scholarly journals The Enhanced Expression of Cruzipain-Like Molecules in the Phytoflagellate Phytomonas serpens Recovered From the Invertebrate and Plant Hosts

2022 ◽  
Vol 11 ◽  
Author(s):  
Simone S. C. Oliveira ◽  
Camila G. R. Elias ◽  
Felipe A. Dias ◽  
Angela H. Lopes ◽  
Claudia M. d’Avila-Levy ◽  
...  
Keyword(s):  
Life Cycle ◽  
Tomato Fruit ◽  
Protozoan Parasite ◽  
Etiologic Agent ◽  
Oncopeltus Fasciatus ◽  
Tomato Extract ◽  
Cysteine Peptidases ◽  
Plant Hosts ◽  
Enhanced Expression ◽  
Important Virulence Factor

Phytomonas serpens is a protozoan parasite that alternates its life cycle between two hosts: an invertebrate vector and the tomato fruit. This phytoflagellate is able to synthesize proteins displaying similarity to the cysteine peptidase named cruzipain, an important virulence factor from Trypanosoma cruzi, the etiologic agent of Chagas disease. Herein, the growth of P. serpens in complex medium (BHI) supplemented with natural tomato extract (NTE) resulted in the increased expression of cysteine peptidases, as verified by the hydrolysis of the fluorogenic substrate Z-Phe-Arg-AMC and by gelatin-SDS-PAGE. Phytoflagellates showed no changes in morphology, morphometry and viability, but the proliferation was slightly reduced when cultivated in the presence of NTE. The enhanced proteolytic activity was accompanied by a significant increase in the expression of cruzipain-like molecules, as verified by flow cytometry using anti-cruzipain antibodies. In parallel, parasites incubated under chemically defined conditions (PBS supplemented with glucose) and added of different concentration of NTE revealed an augmentation in the production of cruzipain-like molecules in a typically dose-dependent way. Similarly, P. serpens recovered from the infection of mature tomatoes showed an increase in the expression of molecules homologous to cruzipain; however, cells showed a smaller size compared to parasites grown in BHI medium. Furthermore, phytoflagellates incubated with dissected salivary glands from Oncopeltus fasciatus or recovered from the hemolymph of infected insects also showed a strong enhance in the expression of cruzipain-like molecules that is more relevant in the hemolymph. Collectively, our results showed that cysteine peptidases displaying similarities to cruzipain are more expressed during the life cycle of the phytoflagellate P. serpens both in the invertebrate and plant hosts.

scholarly journals Metabolic reprogramming during the Trypanosoma brucei life cycle

F1000Research ◽  
2017 ◽  
Vol 6 ◽  
pp. 683 ◽  
Author(s):  
Terry K. Smith ◽  
Frédéric Bringaud ◽  
Derek P. Nolan ◽  
Luisa M. Figueiredo
Keyword(s):  
Life Cycle ◽  
Trypanosoma Brucei ◽  
Model Organism ◽  
Protozoan Parasite ◽  
Tsetse Fly ◽  
Metabolic Reprogramming ◽  
Mammalian Host ◽  
Insect Vector ◽  
Environmental Signals ◽  
Cellular Metabolic Activity

Cellular metabolic activity is a highly complex, dynamic, regulated process that is influenced by numerous factors, including extracellular environmental signals, nutrient availability and the physiological and developmental status of the cell. The causative agent of sleeping sickness, Trypanosoma brucei, is an exclusively extracellular protozoan parasite that encounters very different extracellular environments during its life cycle within the mammalian host and tsetse fly insect vector. In order to meet these challenges, there are significant alterations in the major energetic and metabolic pathways of these highly adaptable parasites. This review highlights some of these metabolic changes in this early divergent eukaryotic model organism.

Developmental gene expression in Leishmania donovani: differential cloning and analysis of an amastigote-stage-specific gene

1994 ◽  
Vol 14 (5) ◽  
pp. 2975-2984
Author(s):  
H Charest ◽  
G Matlashewski
Keyword(s):  
Life Cycle ◽  
Leishmania Donovani ◽  
Repetitive Sequences ◽  
Protozoan Parasite ◽  
Life Cycle Stage ◽  
Immune Serum ◽  
Sand Fly ◽  
Specific Gene ◽  
Reading Frame ◽  
Developmental Gene Expression

Leishmania protozoans are the causative agents of leishmaniasis, a major parasitic disease in humans. During their life cycle, Leishmania protozoans exist as flagellated promastigotes in the sand fly vector and as nonmotile amastigotes in the mammalian hosts. The promastigote-to-amastigote transformation occurs in the phagolysosomal compartment of the macrophage cell and is a critical step for the establishment of the infection. To study this cytodifferentiation process, we differentially screened an amastigote cDNA library with life cycle stage-specific cDNA probes and isolated seven cDNAs representing amastigote-specific transcripts. Five of these were closely related (A2 series) and recognized, by Northern (RNA) blot analyses, a 3.5-kb transcript in amastigotes and in amastigote-infected macrophages. Expression of the amastigote-specific A2 gene was induced in promastigotes when they were transferred from culture medium at 26 degrees C and pH 7.4 to medium at 37 degrees C and pH 4.5, conditions which mimic the macrophage phagolysosomal environment. A2 genes are clustered in tandem arrays, and a 6-kb fragment corresponding to a unit of the cluster was cloned and partially sequenced. An open reading frame found within the A2-transcribed region potentially encoded a 22-kDa protein containing repetitive sequences. The recombinant A2 protein produced in Escherichia coli cells was specifically recognized by immune serum from a patient with visceral leishmaniasis. The A2 protein repetitive element has strong homology with an S antigen of Plasmodium falciparum, the protozoan parasite responsible for malaria. Both the A2 protein of Leishmania donovani and the S antigen of P. falciparum are stage specific and developmentally expressed in mammalian hosts.

scholarly journals An Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in Trypanosoma cruzi

mSphere ◽  
2018 ◽  
Vol 3 (1) ◽  
Author(s):  
Teresa Cruz-Bustos ◽  
Evgeniy Potapenko ◽  
Melissa Storey ◽  
Roberto Docampo
Keyword(s):  
Life Cycle ◽  
Amino Acid ◽  
Osmotic Stress ◽  
Trypanosoma Cruzi ◽  
Etiologic Agent ◽  
Ammonium Transporter ◽  
Amino Acid Consumption ◽  
Acid Consumption ◽  
Acidic Compartments ◽  
Production Of Ammonia

Trypanosoma cruzi is an important human and animal pathogen and the etiologic agent of Chagas disease. The parasite undergoes drastic changes in its metabolism during its life cycle. Amino acid consumption becomes important in the infective stages and leads to the production of ammonia (NH3), which needs to be detoxified. We report here the identification of an ammonium (NH4 +) transporter that localizes to acidic compartments and is important for replication, differentiation, and resistance to starvation and osmotic stress.

scholarly journals Elucidating the Mechanism of Trypanosoma cruzi Acquisition by Triatomine Insects: Evidence from a Large Field Survey of Triatoma infestans

2020 ◽  
Vol 5 (2) ◽  
pp. 87
Author(s):  
Aaron W. Tustin ◽  
Ricardo Castillo-Neyra ◽  
Laura D. Tamayo ◽  
Renzo Salazar ◽  
Katty Borini-Mayorí ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Regression Models ◽  
Control Strategies ◽  
Protozoan Parasite ◽  
Etiologic Agent ◽  
Triatoma Infestans ◽  
Large Field ◽  
Blood Sucking ◽  
Triatomine Bugs ◽  
Cruzi Infection

Blood-sucking triatomine bugs transmit the protozoan parasite Trypanosoma cruzi, the etiologic agent of Chagas disease. We measured the prevalence of T. cruzi infection in 58,519 Triatoma infestans captured in residences in and near Arequipa, Peru. Among bugs from infected colonies, T. cruzi prevalence increased with stage from 12% in second instars to 36% in adults. Regression models demonstrated that the probability of parasite acquisition was roughly the same for each developmental stage. Prevalence increased by 5.9% with each additional stage. We postulate that the probability of acquiring the parasite may be related to the number of feeding events. Transmission of the parasite does not appear to be correlated with the amount of blood ingested during feeding. Similarly, other hypothesized transmission routes such as coprophagy fail to explain the observed pattern of prevalence. Our results could have implications for the feasibility of late-acting control strategies that preferentially kill older insects.

scholarly journals Epidemiology and Control of Neosporosis and Neospora caninum

2007 ◽  
Vol 20 (2) ◽  
pp. 323-367 ◽  
Author(s):  
J. P. Dubey ◽  
G. Schares ◽  
L. M. Ortega-Mora
Keyword(s):  
Life Cycle ◽  
New Genus ◽  
Neospora Caninum ◽  
Protozoan Parasite ◽  
Zoonotic Potential ◽  
Human Tissues ◽  
New Genus And Species ◽  
Serious Disease ◽  
And Control

SUMMARY Neospora caninum is a protozoan parasite of animals. Until 1988, it was misidentified as Toxoplasma gondii. Since its first recognition in dogs in 1984 and the description of the new genus and species Neospora caninum in 1988, neosporosis has emerged as a serious disease of cattle and dogs worldwide. Abortions and neonatal mortality are a major problem in livestock operations, and neosporosis is a major cause of abortion in cattle. Although antibodies to N. caninum have been reported, the parasite has not been detected in human tissues. Thus, the zoonotic potential is uncertain. This review is focused mainly on the epidemiology and control of neosporosis in cattle, but worldwide seroprevalences of N. caninum in animals and humans are tabulated. The role of wildlife in the life cycle of N. caninum and strategies for the control of neosporosis in cattle are discussed.

Iron-modulated pseudocyst formation inTritrichomonas foetus

Parasitology ◽  
2016 ◽  
Vol 143 (8) ◽  
pp. 1034-1042 ◽  
Author(s):  
CÁSSIA CASTRO ◽  
RUBEM FIGUEIREDO SADOK MENNA-BARRETO ◽  
NILMA DE SOUZA FERNANDES ◽  
LEONARDO SABOIA-VAHIA ◽  
GEOVANE DIAS-LOPES ◽  
...  
Keyword(s):  
Life Cycle ◽  
Cell Biology ◽  
Proteolytic Enzymes ◽  
Essential Element ◽  
Protozoan Parasite ◽  
Transformation Process ◽  
Morphological Transformation ◽  
Iron Depletion ◽  
Transmission Electron

SUMMARYIron is an essential element for the survival of trichomonads during host–parasite interaction. The availability of this metal modulates several metabolic pathways of the parasites and regulates the expression of virulence factors such as adhesins and proteolytic enzymes. In this study, we investigated the effect of iron depletion on the morphology and life cycle ofTritrichomonas foetus. Scanning and transmission electron microscopy analyses revealed that depletion of iron from the culture medium (named TYM-DIP inducer medium) induces morphological transformation of typical pear-shaped trophozoites into spherical and non-motile pseudocysts. Remarkably, inoculation of pseudocysts into an iron-rich medium (standard TYM medium), or addition of FeSO4to a TYM-DIP inducer medium reverted the morphological transformation process and typical trophozoites were recovered. These results show that pseudocysts are viable forms of the parasite and highlight the role of iron as a modulator of the parasite phenotype. Although iron is required for the survival ofT. foetus, iron depletion does not cause a cellular collapse of pseudocysts, but instead induces phenotypic alterations, probably in order to allow the parasite to survive conditions of nutritional stress. Together, these findings support previous studies that suggest pseudocysts are a resistance form in the life cycle ofT. foetusand enable new approaches to understanding the multifactorial role of iron in the cell biology of this protozoan parasite.

Morphological characterization ofCryptosporidium parvumlife-cycle stages in anin vitromodel system

Parasitology ◽  
2009 ◽  
Vol 137 (1) ◽  
pp. 13-26 ◽  
Author(s):  
H. BOROWSKI ◽  
R. C. A. THOMPSON ◽  
T. ARMSTRONG ◽  
P. L. CLODE
Keyword(s):  
Electron Microscopy ◽  
Life Cycle ◽  
Protozoan Parasite ◽  
Morphological Characterization ◽  
Host Cells ◽  
Epithelial Cell Line ◽  
Life Cycle Stages ◽  
Complete Life Cycle ◽  
Behavioural Characteristics

SUMMARYCryptosporidium parvumis a zoonotic protozoan parasite that mainly affects the ileum of humans and livestock, with the potential to cause severe enteric disease. We describe the complete life cycle ofC. parvumin anin vitrosystem. Infected cultures of the human ileocecal epithelial cell line (HCT-8) were observed over time using electron microscopy. Additional data are presented on the morphology, development and behavioural characteristics of the different life-cycle stages as well as determining their time of occurrence after inoculation. Numerous stages ofC. parvumand their behaviour have been visualized and morphologically characterized for the first time using scanning electron microscopy. Further, parasite-host interactions and the effect ofC. parvumon host cells were also visualized. An improved understanding of the parasite's biology, proliferation and interactions with host cells will aid in the development of treatments for the disease.

scholarly journals RNase III Domain of KREPB9 and KREPB10 Association with Editosomes in Trypanosoma brucei

mSphere ◽  
2018 ◽  
Vol 3 (1) ◽  
Author(s):  
Jason Carnes ◽  
Suzanne M. McDermott ◽  
Kenneth Stuart
Keyword(s):  
Life Cycle ◽  
Trypanosoma Brucei ◽  
Protozoan Parasite ◽  
Tsetse Fly ◽  
Accessory Proteins ◽  
Rnase Iii ◽  
Parasite Life Cycle ◽  
Null Cell ◽  
Content Type

ABSTRACT Editosomes are the multiprotein complexes that catalyze the insertion and deletion of uridines to create translatable mRNAs in the mitochondria of kinetoplastids. Recognition and cleavage of a broad diversity of RNA substrates in vivo require three functionally distinct RNase III-type endonucleases, as well as five additional editosome proteins that contain noncatalytic RNase III domains. RNase III domains have recently been identified in the editosome accessory proteins KREPB9 and KREPB10, suggesting a role related to editing endonuclease function. In this report, we definitively show that KREPB9 and KREPB10 are not essential in either bloodstream-form parasites (BF) or procyclic-form parasites (PF) by creating null or conditional null cell lines. While preedited and edited transcripts are largely unaffected by the loss of KREPB9 in both PF and BF, loss of KREPB10 produces distinct responses in BF and PF. BF cells lacking KREPB10 also lack edited CYb, while PF cells have increased edited A6, RPS12, ND3, and COII after loss of KREPB10. We also demonstrate that mutation of the RNase III domain of either KREPB9 or KREPB10 results in decreased association with ~20S editosomes. Editosome interactions with KREPB9 and KREPB10 are therefore mediated by the noncatalytic RNase III domain, consistent with a role in endonuclease specialization in Trypanosoma brucei. IMPORTANCE Trypanosoma brucei is a protozoan parasite that causes African sleeping sickness. U insertion/deletion RNA editing in T. brucei generates mature mitochondrial mRNAs. Editing is essential for survival in mammalian hosts and tsetse fly vectors and is differentially regulated during the parasite life cycle. Three multiprotein “editosomes,” typified by exclusive RNase III endonucleases that act at distinct sites, catalyze editing. Here, we show that editosome accessory proteins KREPB9 and KREPB10 are not essential for mammalian blood- or insect-form parasite survival but have specific and differential effects on edited RNA abundance in different stages. We also characterize KREPB9 and KREPB10 noncatalytic RNase III domains and show they are essential for editosome association, potentially via dimerization with RNase III domains in other editosome proteins. This work enhances the understanding of distinct editosome and accessory protein functions, and thus differential editing, during the parasite life cycle and highlights the importance of RNase III domain interactions to editosome architecture.

Cardiac glycosides in Oncopeltus fasciatus (Dallas) (Hemiptera: Lygaeidae). I. The uptake and distribution of natural cardenolides in the body

1974 ◽  
Vol 52 (2) ◽  
pp. 283-290 ◽  
Author(s):  
S. S. Duffey ◽  
G. G. E. Scudder
Keyword(s):  
Body Weight ◽  
Life Cycle ◽  
Larval Development ◽  
Cardiac Glycosides ◽  
The Body ◽  
Oncopeltus Fasciatus ◽  
Chromatographic Behavior ◽  
Asclepias Syriaca ◽  
Metathoracic Gland

Oncopeltus fasciatus sequesters cardiac glycosides from seeds of Asclepias syriaca, and in the adult, concentrates these chemicals in the dorsolateral spaces of the thorax and abdomen; 60 to 95% of the cardenolide in such adults is stored in the dorsolateral complex. Very little cardenolide is detectable in the haemolymph of adults and larvae, but cardiac glycosides do occur in the adult ventral metathoracic gland secretions and in the larval middorsal abdominal gland fluid.O. fasciatus sequesters cardenolides throughout its life cycle. The complement of cardenolides acquired by feeding throughout the development is not related simply to increase in body weight. Unknown factors during late larval development are important in the accumulative uptake of the heart poisons.It is shown that O. fasciatus sequesters polar but not non-polar cardenolides into the dorsolateral complex in the adult. The accumulation of the polar cardenolides in both the larva and adult appears to be, at least in part, associated with the ability of the insect to metabolize heart poisons. The non-polar cardenolide digitoxin was converted to at least two more polar cardenolides. Adult O. fasciatus did not, however, metabolically alter the chromatographic behavior of ouabain that had been added to the food.

scholarly journals In Silico Investigation of Conserved miRNAs and Their Targets From the Expressed Sequence Tags in Neospora Caninum Genome

2021 ◽  
Vol 15 ◽  
pp. 117793222110467
Author(s):  
Moumita Das ◽  
Mahmudul Hasan ◽  
Sharmin Akter ◽  
Sawrab Roy ◽  
Binayok Sharma ◽  
...  
Keyword(s):  
Expressed Sequence Tags ◽  
Developmental Stages ◽  
Neospora Caninum ◽  
Protozoan Parasite ◽  
Study Finding ◽  
Mature Mirnas ◽  
Etiologic Agent ◽  
Homology Search ◽  
Specific Gene ◽  
Expressed Sequence

Neospora caninum is a protozoan parasite, the etiologic agent of Neosporosis—a common cause of abortion in cattle worldwide. Herd level prevalence of Neosporosis could be as high as 90%. However, there is no approved treatment and vaccines available for Neosporosis. MicroRNA (miRNA) based prophylaxis and therapeutics could be options for Neosporosis in cattle and other animals. The current study aimed to investigate the genome of Neospora caninum to identify and characterize the conserved miRNAs through Expressed Sequence Tags (ESTs) dependent homology search. A total of 1,041 mature miRNAs of reference organisms were employed against 336 non-redundant ESTs available in the genome of Neospora caninum. The study predicted one putative miRNA “nca-miR-9388-5p” of 19 nucleotides with MFEI value -1.51 kcal/mol and (A + U) content% 72.94% corresponding with its pre-miRNA. A comprehensive search for specific gene targets was performed and discovered 16 potential genes associated with different protozoal physiological functions. Significantly, the gene “Protein phosphatase” was found responsible for the virulence of Neospora caninum. The other genes were accounted for gene expression, vesicular transport, cell signaling, cell proliferation, DNA repair mechanism, and different developmental stages of the protozoon. Therefore, this study finding will provide pivotal information to future aspirants upon Bovine Neosporosis. It will also serve as the baseline information for further studies of the bioinformatics approach to identify other protozoal miRNAs.

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