development of a new species, Bacillidium vesiculoformis n. sp. (Microspora, Mrazekiidae), is described from the freshwater oligochaete Nais simplex (Oligochaeta, Naididae). Initial stages of parasite development consist of a monokaryotic merogony within a haemocyte of the intestinal blood sinus. The resulting hypertrophied haemocyte is attached to the chloragocytes of the sinus by fine cytoplasmic extensions with the sinus around the cell becoming greatly enlarged. The meronts within the haemocyte form diplokaryotic sporonts that undergo sporogenesis directly within the cytoplasm of the host cell. The infected cell becomes packed with spores and developmental stages, causing it dramatically to increase in size, eventually rupturing the oligochaete and cell. Sporogony appears to be disporoblastic. Released spores were observed to have an adhesive quality. Transmission studies conducted with mature spores failed to transmit the parasite horizontally although vertical transmission was observed. Phylogenetic analysis of the parasite demonstrated that B. vesiculoformis clustered with microsporidian parasites of bryozoa and two other microsporidians, Janacekia debaiseuxi and an unidentified Bacillidium sp.

scholarly journals Death Receptor Ligation or Exposure to Perforin Trigger Rapid Egress of the Intracellular Parasite Toxoplasma gondii

2007 ◽  
Vol 179 (12) ◽  
pp. 8357-8365 ◽  
Author(s):  
Emma K. Persson ◽  
Abela Mpobela Agnarson ◽  
Henrik Lambert ◽  
Niclas Hitziger ◽  
Hideo Yagita ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Death Receptor ◽  
Intracellular Parasite ◽  
Receptor Ligation

scholarly journals Distinct gene expression patterns in vector-residing Leishmania infantum identify parasite stage-enriched markers

2019 ◽  
Author(s):  
Iliano V. Coutinho-Abreu ◽  
Tiago D. Serafim ◽  
Claudio Meneses ◽  
Shaden Kamhawi ◽  
Fabiano Oliveira ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Developmental Stages ◽  
Expression Patterns ◽  
Principal Component ◽  
Sand Fly ◽  
Parasite Development ◽  
Lutzomyia Longipalpis ◽  
Specific Marker ◽  
Limited Information ◽  
Natural Sand

AbstractPromastigotes of Leishmania infantum undergo a series of extracellular developmental stages inside the natural sand fly vector Lutzomyia longipalpis to reach the infectious stage, the metacyclic promastigote. There is limited information regarding the expression profile of L. infantum developmental stages inside the sand fly vector, and molecular markers that can distinguish the different parasite stages are lacking. We performed RNAseq on unaltered midguts of the sand fly Lutzomyia longipalpis after infection with L. infantum parasites. RNAseq was carried out at various time points throughout parasite development. Principal component analysis mapped the sequences corresponding to the procyclic, nectomonad, leptomonad or metacyclic promastigote stage into distinct positions, with the procyclic stage being the most divergent population. Transcriptional levels across genes varied on average between 10- to 100-fold. Comparison between procyclic and nectomonad promastigotes resulted in 836 differentially expressed (DE) genes; between nectomonad and leptomonad promastigotes in 113 DE genes; and between leptomonad and metacyclic promastigotes in 302 DE genes. Most of the DE genes do not overlap across stages, highlighting the uniqueness of each stage. Furthermore, the different stages of Leishmania parasites exhibited specific transcriptional enrichment across chromosomes. Using the transcriptional signatures exhibited by distinct Leishmania stages during their development in the sand fly midgut, we determined the genes predominantly enriched in each stage, identifying multiple stage-specific markers for L. Infantum. Leading stage-specific marker candidates include genes encoding a zinc transporter in procyclics, a beta-fructofuranidase in nectomonads, a surface antigen-like protein in leptomonads, and an amastin-like surface protein in metacyclics. Overall, these findings demonstrate the transcriptional plasticity of the Leishmania parasite inside the sand fly vector and provide a repertoire of stage-specific markers for further development as molecular tools for epidemiological studies.

scholarly journals Toxoplasma gondii Rhoptry Discharge Correlates with Activation of the Early Growth Response 2 Host Cell Transcription Factor

2008 ◽  
Vol 76 (10) ◽  
pp. 4703-4712 ◽  
Author(s):  
Eric D. Phelps ◽  
Kristin R. Sweeney ◽  
Ira J. Blader
Keyword(s):  
Transcription Factor ◽  
Toxoplasma Gondii ◽  
Host Cell ◽  
Growth Response ◽  
Neospora Caninum ◽  
Early Growth ◽  
Apicomplexan Parasite ◽  
Host Cells ◽  
Early Growth Response ◽  
Cell Extracts

ABSTRACT Toxoplasma gondii is a ubiquitous apicomplexan parasite that can cause severe disease in fetuses and immune-compromised patients. Rhoptries, micronemes, and dense granules, which are secretory organelles unique to Toxoplasma and other apicomplexan parasites, play critical roles in parasite growth and virulence. To understand how these organelles modulate infected host cells, we sought to identify host cell transcription factors triggered by their release. Early growth response 2 (EGR2) is a host cell transcription factor that is rapidly upregulated and activated in Toxoplasma-infected cells but not in cells infected with the closely related apicomplexan parasite Neospora caninum. EGR2 upregulation occurred only when live parasites were in direct contact with the host cell and not from exposure to cell extracts that contain dense granule or micronemal proteins. When microneme-mediated attachment was blocked by pretreating parasites with a calcium chelator, EGR2 expression was significantly reduced. In contrast, when host cells were infected with parasites in the presence of cytochalasin D, which allows rhoptry secretion but prevents parasite invasion, EGR2 was activated. Finally, we demonstrate that Toxoplasma activation of host p38 mitogen-activated protein kinase is necessary but not sufficient for EGR2 activation. Collectively, these data indicate that EGR2 is specifically upregulated by a parasite-derived secreted factor that is most likely a resident rhoptry protein.

Unique cultural methods used to detect viable cryptosporidium parvum oocysts in environmental samples

1997 ◽  
Vol 35 (11-12) ◽  
pp. 363-368 ◽  
Author(s):  
T. R. Slifko ◽  
D. E. Friedman ◽  
J. B. Rose ◽  
S. J. Upton ◽  
W. Jakubowski
Keyword(s):  
Cryptosporidium Parvum ◽  
Environmental Samples ◽  
Developmental Stages ◽  
High Sensitivity ◽  
Protozoan Parasite ◽  
Culture Methods ◽  
Low Level ◽  
Cultural Methods ◽  
Severe Gastroenteritis ◽  
Cell Culture Methods

Cryptosporidium parvum is an infectious enteric protozoan parasite that causes waterborne disease, severe gastroenteritis and is associated with high mortality in immunocompromised individuals. Detection of oocysts in water is very difficult and current methodologies do not determine viability. This project has focused on low level detection of Cryptosporidium parvum in environmental samples using a unique cultural method. Previously, cell culture methods have been used to assess the developmental stages of Cryptosporidium; however, no cultural methods have been employed with environmental samples. The percentage of viable oocysts can be estimated by detecting intracellular developmental stages of the parasite using fluorescently labelled antibodies. Other methods are not capable of low level detection or high sensitivity. We are evaluating detection of single foci of infection, indicating that one of the four sporozoites released from the viable oocyst has infected a single cell.

scholarly journals Anopheles stephensi Dual Oxidase Silencing Activates the Thioester-Containing Protein 1 Pathway to Suppress Plasmodium Development

2019 ◽  
Vol 11 (6) ◽  
pp. 496-505 ◽  
Author(s):  
Parik Kakani ◽  
Mithilesh Kajla ◽  
Tania Pal Choudhury ◽  
Lalita Gupta ◽  
Sanjeev Kumar
Keyword(s):  
Amino Acid ◽  
Calcium Binding ◽  
Anopheles Stephensi ◽  
Developmental Stages ◽  
Transmembrane Protein ◽  
Phylogenetic Analyses ◽  
Amino Acid Identity ◽  
Infected Mosquito ◽  
Parasite Development ◽  
Dual Oxidase

We characterized the dual oxidase (Duox) gene in the major Indian malaria vector Anopheles stephensi, which regulates the generation of reactive oxygen species. The AsDuox gene encodes for a 1,475-amino-acid transmembrane protein that contains an N-terminal noncytoplasmic heme peroxidase domain, a calcium-binding domain, seven transmembrane domains, and a C-terminal cytoplasmic NADPH domain. Phylogenetic analyses revealed that A. stephensi Duox protein is highly conserved and shares 97–100% amino acid identity with other anopheline Duoxes. AsDuox is expressed in all the developmental stages of A. stephensi and the pupal stages revealed relatively higher expressions. The Duox gene is induced in Plasmodium-infected mosquito midguts, and RNA interference-mediated silencing of this gene suppressed parasite development through activation of the thioester-containing protein 1 pathway. We propose that this highly conserved anopheline Duox, being a Plasmodium agonist, is an excellent target to control malaria parasite development inside the insect host.

scholarly journals A selector of transcription initiation in the protozoan parasite Toxoplasma gondii.

1995 ◽  
Vol 15 (1) ◽  
pp. 87-93 ◽  
Author(s):  
D Soldati ◽  
J C Boothroyd
Keyword(s):  
Toxoplasma Gondii ◽  
Gene Transcription ◽  
Transcription Initiation ◽  
Protozoan Parasite ◽  
Intracellular Parasite ◽  
Transcription Start ◽  
Transcription Start Sites ◽  
Obligate Intracellular ◽  
Obligate Intracellular Parasite ◽  
Sag1 Gene

The recent development of an efficient transfection system for the apicomplexan Toxoplasma gondii allows a comprehensive dissection of the elements involved in gene transcription in this obligate intracellular parasite. We demonstrate here that for the SAG1 gene, a stretch of six repeated sequences in the region 35 to 190 bp upstream of the first of two transcription start sites is essential for efficient and accurate transcription initiation. This repeat element shows characteristics of a selector in determining the position of the transcription start sites.

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